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The Journal of Neuroscience, May 1, 1998, 18(9):3314-3326
Development of Bone Morphogenetic Protein Receptors in the
Nervous System and Possible Roles in Regulating trkC Expression
Damin
Zhang,
Mark F.
Mehler,
Qingbin
Song, and
John
A.
Kessler
Departments of Neurology and Neuroscience and the Rose F. Kennedy
Center for Research in Mental Retardation and Human Development, Albert
Einstein College of Medicine, Bronx, New York 10461
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ABSTRACT |
Characterization of bone morphogenetic protein receptor (BMPR)
expression during development is necessary for understanding the role
of these factors during neural maturation. In this study, in
situ hybridization analyses demonstrate that BMP-specific type I (BMPR-IA and BMPR-IB) and type II (BMPR-II) receptor mRNAs are expressed at significant levels in multiple regions of the CNS, cranial
ganglia, and peripheral sensory and autonomic ganglia during the
embryonic and neonatal periods. All three BMP receptor subunits are
expressed within periventricular generative zones. BMPR-IA is more
abundant than the other receptor subtypes, with widespread expression
in the brain, cranial ganglia, and peripheral ganglia. By contrast,
BMPR-IB mRNA displays significant expression within more restricted
regions, including the anterior olfactory nuclei. BMPR-II mRNA exhibits
peak expression within the cerebellar Purkinje cell layer and the
hippocampus, as well as within cranial ganglia. The distribution of BMP
receptors within large neurons in adult dorsal root ganglia suggested a
possible role in regulating expression of the neurotrophin receptor
trkC. This hypothesis was tested in explant cultures of embryonic day
15 (E15) and postnatal day 1 (P1) sympathetic superior cervical ganglia
(SCG). Treatment of the E15 or the P1 SCG with BMP-2 induced expression
of trkC mRNA and responsiveness of sympathetic neurons to NT3 as
measured by neurite outgrowth. The pattern of expression of BMP
receptors in embryonic brain suggests several potentially novel areas
for further developmental analysis and supports numerous recent studies that indicate that BMPs have a broad range of cellular functions during
neural development and in adult life.
Key words:
bone morphogenetic protein receptor; subventricular zone; cranial ganglia; dorsal root ganglia; gliogenesis; olfactory nuclei
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INTRODUCTION |
The bone morphogenetic proteins
(BMPs) are members of the transforming growth factor  (TGF-)
superfamily (for review, see Kingsley, 1994 ; Hogan, 1996). They are
involved in a number of cellular and molecular processes, including
bone formation (Urist et al., 1979; Wozney et al., 1988), mesoderm
induction (Dale et al., 1992), neurulation (Wilson and
Hemmati-Brivanlou, 1995), dorsoventral patterning of the neural tube
(Liem et al., 1995; Arkell and Beddington, 1997), apoptosis (Graham et
al., 1994), neural differentiation (for review, see Mehler et al.,
1997), and limb morphogenesis (Francis et al., 1994; Johnson et al., 1994; Kawakami et al., 1996). The BMPs are epidermal inducers that
inhibit neurulation, and anterior neurulation is orchestrated by
inhibition of BMP signaling (Wilson and Hemmati-Brivanlou, 1995; Tanabe
and Jessell, 1996; Hemmati-Brivanlou and Melton, 1997). Within the
neural tube they act as gradient morphogens to promote the
differentiation of dorsal cell types and participate through
cooperative signaling in specification of intermediate dorsoventral
cell types (Hogan, 1996; Holley et al., 1996; Piccolo et al., 1996;
Tanabe and Jessell, 1996; Zimmerman et al., 1996). They also regulate
segmentation of rhombomeres by inducing apoptosis of selected neural
crest-associated cellular populations (Graham et al., 1994, 1996). In
the peripheral nervous system (PNS) they act as instructive signals for
neuronal lineage commitment and regulate subsequent neuronal
differentiation (Varley et al., 1995; Shah et al., 1996; Reissmann et
al., 1996; Varley and Maxwell, 1996), whereas in the brain they promote
astroglial lineage commitment (Gross et al., 1996). They also
regulate neuronal survival and differentiation in both the brain and
the PNS (Lein et al., 1995, 1996; Mehler and Kessler, 1995).
BMPs exert their biological effects by binding to type I (BMPR-IA
and BMPR-IB) and type II (BMPR-II) receptor subunits (Koenig et al.,
1994; ten Dijke et al., 1994a; Nohno et al., 1995; Liu et al., 1995;
Rosenzweig et al., 1995) that are organized with minor modifications of
the prototypical TGF- subclass receptors (Massague, 1996). In
addition, different BMP subgroups may also signal through a second
class of receptors that are also activin receptors (ActRs). Here we
describe the regional and cellular localization of the three
BMP-specific receptor subunit mRNAs in the developing and adult mouse
nervous systems and the corresponding expression of two of the BMP
receptor subunit proteins. BMPR-IA mRNA exhibits the broadest range and
highest levels of expression in prenatal, postnatal, and adult mouse
brain, cranial ganglia, and peripheral sensory and sympathetic ganglia.
In addition, expression of BMPR-IB and BMPR-II mRNAs is prominent
during the early postnatal period in several regions of mouse brain and
in cranial ganglia. The pattern of expression suggested several
potential functions for the BMPs, including a possible role in
regulating expression of the neurotrophin receptor trkC. As
predicted, BMP-2 treatment of explants of the superior cervical
ganglion was found to induce trkC expression as well as neurotrophin 3 (NT3) responsiveness. More generally, characterization of the
development and distribution of BMP receptor transcripts in developing
brain and peripheral ganglia provide the basis for predicting other
potential roles for the ligands in neural development.
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MATERIALS AND METHODS |
Tissue preparation. CD1 mouse embryos were fixed in
4% paraformaldehyde in PBS at 4°C overnight. After fixation, tissue
was exposed to 30% sucrose overnight at 4°C and then frozen for
cryostat sectioning. The postnatal mouse brain was directly chilled in liquid nitrogen for cryostat sectioning without prefixation. Sections (12-16 µm) were cut, post-fixed in 4% paraformaldehyde in PBS for
15 min, rinsed twice in PBS, dehydrated through a graded series of
ethanol washes, and stored at  70°C before use.
Preparation of cRNA probes. The probe used to survey the
expression of BMPR-IA was a 262 bp fragment corresponding to mouse BMPR-IA cDNA sequence, 110-372 bp (Dewulf et al., 1995). The BMPR-IB riboprobe was transcribed from a 486 bp fragment corresponding to mouse
BMPR-IB cDNA sequence, 359-845 bp (ten Dijke et al., 1994b). The
BMPR-II riboprobe was transcribed from a 348 bp fragment corresponding
to human BMPR-II cDNA sequence, 484-832 bp (Nohno et al., 1995).
Reverse transcription-PCR was subsequently performed. The fragments
were inserted into the corresponding plasmids: pGEM-78f(+/) plasmid
was used for type I receptors, and AT plasmid was used for the type II
receptor. The trkC probe was generated from a 676 bp fragment that
includes part of the kinase domain. Antisense and sense riboprobes were
transcribed with a Promega (Madison, WI) kit following the
manufacturer's instructions.
In situ hybridization. In situ hybridization was
performed by post-fixing sections in 0.1 M sodium
phosphate-buffered 4% paraformaldehyde, pH 7.4, for 30 min, rinsing in
PBS for 1 min, rinsing in 2× SSC for 1 min, acetylation with 0.5%
acetic anhydride in 0.1 M triethanolamine, pH 8.0, for 10 min, rinsing again in 2× SSC and then in PBS, and finally dehydrating
in a graded series of ethanol washes. The slides were prehybridized in
2× SSC and 50% formamide at 50°C for 2 hr and hybridized using
hybridization buffer containing 2 × 104
cpm/µl cRNA probe (hybridization buffer, 0.75 M NaCl,
50% formamide, 1× Denhardt's solution, 10% dextran sulfate, 30 mM DTT, 10 mM Tris-HCl, pH 7.5, 1 mM EDTA, 100 ug/ml salmon sperm DNA, and 0.5 mg/ml yeast
tRNA) at 50°C for 16 hr. Slides were then washed twice in 2× SSC for
2 min; in 2× SSC, 50% formamide, and 0.1% -mercaptoethanol (BME)
at 50°C for 1 hr; in 20 µg/ml RNase A at 37°C for 30 min; in 0.5 M NaCl and 10 mM Tris-HCl, pH 8.0; in 2× SSC,
50% formamide, and 0.1% BME at 58°C for 30 min; and in 0.1× SSC
and 0.1% BME at 63°C for 30 min, with final dehydration. The
sections were then exposed to x-ray film for 4 or 5 d to obtain
autoradiograms, dipped in emulsion, and exposed for 6-8 weeks, with
cresyl violet used as counterstain. In cases in which in
situ hybridization was combined with immunocytochemistry, slides
were processed for immunocytochemical analysis before radioautographic
detection of 35S. Use of the sense riboprobe confirmed the
specificity of labeling.
Ribonuclease protection assay. Antisense riboprobes for
BMP-IA, BMP-IB, BMPR-II, and trkC were generated in 25 µl of buffer containing 200 mM Tris-Cl, pH 7.5, 30 mM MgCl,
10 mM spermidine, 50 mM NaCl, 0.4 mM ATP, 0.4 mM GTP, 0.4 mM UTP, 10 ml of [ -32P]CTP, 800 Ci/mmol, 20 mM DTT,
20 U of ribonuclease inhibitor, 0.5 mg of template, and 20 U of T7 or
SP6 RNA polymerase. The mixture was incubated at 40°C for SP6 or at
37°C for T7 (60 min). Probes were purified using gel filtration. mRNA
from mouse brains (10 µg) was hybridized with 5 × 105 cpm of BMPR RNA probes at 60°C overnight. The
mixture was digested with 40 µg/ml ribonuclease A and 2 µg/ml
ribonuclease T1. Hybridized RNAs were run on 5% polyacrylamide gels
and visualized by autoradiography. mRNA from cultured ganglia (5 µg)
for analysis of trkC and actin was treated similarly, except that the
concentration of ribonuclease T1 was 2.5 µg/ml.
Immunoblotting. Cellular suspensions of mouse brains
(Charles River Laboratories, Wilmington, MA) in lysis buffer (1%
Triton X-100, 0.1% SDS, 1% sodium deoxycholate, 1 mM
iodoacetamide, 0.2 mg/ml aprotinin, 1 mM
phenylmethylsulfonyl fluoride, 0.01 M Tris-Cl, pH 8.0, 0.14 M NaCl, and 0.025% NaN3) were shaken
for 1 hr at 4°C. Insoluble debris was removed by centrifugation at
10,000 rpm for 10 min. Protein mixtures (~5 µg) were loaded on 8%
SDS-PAGE gels and transferred to nitrocellulose membranes. Membranes
were probed with affinity-purified BMPR-IA, BMPR-IB, and BMPR-II
antibodies (a generous gift from Dr. K. Miyazono, The Cancer Institute,
Tokyo, Japan). Blots were developed using the ECL reagent (Amersham, Arlington Heights, IL).
Immunocytochemistry. After the slides were processed through
in situ hybridization protocols, they were
immunocytochemically stained using an avidin-biotin-peroxidase
technique (Vectastain Elite mouse IgG ABC kit; Vector Laboratories,
Burlingame, CA). Neurons were identified by the presence of signal to
anti-NF68 antibodies (1:200 dilution; Sigma, St. Louis, MO).
Oligodendrocytes were identified with anti-CNPase antibodies (1:1000
dilution; Sternberger Monoclonals Inc.). Astrocytes were identified
with anti-glial fibrillary acidic protein (GFAP) antibodies (1:400 dilution; Sigma).
Culture of sympathetic ganglia. Superior cervical ganglia
were dissected from E15 and neonatal mice and were cultured as
described previously (Kessler et al., 1981) on a collagen substratum in serum-free medium (Neurobasal Plus B27 supplements; Life Technologies, Gaithersburg, MD). The E15 ganglia were cultured in the absence of NGF,
whereas neonatal ganglia were all treated with NGF (10 ng/ml).
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RESULTS |
To determine the spatial and temporal patterns of expression of
BMP receptors, mouse brains were processed for in situ
hybridization using 35S-labeled BMP receptor riboprobes.
The specificity of the hybridization signals was determined using two
separate techniques. First, 35S-labeled sense riboprobes
produced only background levels of hybridization (see Fig.
7g,h). Second, in RNase protection assays each cRNA probe
hybridized specifically with a single poly(A+) RNA
transcript isolated from mouse brain (Fig.
1A). Furthermore, immunoblotting with affinity-purified polyclonal antibodies to the BMP
receptor subunits demonstrated developmental regulation of protein
expression (Fig. 1B) that correlated with the results of the in situ studies.
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Figure 1.
A, Expression of BMPR-IA,
BMPR-IB, and BMPR-II mRNAs in neonatal mouse
whole brain samples as detected by RNase protection assays.
B, Western blot analyses of BMPR-IA and BMPR-II proteins
in embryonic (E9, E13, E16), postnatal
(P1, P7, P12), and adult (Ad) whole
brains.
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Western blot analysis of BMPR-IA in whole brain from embryonic day 9 (E9) to adult revealed an absence of detectable expression at embryonic
day 9, peak protein levels from embryonic day 13 to postnatal day 7 (P7), and reduced expression at P12 and in the adult (Fig.
1B). A similar pattern of expression was observed for
the BMPR-II protein, except that abundance at P12 and in adult brain
was much lower (Fig. 1B). Western blot analysis of
BMPR-1B expression revealed multiple bands, precluding detailed
developmental analysis (data not shown).
Differential expression of BMP-IA, BMPR-IB, and BMPR-II mRNAs in
the developing nervous system
All three receptor subunits are expressed at E12, and the patterns
of expression do not change substantially between E12 and E15. During
this period, BMPR-IA mRNA is most abundantly expressed in the
periventricular ventricular zone (VZ) and in the trigeminal ganglion
(Fig. 2), whereas BMPR-IB mRNA is
expressed within the anterior portion of the periventricular VZ (e.g.,
olfactory neuroepithelium and rhinencephalic neuroepithelium) and the
olfactory primordia (Fig. 2). By contrast, BMPR-II mRNA at these early
developmental stages is expressed within the entire extent of the VZ
(Fig. 2). These results suggest that all three BMP receptor subunits
may be involved in neural development at early embryonic stages.
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Figure 2.
Expression of BMP receptor subunit mRNAs as
detected by in situ hybridization and radioautography in
sagittal sections of E15 mouse head. BMPR-IA mRNA is
expressed by the VZ (arrow) and the trigeminal ganglion
(small arrow) in the nervous system. The expression of
BMPR-IB mRNA is restricted to the frontal region of the
VZ (arrow) and the olfactory epithelium
(arrowhead). BMPR-II mRNA is also
expressed in the VZ (arrow) and the trigeminal
ganglion (small arrow). Scale bar, 1 mm.
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At E16, BMPR-IA mRNA is most intensively expressed by cranial and
peripheral ganglia, including the trigeminal ganglion, nodose ganglion,
sympathetic ganglia, and dorsal root ganglia (DRG) (Figs. 3, 4). In
brain, BMPR-IA is expressed within multiple structures, including the
striatal neuroepithelium, the mitral cell layer, and the facial nuclei.
The thalamus exhibits less abundant expression of BMPR-IA mRNA, and
there are multiple other areas of low-level expression. By contrast to
the wide expression of BMPR-IA, BMPR-IB mRNA at this stage is limited
to the frontal region of the neuroepithelium and the olfactory
epithelium, analogous to the pattern at E12 and E15 (Fig. 3). In the
PNS, intensive labeling is seen within the nodose ganglion (data not
shown) with persistence through E19. However, BMPR-IB is not detectable
in trigeminal ganglion or DRG at these stages. BMPR-II mRNA is most
abundant within the neuroepithelium surrounding the ventricle at E16
(Fig. 3). The mRNA is also expressed in the trigeminal and nodose
ganglia (data not shown).
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Figure 3.
Expression of BMP receptor subunit mRNAs in the
E16 mouse head. BMPR-IA mRNA is expressed at high levels
in the trigeminal ganglion (arrow), nodose ganglion
(small arrow), and DRG (arrowhead). It is
also expressed by the striatum and the contiguous periventricular
generative zone (small arrows). Relatively weak and
diffuse expression is found in the thalamus (TH).
Expression of BMPR-IB mRNA is limited to the frontal
region of the periventricular generative zone (arrow)
and to the olfactory epithelium (arrowhead).
BMPR-II mRNA is also expressed by the periventricular
generative zone (arrow). Scale bar, 1 mm.
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Figure 4.
Expression of BMPR-IA mRNA in DRG during
development. BMPR-IA mRNA is expressed homogeneously in DRG
(cervical region) in the E16 mouse
embryo. The abundant expression persists in P0 DRG
(arrow, cervical region) and in Adult DRG
(shown at higher magnification). Notice that in Adult,
BMPR-IA mRNA is expressed primarily by large neuronal cells. Scale
bars: E16 and P0, 300 µm;
Adult, 100 µm.
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The intensive expression of BMPR-IA mRNA in DRG persists from E15
through later stages of development and into the adult (Fig. 4). In
early embryos (E15), the mRNA is expressed in a homogeneous pattern
throughout the DRG. By contrast, in later developmental stages and in
the adult BMPR-IA mRNA expression is restricted to a subpopulation of
larger, peripherally located neurons (Fig. 4). In the sympathetic
superior cervical ganglion, however, there is a homogeneous pattern of
expression throughout development and into the adult (see Fig.
11D).
At E17-E18, BMPR-IA mRNA expression increases in the thalamus (Fig.
5). The mRNA is also found in
hippocampus, periventricular subventricular zone (SVZ), superficial
regions of the cortical plate, and the mitral cell layer. By contrast,
BMPR-IB and BMPR-II are largely limited to the SVZ in brain at E17-E19
(data not shown). In addition, at E19, BMPR-IB mRNA is intensively
expressed on the anterior olfactory nuclei (data not shown).
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Figure 5.
Expression of BMPR-IA mRNA in the
E17 and E18 mouse head. BMPR-IA mRNA is
expressed abundantly by the trigeminal ganglion (arrow)
and nodose ganglion (arrowhead) at E17.
It is also expressed homogeneously in the thalamus, mitral cell layer
(small arrowheads), striatal generative zone
(small arrows), cortical plate, and the primordium of
the teeth. Note that in the high-power image of E18
mouse brain, BMPR-IA mRNA is expressed in the thalamus, hippocampus,
VZ/SVZ (arrow), and cortical plate
(arrowhead). TH, Thalamus;
HI, hippocampus; PT, primordium of the
teeth. Scale bars: E17, 1 mm; E18, 200 µm.
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At P2, BMPR-IA mRNA is intensively expressed within the SVZ, frontal
neocortex, striatum, piriform cortex, and taeniatecta (data not shown).
By contrast, BMPR-IB mRNA is intensively expressed only in the anterior
olfactory nuclei (Fig. 6) and in the SVZ (data not shown). The abundant expression of BMPR-IB mRNA in the olfactory epithelium correlates well with the early onset and persistent expression of BMPR-IB mRNA in anterior olfactory nuclei, suggesting that BMP-IB-mediated signaling may play an important role on
the development and functioning of the olfactory system. BMPR-II mRNA
is intensively expressed within the periventricular region (Fig.
6).
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Figure 6.
Expression of BMPR mRNAs within frontal cortical
regions of the P2 mouse brain. BMPR-IB mRNA expression
is limited to the anterior olfactory nuclei (arrow),
whereas BMPR-II is expressed at high levels within the
region of the SVZ (arrow). Scale bar, 500 µm.
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During P0-P14, BMPR-IA mRNA is broadly expressed in the mouse brain.
In the neocortex, the most prominent expression is in the supragranular
portion of the cortical plate. Expression within the thalamus at P4 is
uniform (Fig. 7a), with no
preferential labeling within specific thalamic nuclei; more specific
labeling of thalamic nuclei begins at P6. BMPR-IA mRNA labeling is weak in the Purkinje cell layer of the cerebellum during the neonatal period
(Fig. 7a). Intense and specific expression is evident over the ependymal layer of the SVZ (Fig. 7a); such ependymal
expression is not found in either P14 or the adult brain (Figs.
7b, 8). Although the dentate
gyrus is not labeled, the other regions of the hippocampus display
intensive labeling that persists at later developmental stages. Several
other regional structures also exhibit patterns of BMPR-IA mRNA
expression similar to those seen in P14 and adult brains. For instance,
the pontine gray nuclei and the superior and inferior olivary nuclei
express significant levels of BMPR-IA mRNA. The mitral cell layer of
the olfactory bulb also displays intensive expression of BMPR-IA mRNA
as seen in P14 and adult brain. By P14, the pattern of BMPR-IA
expression is very similar to the one seen in adult brain, although
labeling appears more diffuse. Strong expression is seen within the
hippocampus, neocortex, mitral cell layer of the olfactory bulb,
cerebellar Purkinje cell layer, and in nuclei of the thalamus and
brainstem (Fig. 7b). In the neocortex, the labeling is
differentially distributed over the various cell layers; the most
intense labeling is seen over layers III, IV, and V, similar to the
overall pattern seen in the adult cortex (Fig. 7b). BMPR-IA
mRNA is also expressed within specific nuclei in the thalamus, such as
the anterior dorsal, reticular, and ventral posterior thalamic nuclei.
Unlike the adult brain, the choroid plexus of P14 mouse brain does not
express BMPR-IA mRNA.
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Figure 7.
Expression of BMP receptor mRNAs at postnatal days
4 and 14. BMPR-IA mRNA is widely distributed in the
P4 (a) and P14
(b) mouse brains. Notice that the expression
pattern within the thalamus changes from a homogeneous pattern at
P4 (arrow) to expression within specific
nuclei at P14 (arrow). The expression of
BMPR-IB is restricted to the anterior olfactory nuclei
at P4 (c) and P14
(d). The expression of BMPR-II is
seen within the hippocampus, cerebellum (arrow), and
mitral cell layer of the olfactory bulb at P4
(e) and P14
(f). Notice that all three receptors are
specifically expressed within the ependymal layer of the SVZ at
P4 (a, c, e; arrowhead), but not at older
stages, such as P14 (b, d, f; arrowhead).
BMPR-II is expressed within the cerebellum in the postnatal period at
higher levels than BMPR-IA. g, h, Control
sections hybridized with BMPR-IA sense probe showing background
labeling. Scale bars: a, c, e, g, 2 mm; b, d, f,
h, 2 mm.
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Figure 8.
The expression of BMP receptor subunit mRNAs as
detected by in situ hybridization and radioautography in
sagittal sections of adult mouse brain. BMPR-IA mRNA is
widely expressed throughout the brain. Intense labeling is seen within
the cerebral cortex (CX), hippocampus
(small arrow), Purkinje cell layer of the cerebellum
(arrow), brainstem, thalamic nuclei (small arrow
heads), mitral cell layer of olfactory bulb
(MCL), and choroid plexus of the lateral ventricle
(arrowhead). The expression of BMPR-IB
mRNA is restricted to the anterior olfactory nuclei
(arrow). BMPR-II mRNA is expressed at
modest levels within the hippocampus (arrow), choroid
plexus (arrowhead), Purkinje cell layer of the
cerebellum (small arrow), and the mitral cell layer of
the olfactory bulb. Scale bar, 2 mm.
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The expression of BMPR-IB mRNA within the anterior olfactory nuclei
(AON) persists from the neonatal period to the adult (Figs. 7c,d, 8). However, the intensity of expression in the AON
changes during development, with maximal expression at P14. These
experimental observations suggest that BMPR-IB signaling may play a
particularly prominent role in the maturation of neurons in the
anterior olfactory nuclei. Furthermore, during the neonatal period,
BMPR-IB mRNA is also discretely expressed in the ependymal layer of the
SVZ (Fig. 7c).
At P4 and also at earlier stages, intense BMPR-II expression is found
over the ependymal layer of the SVZ (Fig. 7e). The primordia of the cerebellar Purkinje cell layer and the facial nuclei in the
brainstem also express BMPR-II at P0-P4. In the hippocampus, high
levels of BMPR-II mRNA are found at P14; by contrast, expression is
less abundant both during the neonatal period and in the adult (Figs.
7e,f, 8). Similarly, in comparison with the neonatal and adult period, the expression of BMPR-II in the cerebellar Purkinje cell
layer is most apparent at P14. These experimental findings suggest that
BMPR-II-mediated signaling may play an important role in the
development of the cerebellum during the postnatal period.
Differential expression of BMPR-IA, BMPR-IB, and BMPR-II mRNAs in
adult mouse brain
All three BMP receptor subunit mRNAs are expressed in adult brain,
but each exhibits distinct patterns of regional expression (Fig. 8).
BMPR-IA mRNA is more abundant, with a broader pattern of distribution
than either BMPR-IB or BMPR-II mRNA. The patterns of expression and
intensity of BMPR-IA mRNA signals vary widely within specific gray
matter regions. No BMPR-IA mRNA expression is detectable in white
matter structures, including the corpus callosum, internal capsule,
posterior or anterior commissures, or in the cerebral peduncles. In the
telencephalon, the expression of BMPR-IA mRNA is greatest in the mitral
cell layer of the olfactory bulb (Fig. 8). In more caudal regions,
hybridization is distributed across the cellular layers of the anterior
olfactory nucleus, the olfactory tubercle, and the piriform cortex. In
the neocortex, labeled cells are distributed across all neuronal
layers, with the greatest intensity in layers IV and V (Figs. 8,
9a,b). The autoradiographic
grains are primarily localized to large neuronal perikarya, but the
cortical neuropil display diffuse labeling that exceeds the general
autoradiographic background. In the basal forebrain, weak and diffuse
labeling is found in association with the caudate and putamen without
clear association with specific cells or distinct cytoarchitectonic
fields of these nuclei. Within hippocampus, the most intense labeling
occurs in the CA regions and in the dentate gyrus (Figs. 8,
9a). In the diencephalon at the level of the dorsal
thalamus, a distinct subset of nuclei are labeled by the BMPR-IA
riboprobe, including the lateral dorsal, medial dorsal, paraventricular
thalamic nuclei, and the zona incerta (Figs. 8, 9a). The
amygdaloid nuclei and dorsal medial hypothalamic nucleus also
specifically express BMPR-IA mRNA (data not shown). BMPR-IA is broadly
expressed within the mesencephalon, metencephalon and myelencephalon.
The highest levels of regional expression occur in the cerebellar
Purkinje cell layer and within nuclei of the brainstem. In the pons,
BMPR-IA mRNA is expressed at significant levels in several pontine and
pontine reticular nuclei (Fig. 8). In the cerebellum, strong expression
of BMPR-IA mRNA is found in the dentate, interpositus, and fastigial
cerebellar nuclei (data not shown). A sharp band of hybridization
clearly distinguishes the Purkinje cell layer from the surrounding
granule cell and molecular layers (Fig. 8). The ependymal lining of the
lateral ventricle was not labeled in the adult brain. However, it
should be noted that hybridization signal is evident in the choroid
plexus of the lateral ventricle (Fig. 8) and the fourth ventricle.
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Figure 9.
Expression of BMPR-IA mRNA in the adult mouse
brain. a, Coronal section of adult brain through the
region of the thalamus. Notice that BMPR-IA mRNA is expressed in medial
dorsal and paraventricular thalamic nuclei (arrow),
lateral dorsal nuclei (arrowhead), zona incerta
(small arrows), and dorsal medial hypothalamic nucleus
(small arrowhead). b, c,
Cerebral cortical regions at high magnification. Notice in
c that BMPR-IA mRNA is colocalized with immunostaining
for neurofilament, indicative of expression in neurons. Scale bars:
a, 1 mm; b, 200 µm;
c, 100 µm.
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Although almost all transcript labeling appeared to be concentrated on
large neuronal cell bodies within discrete nuclei and cortical cell
layers, it is possible that astrocytes or oligodendrocytes in certain
brain regions also expressed BMPR-IA mRNA. Immunohistochemistry combined with in situ hybridization was applied to address
this issue using a neurofilament antibody (NF68) as a marker for
neurons, CNPase as a marker for oligodendrocytes, and GFAP as a marker for astrocytes. Colocalization was detected only with the neurofilament antibody (Fig. 9c), suggesting that the expression of
BMPR-IA is largely confined to neuronal subpopulations.
The expression pattern of BMPR-IB mRNA in adult brain is much more
limited. The anterior olfactory nucleus is the sole regional structure
that displays strong BMPR-IB transcript expression (Fig. 8). BMPR-II is
expressed more abundantly with a wider pattern of distribution than
BMPR-IB. Transcripts for BMPR-II are seen in the hippocampus (all
regions), the choroid plexus of the lateral and IV ventricles, the
cerebellar Purkinje cell layer, and the mitral cell layer of the
olfactory bulb (Fig. 8). However, levels of expression of BMPR-II are
of relatively lower intensity. BMPR-II expression is conspicuously
absent in the areas expressing BMPR-IB, suggesting that there may be a
second type II subunit associated with the BMPR-IB subunit. These
distinct patterns of transcript expression indicate that the three
BMP-specific receptor subunits reside in specific regional cellular
subpopulations, and may thus participate in mediating distinct
functional roles for individual BMPs in the normal adult brain.
Induction of trkC in sympathetic ganglia by BMP-2
The pattern of expression of BMP receptors in large DRG neurons
(Fig. 4) suggested that these receptors might be associated with
expression of the neurotrophin receptor trkC that is similarly distributed in DRG (Mu et al., 1993). Because previous studies have
suggested that the BMPs are involved in regulating growth factor
responses in neural crest derivatives (Song et al., 1998), we
hypothesized that BMPs induce trkC expression. Sensory ganglia normally
express trkC throughout development and also appear to preferentially
express BMP receptors in neurons already expressing trkC. Therefore,
the sympathetic superior cervical ganglion (SCG), a neural crest
derivative that does not normally express high levels of trkC after E15
(Fagan et al., 1996) and that expresses BMP receptors relatively
uniformly throughout the ganglion (see Fig. 11D), was
used to test this hypothesis. Ganglia were treated with BMP-2 and
examined for levels of trkC mRNA (Fig.
10). E15 sympathetic neurons survive in
culture without exogenous growth factors (Coughlin and Collins, 1985).
Explants of E15 SCG were therefore cultured with no added factors
or with BMP-2 and were subsequently examined for levels of trkC
mRNA and actin mRNA (Fig. 10). Untreated ganglia contained minimally
detectable levels of trkC. By contrast, treatment with BMP-2 increased
levels of trkC mRNA >40-fold. The effects of BMP-2 were also examined
in explants of the neonatal SCG (Fig. 10). Because sympathetic neurons
at this age are dependent on NGF for survival, all neonatal cultures
were treated with this factor. Ganglia treated with NGF alone contained very low levels of trkC mRNA. By comparison, treatment with BMP-2 increased levels of trkC mRNA >25-fold. Thus BMP-2 treatment induced expression of trkC at both developmental stages. Further,
BMPR-1A expression persisted even in the adult SCG (Fig.
11D), suggesting ongoing responsiveness to BMP-2 throughout development.
View larger version (38K):
[in this window]
[in a new window]
|
Figure 10.
Regulation of trkC expression by BMP-2.
A, Explants of E15 and of neonatal superior cervical
ganglia were cultured in the absence or presence of BMP-2 (10 ng/ml)
for 48 hr and were examined by nuclease protection assay for levels of
trkC mRNA. Levels of actin mRNA were simultaneously determined to
assure equivalent loading of RNA in each lane. Note that
BMP-2 treatment increased levels of trkC
mRNA in both E15 and neonatal explants.
B, Levels of trkC mRNA in the nuclease
protection assays were quantitated densitometrically and normalized to
levels of actin mRNA in each sample. The level in
E15 control ganglia was assigned a value of 1, and other
samples are normalized to this control value. *Differs from respective
control value at p < 0.01; n = 4.
|
|
View larger version (113K):
[in this window]
[in a new window]
|
Figure 11.
Effects of BMP-2 treatment on neurite extension
by sympathetic ganglia. E15 sympathetic ganglia were cultured in the
absence of added factors or with NT3 (20 ng/ml; A),
BMP-2 (20 ng/ml; B), or NT3 and BMP-2
(C). Like untreated ganglia (data not shown),
neurite outgrowth was minimal with NT3 alone or BMP-2 alone but was
much more extensive after combined treatment with NT3 and BMP-2.
D, In situ hybridization analysis of an
adult SCG for BMPR-1A mRNA. Note that the mRNA is still expressed
abundantly even in the adult. Scale bars in A-C, 130 µm.
|
|
Induction of NT3 responsiveness by BMP-2
To determine whether the induction of trkC was associated with
increased NT3 responsiveness, E15 SCG explants were treated with NT3
either with or without BMP-2 cotreatment and were examined for neurite
outgrowth (Fig. 11). Control (untreated) ganglia extended virtually no
neurites during the first 48 hr in culture (data not shown). Similarly,
ganglia treated with BMP-2 alone (Fig. 11A) or NT3
alone (Fig. 11B) extended few neurites. However,
neurite outgrowth from ganglia treated with both BMP-2 and NT3 was
extensive (Fig. 11C), indicating that expression of trkC
after BMP-2 treatment is associated with a gain of NT3
responsiveness.
| |
DISCUSSION |
BMP subclass ligands, which subserve multiple functions during the
development of the nervous system (for review, see Mehler et al.,
1997), exert their biological functions through binding to both
subclass-specific (BMPRs) and subclass-selective (ActR) subunits.
Therefore, localization of BMP receptors in specific regional cellular
populations will help define the role of these TGF- subclass factors
in the developing and mature nervous system.
BMPs may play a role in suppression of neurogenesis and neuronal
lineage commitment in the brain
The BMPs and their receptors are expressed in very early stages of
Xenopus development (Hemmati-Brivanlou and Thomsen, 1995), in which they actively suppress neurogenesis (Wilson and
Hemmati-Brivanlou, 1995; Tanabe and Jessell, 1996). In the avian
rhombencephalon, this early expression is associated with apoptosis of
selected populations of cells, leading to segmentation (Graham et al., 1994). We found that the receptors for the BMPs are expressed within
CNS neuroepithelium at E12, and expression persisted largely unchanged
until E15 (Fig. 2). This period precedes gliogenesis but is temporally
associated with the generation of neurons. The precise role of the BMPs
at this time remains to be determined. However, a preponderance of
evidence suggests that these factors suppress neurogenesis and neuronal
lineage commitment in the brain (Wilson and Hemmati-Brivanlou, 1995;
Gross et al., 1996; Tanabe and Jessell, 1996). By contrast, the BMPs
appear to induce neuronal lineage commitment of neural crest progenitor
cells (Shah et al., 1996), suggesting that their actions may be
population-specific.
BMPs may also play an important role in the differentiation and
survival of neurons within cranial ganglia and the DRG
BMPR-IA mRNA is intensively and homogeneously expressed in the DRG
from E15 to P0. By contrast, in adult DRG BMPR-IA mRNA expression is
restricted to large-sized neurons located in the periphery of the
ganglion. Transcripts for all three receptor subunits are expressed in
nodose ganglia, and BMPR-IA and BMPR-II mRNAs are also expressed in
trigeminal and sympathetic ganglia. The expression of BMP receptor
subunit transcripts in cranial ganglia, DRG, and sympathetic ganglia
correlates with observations that BMPs regulate the cellular maturation
of neural crest-derived progenitor cells, (Graham et al., 1994; Varley
et al., 1995; Reissmann et al., 1996; Shah et al., 1996; Varley and
Maxwell, 1996) as well as later stages of development of certain neural
crest derivatives (Fann and Patterson, 1994; Lein et al., 1995, 1996).
The precise function of the BMPs in neuronal development is unclear.
BMP treatment of sympathetic progenitor cells enhances
catecholaminergic expression (Varley et al., 1995; Reissmann et al.,
1996; Varley and Maxwell, 1996), and treatment of more mature
sympathetic neurons in culture alters their peptide neurotransmitter
phenotype and fosters the outgrowth of dendrites in preference to axons
(Fann and Patterson, 1994; Lein et al., 1995, 1996).
The decapentaplegic subgroup of the BMPs (BMP-2 and BMP-4), which
interact with the BMP-specific receptor subunits examined in this study
(Koenig et al., 1994; Mishina et al., 1995), may also participate in
the induction of growth factor dependence of neural crest derivatives
and may promote apoptosis in cells not exposed to other factors (Song
et al., 1998). Treatment of MAH cells, an immortalized sympathoadrenal
progenitor cell line (Birren and Anderson, 1990), with BMP-2 or
BMP-4 induces a requirement for other growth factors to prevent
apoptosis (Song et al., 1998), and BMP-4 induces apoptosis of
rhombencephalic neural crest (Graham et al., 1994). In this study we
found that BMP-2 treatment of sympathetic neurons increases expression
of trkC and NT3 responsiveness. E15 sympathetic neurons are already
responsive to NGF, and they spontaneously become dependent on
neurotrophins after 48 hr in culture even without BMP treatment.
Therefore, no conclusions can be drawn regarding effects of BMP-4 in
inducing neurotrophin dependence as well responsiveness to NT3.
Nevertheless these observations support the thesis that the BMPs
participate in the induction of growth factor responses, particularly
to the neurotrophins. The early expression of BMP receptors by
neurotrophin-responsive cell populations (DRG, sympathetic ganglia,
cranial ganglia, and striatal neurons) is consistent with such a
postulated role.
Expression of all three BMP receptor subunits within the
subventricular zones during the late embryonic and neonatal periods
suggests that the BMPs may play diverse cellular roles, including
the regulation of gliogenesis
All three BMP receptors, BMPR-IA, BMPR-IB, and BMPR-II, are
specifically expressed within the subventricular zones with maximal expression from E19 to P4, coincident with the period of cortical gliogenesis. These results support the idea that BMPs play an important role in the promotion of astroglial lineage commitment from
SVZ progenitor cells (Gross et al., 1996). BMPs induce the transient
expression of GFAP-immunoreactive astrocytes from cultured mouse
embryonic stem cells (D'Alessandro et al., 1994). They also induce the
stable elaboration of astrocytes from cultured murine SZV progenitor
cells with concurrent suppression of oligodendroglial and
neuronal differentiation (Gross et al., 1996). Furthermore, BMP ligands
are expressed in the subventricular region and in developing brain
during the late embryonic and neonatal periods (Mehler and Kessler,
1995; Soderstrom et al., 1996). These cumulative observations
suggest that the BMPs may participate in the elaboration of the
astrocytic lineage. It is interesting that all three BMP receptor
subunits were expressed in the SVZ and that members of several
different BMP subgroups promote astrocytic differentiation (Gross et
al., 1996), suggesting that activation of several different BMP
receptor subtypes may foster gliogenesis. Similarly, these receptors
all appear to participate in the suppression of neuronal lineage
commitment in the brain (Gross et al., 1996).
Widespread expression of BMPR-IA mRNA in adult brain suggests that
specific BMP ligands may have important roles in regulating neural
function in the adult
Transcripts for the BMPR-IA receptor subunit are broadly expressed
in the adult brain. Expression appears to be restricted largely to
neurons, with particularly robust patterns of expression in
subpopulations within the neocortex and brainstem nuclei. These observations suggest that specific BMP ligands may exert multiple functions in the adult brain. BMPR-IA may be involved in several different signal transduction pathways that may be partially determined by different complements of receptor subunits in distinct cell types.
It has been reported that BMP-2 and BMP-4 bind specifically to BMPR-IA
(Koenig et al., 1994; Mishina et al., 1995). However, the distinct
roles of BMP-2 and BMP-4 in adult brain have yet to be documented;
homozygous null mutants for BMPR-IA, BMPR-2, and BMP-4 are embryonic
lethal (Mishina et al., 1995; Winnier et al., 1995; Zhang and Bradley,
1996). Our results differ from the observations of Soderstrom et
al. (1996), who reported that transcripts for BMPR-IA and BMPR-IB are
not expressed in the adult brain. This discrepancy may be attributable
to the type of antisense probe used for the different in
situ hybridization studies. However, the nuclease protection
assays and Western blot analyses (Fig. 1) leave little doubt that these
receptors are actually expressed in mouse brain. Although the functions
of the BMPs in the adult remain unclear, increasing evidence suggests
that the BMPs promote dendritic elongation and branching (Lein et al.,
1995, 1996), raising the possibility that these factors regulate
dendritic growth in the mature brain.
The specific expression of BMPR-IB mRNA in anterior
olfactory nuclei suggests a role for some BMP ligands in the regulation
of the olfactory system
In contrast to the broad expression of BMPR-IA mRNA in brain,
BMPR-IB mRNA is strikingly restricted to the anterior olfactory nucleus
in the adult (Fig. 8). These results suggest that specific ligands for
BMPR-IB may play an important role in the adult in regulating the
function of the anterior olfactory nuclei. BMPR-IB mRNA is found in the
anterior olfactory nuclei as early as E19 with continued expression
throughout the postnatal period, suggesting that ligands for the
receptor regulate the cellular maturation of the nuclei as well.
Furthermore, at E12-E16, BMPR-IB mRNA is specifically expressed in the
anterior portion of the VZ, an area that may furnish progenitor cells
that populate the anterior olfactory nuclei. BMPR-IB transcripts are
also present at significant levels in the olfactory epithelium in
E12-E19 embryos, implying that specific BMPs may be involved in
the development of other aspects of the olfactory system as
well.
Expression of BMPR-I and BMPR-II mRNAs does not completely
overlap in many adult brain regions
BMPR-IA, BMPR-IB, and BMPR-II mRNAs do not fully overlap in the
adult brain. It has been reported that BMP type I and type II receptors
are both required for optimal signal transduction (Derynck, 1994;
Massague, 1996; ten Dijke et al., 1996). However, the differential
expression patterns of the two BMP type I receptors suggest that in
adult brain, BMPR-IA and BMPR-IB may convey strikingly different
physiological signals. If the current receptor model for signal
transduction of BMPs is correct, it is likely that additional subtypes
of BMPR-I and BMPR-II receptor subunits might exist. In this regard,
the conspicuous absence of BMPR-II in areas of BMPR-IB expression is
striking. This raises the possibility that another type II subunit may
exist in brain that is coexpressed with the type IB subunit. These
observations are very similar to previous findings of widespread
expression of type I TGF- receptors in developing mouse cortex
without detectable expression of TGF- type II receptors (Tomoda et
al., 1996); this too suggested to the authors that there may be other
as yet unidentified receptor components in brain. However unlike
TGF- receptors, homodimeric BMP receptors can also transduce
signals, albeit with lower efficiency than heterodimeric receptors
(Hogan, 1996). It is thus also possible that lower basal levels of
signaling may be sufficient in the adult brain to code for maintenance
functions and/or phasic plasticity events such as dendritic branching
(Lein et al., 1995). Finally, it is also possible that additional
receptors within the TGF- superfamily may form heterodimeric
signaling complexes with BMPR-IA, BMPR-IB, and BMPR-II.
| |
FOOTNOTES |
Received Feb. 2, 1998; accepted Feb. 18, 1998.
This work was supported by an Irma T. Hirschl Career Scientist Award
(M.F.M.) and grants from the Muscular Dystrophy Association (M.F.M.)
and the National Institutes of Health (NS35320 to M.F.M. and NS20013
and NS20778 to J.A.K.). We are grateful to Drs. Anthony Celeste and
John Wozney of Genetics Institute for providing molecular probes for
BMP receptor subunits and Dr. Kohei Miyazono of the Cancer Institute
(Tokyo, Japan) for providing affinity-purified antibodies to BMP type I
and type II receptor subunits. We thank Ms. Antoinette Barnecott for
her expert assistance in the preparation of this manuscript.
Correspondence should be addressed to John A. Kessler, Albert Einstein
College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461.
| |
REFERENCES |
-
Arkell R,
Beddington RSP
(1997)
BMP-7 influences pattern and growth of the developing hindbrain of mouse embryos.
Development
124:1-12[Abstract].
-
Birren SJ,
Anderson DJ
(1990)
A V-myc-immortalized sympathoadrenal progenitor cell line in which neuronal differentiation is initiated by FGF but not NGF.
Neuron
4:189-201[Web of Science][Medline].
-
Coughlin MD, Collins MB (1985) Nerve growth
factor-independent development of embryonic mouse sympathetic neurons
in dissociated cell culture Dev Biol 110:392-401.
-
D'Alessandro JS,
Vetz-Aldape J,
Wang EA
(1994)
Bone morphogenetic proteins induce differentiation in astrocyte lineage cells.
Growth Factors
11:53-69[Web of Science][Medline].
-
Dale L,
Howes G,
Price BM,
Smith JC
(1992)
Bone morphogenetic protein 4: a ventralizing factor in early Xenopus development.
Development
115:573-585[Abstract].
-
Derynck R
(1994)
TGF--receptor-mediated signaling.
Trends Biol Sci
19:548-553.
-
Dewulf N,
Verschueren K,
Lonnoy O,
Moren A,
Grimsby S,
Vande Spiegle K,
Miyazono K,
Huylebroeck D,
Ten Dijke P
(1995)
Distinct spatial and temporal expression patterns of two type I receptors for bone morphogenetic proteins during mouse embryogenesis.
Endocrinology
136:2652-2663[Abstract].
-
Fagan AM,
Zhang H,
Landis S,
Smeyne RJ,
Silos-Santiago I,
Barbacid M
(1996)
TrkA, but not trkC, receptors are essential for survival of sympathetic neurons in vivo.
J Neurosci
16:6208-6218[Abstract/Free Full Text].
-
Fann MJ,
Patterson PH
(1994)
Depolarization differentially regulates the effects of bone morphogenetic protein (BMP)-2, BMP-6 and activin A on sympathetic neuronal phenotype.
J Neurochem
63:2074-2079[Web of Science][Medline].
-
Francis PH,
Richardson MK,
Brickell PM,
Tickle C
(1994)
Bone morphogenetic proteins and a signaling pathway that controls patterning in the developing chick limb.
Development
120:209-218[Abstract].
-
Graham A,
Francis-West P,
Brickell P,
Lumsden A
(1994)
The signaling molecule BMP4 mediates apoptosis in the rhombencephalic neural crest.
Nature
372:684-686[Medline].
-
Graham A,
Koentges G,
Lumsden A
(1996)
Neural crest apoptosis and the establishment of craniofacial pattern: an honorable death.
Mol Cell Neurosci
8:76-83[Web of Science][Medline].
-
Gross RE,
Mehler MF,
Mabie PC,
Zang Z,
Santschi L,
Kessler JA
(1996)
Bone morphogenetic proteins promote astroglial lineage commitment by mammalian subventricular zone progenitor cells.
Neuron
17:595-606[Web of Science][Medline].
-
Hemmati-Brivanlou A,
Melton D
(1997)
Vertebrate embryonic cells will become nerve cells unless told otherwise.
Cell
88:13-17[Web of Science][Medline].
-
Hemmati-Brivanlou A,
Thomsen GH
(1995)
Ventral mesodermal patterning in Xenopus embryos: expression patterns and activities of BMP-2 and BMP-4.
Dev Genet
17:78-89[Web of Science][Medline].
-
Hogan BLM
(1996)
Bone morphogenetic proteins: multifunctional regulators of vertebrate development.
Genes Dev
10:1580-1584[Free Full Text].
-
Holley SA,
Neul JL,
Attisano L,
Wrana JL,
Sasai Y,
O'Connor MB,
DeRobertis EM,
Ferguson EL
(1996)
The Xenopus dorsalizing factor noggin ventralizes Drosophila embryos by preventing DPP from activating its receptor.
Cell
86:607-617[Web of Science][Medline].
-
Johnson RL,
Riddle RD,
Tabin CJ
(1994)
Mechanisms of limb patterning.
Curr Opin Genet Dev
4:535-542[Medline].
-
Kawakami Y,
Ishikawa T,
Shimabara M,
Tanda N,
Enomoto-Iwamoto M,
Iwamoto M,
Kuwana T,
Ueki A,
Noji S,
Nohno T
(1996)
BMP signaling during bone pattern determination in the developing limb.
Development
122:3557-3566[Abstract].
-
Kessler JA,
Adler J,
Bohn MC,
Black IB
(1981)
Substance P in principal sympathetic neurons: regulation by impulse activity.
Science
214:335-336[Abstract/Free Full Text].
-
Kingsley DM
(1994)
The TGF- superfamily: new members, new receptors, and new genetic tests of function in different organisms.
Genes Dev
8:133-146[Free Full Text].
-
Koenig BB,
Cook JS,
Wolsing DH,
Ting J,
Tiesman JP,
Correa PE,
Olson CA,
Pecquet AL,
Ventura F,
Grant RA,
Chen GX,
Wrana JL,
Massague J,
Rosenbaum JS
(1994)
Characterization and cloning for a receptor for BMP-2 and BMP-4 from NIH3T3 cell.
Mol Cell Biol
14:5961-5974[Abstract/Free Full Text].
-
Liem KF,
Tremml G,
Roelink H,
Jessell TM
(1995)
Dorsal differentiation of neural plate cells induced by BMP-mediated signals from epidermal ectoderm.
Cell
82:969-979[Web of Science][Medline].
-
Lein P,
Johnson M,
Guo X,
Rueger D,
Higgins D
(1995)
Osteogenic protein-1 induces dendritic growth in rat sympathetic neurons.
Neuron
15:597-605[Web of Science][Medline].
-
Lein P,
Guo X,
Hedges AM,
Rueger D,
Johnson M,
Higgins D
(1996)
The effects of extracellular matrix and osteogenic protein-1 on the morphological differentiation of rat sympathetic neurons.
Int J Dev Neurosci
14:203-215[Web of Science][Medline].
-
Liu F,
Ventura F,
Doody J,
Massagué J
(1995)
Human type II receptor for bone morphogenetic proteins (BMPs): extension of the two-kinase receptor model to the BMPs.
Mol Cell Biol
15:3479-3486[Abstract].
-
Massague J
(1996)
TGF beta signaling receptors, transducers, and Mad proteins.
Cell
85:947-950[Web of Science][Medline].
-
Mehler MF,
Kessler JA
(1995)
Cytokines and neuronal differentiation.
Crit Rev Neurobiol
9:419-446[Web of Science][Medline].
-
Mehler MF,
Mabie PC,
Zhang D,
Kessler JA
(1997)
Bone morphogenetic proteins in the nervous system.
Trends Neurosci
20:309-315[Web of Science][Medline].
-
Mishina Y,
Suzuki A,
Ueno N,
Behringer RR
(1995)
Bmpr encodes a type I bone morphogenetic protein receptor that is essential for gastrulation during mouse embryogenesis.
Genes Dev
9:3027-3037[Abstract/Free Full Text].
-
Mu X,
Silos-Santiago I,
Carroll SL,
Snider WD
(1993)
Neurotrophin receptor genes are expressed in distinct patterns in developing dorsal root ganglia.
J Neurosci
13:4029-4041[Abstract].
-
Nohno T,
Ishikawa T,
Saito T,
Hosokawa K,
Noji S,
Wosing DH,
Rosenbaum JS
(1995)
Identification of a human type II receptor for bone morphogenetic protein-4 that forms differential heteromeric complexes with BMP type I receptors.
J Biol Chem
270:22522-22526[Abstract/Free Full Text].
-
Piccolo S,
Sasai Y,
Lu B,
De Robertis EM
(1996)
Dorsoventral patterning in Xenopus: inhibition of ventral signals by direct binding of chordin to BMP-4.
Cell
86:589-598[Web of Science][Medline].
-
Reissmann E,
Ernsberger U,
Francis-West PH,
Rueger D,
Brickell PM,
Rohrer H
(1996)
Involvement of bone morphogenetic protein-4 and bone morphogenetic protein-7 in the differentiation of the adrenergic phenotype in developing sympathetic neurons.
Development
122:2079-2088[Abstract].
-
Rosenzweig BL,
Imamura T,
Okadome T,
Cox GN,
Yamashita H,
ten Dijke P,
Heldin CH,
Miyazono K
(1995)
Cloning and characterization of a human type II receptor for bone morphogenetic proteins.
Proc Natl Acad Sci USA
92:7632-7636[Abstract/Free Full Text].
-
Shah WM,
Groves AK,
Anderson DJ
(1996)
Alternative neural crest cell fates are instructively promoted by TGF beta superfamily members.
Cell
85:331-343[Web of Science][Medline].
-
Soderstrom S,
Bengtsson H,
Ebendal T
(1996)
Expression of serine/threonine kinase receptors including the bone morphogenetic factor type II receptor in the developing and adult rat brain.
Cell Tissue Res
286:269-279[Web of Science][Medline].
-
Song Q, Mehler MF, Kessler JA (1998) Bone morphogenetic
proteins induce apoptosis and growth factor dependence of cultured
sympathoadrenal progenitor cells. Dev Biol, in press.
-
Tanabe Y,
Jessell TM
(1996)
Diversity and pattern in the developing spinal cord.
Science
274:1115-1123[Abstract/Free Full Text].
-
ten Dijke P,
Yamashita H,
Sampath TK,
Reddi AH,
Estevez M,
Riddle DL,
Ichijo H,
Heldin CH,
Miyazono K
(1994a)
Identification of type I receptors for osteogenic protein-1 and bone morphogenetic protein-4.
J Biol Chem
269:16985-16988[Abstract/Free Full Text].
-
ten Dijke P,
Yamashita H,
Ichijo H,
Franzen P,
Laiho M,
Miyazono K,
Heldin CH
(1994b)
Characterization of type I receptors for transforming growth factor-beta and activin.
Science
264:101-104[Abstract/Free Full Text].
-
ten Dijke P,
Miyazomo K,
Heldin CH
(1996)
Signaling via hetero-oligomeric complexes of type I and type II serine/threonine kinase receptor.
Curr Opin Cell Biol
8:139-145[Web of Science][Medline].
-
Tomoda T,
Shirasawa T,
Yahagi YI,
Ishii K,
Takagi H,
Furiya Y,
Arai KI,
Mori H,
Muramatsu MA
(1996)
Transforming growth factor-beta is a survival factor for neonate cortical neurons: coincident expression of type I receptors in developing cerebral cortices.
Dev Biol
179:79-90[Web of Science][Medline].
-
Urist MR,
Mikulski A,
Lietze A
(1979)
Solubilized and insolubilized bone morphogenetic protein.
Proc Natl Acad Sci USA
76:1828-1832[Abstract/Free Full Text].
-
Varley JE,
Wehby RG,
Rueger DC,
Maxwell GD
(1995)
Number of adrenergic and islet-1 immunoreactive cells is increased in avian trunk neural crest cultures in the presence of human recombinant osteogenic protein-1.
Dev Dyn
203:434-447[Web of Science][Medline].
-
Varley JE,
Maxwell GD
(1996)
BMP-2 and BMP-4, but not BMP-6, increases the number of adrenergic cells which develop in quail trunk neural crest cultures.
Exp Neurol
140:84-94[Web of Science][Medline].
-
Wilson PA,
Hemmati-Brivanlou A
(1995)
Induction of epidermis and inhibition of neural fate by Bmp-4.
Nature
376:331-333[Medline].
-
Winnier G,
Blessing M,
Labosky PA,
Hogan BLM
(1995)
Bone morphogenetic protein-4 (BMP-4) is required for mesoderm formation and patterning in the mouse.
Genes Dev
9:2105-2116[Abstract/Free Full Text].
-
Wozney JM,
Rosen V,
Celeste AJ,
Mitsock LM,
Whitters MJ,
Kriz RW,
Hewick RM,
Wang EA
(1988)
Novel regulators of bone formation: molecular clones and activities.
Science
242:1528-1534[Abstract/Free Full Text].
-
Zhang H,
Bradley A
(1996)
Mice deficient for BMP2 are non viable and have defects in ammion/chorion and cardiac development.
Development
122:2977-2986[Abstract].
-
Zimmerman LB,
Dejesus-Escobar JM,
Harland RM
(1996)
The spemann organizer signal noggin binds and inactivates bone morphogenetic protein 4.
Cell
86:599-606[Web of Science][Medline].
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[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Olmsted-Davis, F. H. Gannon, M. Ozen, M. M. Ittmann, Z. Gugala, J. A. Hipp, K. M. Moran, C. M. Fouletier-Dilling, S. Schumara-Martin, R. W. Lindsey, et al.
Hypoxic Adipocytes Pattern Early Heterotopic Bone Formation
Am. J. Pathol.,
February 1, 2007;
170(2):
620 - 632.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Mazerbourg, K. Sangkuhl, C.-W. Luo, S. Sudo, C. Klein, and A. J. W. Hsueh
Identification of Receptors and Signaling Pathways for Orphan Bone Morphogenetic Protein/Growth Differentiation Factor Ligands Based on Genomic Analyses
J. Biol. Chem.,
September 16, 2005;
280(37):
32122 - 32132.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. E. Kendall, C. Battelli, S. Irwin, J. G. Mitchell, C. A. Glackin, and J. M. Verdi
NRAGE Mediates p38 Activation and Neural Progenitor Apoptosis via the Bone Morphogenetic Protein Signaling Cascade
Mol. Cell. Biol.,
September 1, 2005;
25(17):
7711 - 7724.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Chalazonitis, F. D'Autreaux, U. Guha, T. D. Pham, C. Faure, J. J. Chen, D. Roman, L. Kan, T. P. Rothman, J. A. Kessler, et al.
Bone Morphogenetic Protein-2 and -4 Limit the Number of Enteric Neurons But Promote Development of a TrkC-Expressing Neurotrophin-3-Dependent Subset
J. Neurosci.,
April 28, 2004;
24(17):
4266 - 4282.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Akiyoshi, K. Uchida, and S. Tateyama
Expression of Bone Morphogenetic Protein-6 and Bone Morphogenetic Protein Receptors in Myoepithelial Cells of Canine Mammary Gland Tumors
Vet. Pathol.,
March 1, 2004;
41(2):
154 - 163.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
U. Guha, W. A. Gomes, J. Samanta, M. Gupta, F. L. Rice, and J. A. Kessler
Target-derived BMP signaling limits sensory neuron number and the extent of peripheral innervation in vivo
Development,
March 1, 2004;
131(5):
1175 - 1186.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Viti, A. Gulacsi, and L. Lillien
Wnt Regulation of Progenitor Maturation in the Cortex Depends on Shh or Fibroblast Growth Factor 2
J. Neurosci.,
July 2, 2003;
23(13):
5919 - 5927.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Angley, M. Kumar, K. J. Dinsio, A. K. Hall, and R. E. Siegel
Signaling by Bone Morphogenetic Proteins and Smad1 Modulates the Postnatal Differentiation of Cerebellar Cells
J. Neurosci.,
January 1, 2003;
23(1):
260 - 268.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. J. Lein, H. N. Beck, V. Chandrasekaran, P. J. Gallagher, H.-L. Chen, Y. Lin, X. Guo, P. L. Kaplan, H. Tiedge, and D. Higgins
Glia Induce Dendritic Growth in Cultured Sympathetic Neurons by Modulating the Balance between Bone Morphogenetic Proteins (BMPs) and BMP Antagonists
J. Neurosci.,
December 1, 2002;
22(23):
10377 - 10387.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Wang, C.-F. Chang, M. Morales, J. Chou, H.-L. Chen, Y.-H. Chiang, S.-Z. Lin, J. L. Cadet, X. Deng, J.-Y. Wang, et al.
Bone Morphogenetic Protein-6 Reduces Ischemia-Induced Brain Damage in Rats
Stroke,
September 1, 2001;
32(9):
2170 - 2178.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. M. Panchision, J. M. Pickel, L. Studer, S.-H. Lee, P. A. Turner, T. G. Hazel, and R. D.G. McKay
Sequential actions of BMP receptors control neural precursor cell production and fate
Genes & Dev.,
August 15, 2001;
15(16):
2094 - 2110.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Nakashima, T. Takizawa, W. Ochiai, M. Yanagisawa, T. Hisatsune, M. Nakafuku, K. Miyazono, T. Kishimoto, R. Kageyama, and T. Taga
BMP2-mediated alteration in the developmental pathway of fetal mouse brain cells from neurogenesis to astrocytogenesis
PNAS,
April 25, 2001;
(2001)
101109698.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
N. Miao, B. Fung, R. Sanchez, J. Lydon, D. Barker, and K. Pang
Isolation and Expression of PASK, a Serine/Threonine Kinase, During Rat Embryonic Development, with Special Emphasis on the Pancreas
J. Histochem. Cytochem.,
October 1, 2000;
48(10):
1391 - 1400.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
S. H. Thang, M. Kobayashi, and I. Matsuoka
Regulation of Glial Cell Line-Derived Neurotrophic Factor Responsiveness in Developing Rat Sympathetic Neurons by Retinoic Acid and Bone Morphogenetic Protein-2
J. Neurosci.,
April 15, 2000;
20(8):
2917 - 2925.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J Shou, R. Murray, P. Rim, and A. Calof
Opposing effects of bone morphogenetic proteins on neuron production and survival in the olfactory receptor neuron lineage
Development,
January 12, 2000;
127(24):
5403 - 5413.
[Abstract]
[PDF]
|
|
|
|
|
|
|
L Lillien and H Raphael
BMP and FGF regulate the development of EGF-responsive neural progenitor cells
Development,
January 11, 2000;
127(22):
4993 - 5005.
[Abstract]
[PDF]
|
|
|
|
|
|
|
M. Srinivas, R. Rozental, T. Kojima, R. Dermietzel, M. Mehler, D. F. Condorelli, J. A. Kessler, and D. C. Spray
Functional Properties of Channels Formed by the Neuronal Gap Junction Protein Connexin36
J. Neurosci.,
November 15, 1999;
19(22):
9848 - 9855.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. F. McManus, L.-C. Chen, I. Vallejo, and M. Vallejo
Astroglial Differentiation of Cortical Precursor Cells Triggered by Activation of the cAMP-Dependent Signaling Pathway
J. Neurosci.,
October 15, 1999;
19(20):
9004 - 9015.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. C. Mabie, M. F. Mehler, and J. A. Kessler
Multiple Roles of Bone Morphogenetic Protein Signaling in the Regulation of Cortical Cell Number and Phenotype
J. Neurosci.,
August 15, 1999;
19(16):
7077 - 7088.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Nakashima, T. Takizawa, W. Ochiai, M. Yanagisawa, T. Hisatsune, M. Nakafuku, K. Miyazono, T. Kishimoto, R. Kageyama, and T. Taga
BMP2-mediated alteration in the developmental pathway of fetal mouse brain cells from neurogenesis to astrocytogenesis
PNAS,
May 8, 2001;
98(10):
5868 - 5873.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
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Top
Abstract
Introduction
