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The Journal of Neuroscience, November 15, 1998, 18(22):9335-9341
Persyn, a Member of the Synuclein Family, Has a Distinct Pattern
of Expression in the Developing Nervous System
Vladimir L.
Buchman1,
Hamish J. A.
Hunter1,
Luzia
G. P.
Pinõn1,
Jane
Thompson1,
Eugenia M.
Privalova1, 2,
Natalia N.
Ninkina1, 2, and
Alun M.
Davies1, 3
1 School of Biomedical Sciences, University of St.
Andrews, Bute Medical Buildings, St. Andrews, Fife KY16 9AJ, Scotland,
United Kingdom, 2 Institute of Gene Biology, Russian
Academy of Sciences, Moscow B-334, Russia, and 3 Neuropa
Limited, Robertson Building, Dumbarton Road, Glasgow G11 6NU, Scotland,
United Kingdom
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ABSTRACT |
The synucleins are a unique family of small intracellular proteins
that have recently attracted considerable attention because of their
involvement in human neurodegenerative diseases. We have cloned a new
member of the synuclein family called persyn. In contrast to other
synucleins, which are presynaptic proteins of CNS neurons, persyn is a
cytosolic protein that is expressed predominantly in the cell bodies
and axons of primary sensory neurons, sympathetic neurons, and
motoneurons. Northern blotting, in situ hybridization, Western blotting, and immunohistochemistry revealed that persyn mRNA
and protein are expressed in these neurons from the earliest stages of
axonal outgrowth and are maintained at a high level throughout life.
Persyn also becomes detectable in evolutionary recent regions of the
brain by adulthood.
Key words:
synucleins; neurodegenerative diseases; Alzheimer's
disease; Parkinson's disease; development of the nervous system; BCSG1; motoneurons; sensory neurons
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INTRODUCTION |
The accumulation of intracellular or
extracellular deposits in the form of plaques, tangles, or inclusion
bodies is a hallmark of many neurodegenerative diseases (Selkoe, 1994 ;
Kelly, 1996). These deposits are composed of one or more proteins and
peptides, and it has been suggested that certain minor polypeptide
components initiate aggregation of the main component (Wisniewski and
Frangione, 1992; Ma et al., 1994). One of such minor components found
in the plaques of Alzheimer's disease is NAC, an internal peptide of
NACP or  -synuclein, a member of a family of small proteins with an
unusual amino acid sequence and undefined functions (Maroteaux et al.,
1988; Maroteaux and Scheller, 1991; Uéda et al., 1993; Jakes et
al., 1994). The ability of -synuclein to self-aggregate and to bind
and induce the aggregation of amyloid A peptide (Iwai et al., 1995b;
Yoshimoto et al., 1995; Jensen et al., 1997; Paik et al., 1997, 1998)
is consistent with a potential role in promoting amyloid deposition in
Alzheimer's disease. The topography of synaptic function impairment in
Alzheimer's disease also correlates with the pattern of -synuclein
expression and its localization in synaptic terminals (Uéda et
al., 1993; Jakes et al., 1994; George et al., 1995; Iwai et al.,
1995a). Furthermore, synuclein (synelfin) has been implicated in at
least one form of learning and memory in zebrafinch (George et al.,
1995; Jin and Clayton, 1997). Both - and -synucleins are
expressed predominantly in evolutionary recent structures of the CNS
(Maroteaux and Scheller, 1991; Uéda et al., 1993, 1994; Jakes et
al., 1994).
Two mutations in the -synuclein gene have recently been reported to
be directly associated with an early-onset, autosomal-dominant form of
Parkinson's disease (Polymeropoulos et al., 1997; Kruger et al.,
1998). Although it has been shown that synucleins in solution are
random coiled or natively unfolded proteins, interactions with other
macromolecules in living cells could stabilize synucleins in particular
conformations (Weinreb et al., 1996; Kim, 1997; Paik et al., 1997,
1998; Davidson et al., 1998). Both mutations associated with
Parkinson's disease were predicted to change the secondary structure
of the -synuclein molecule in the way that increases the ability of
this protein to self-aggregate or form aggregates with other proteins
(Goedert, 1997; Heintz and Zoghbi, 1997; Nussbaum and Polymeropoulos,
1997; Polymeropoulos et al., 1997; Kruger et al., 1998). Consistently,
-synuclein, but not -synuclein, is found in the Lewy bodies of
cases of sporadic Parkinson's disease and dementia with Lewy bodies
(Spillantini et al., 1997). Accumulations of -synuclein are also
present in abnormal neurites that, like Lewy bodies, contain ubiquitin,
synaptophysin, and neurofilaments (Takeda et al., 1998). In contrast,
 -positive lesions (neurofibrillary tangles, Pick's bodies, neuropil
threads, and ballooned neurons) are synuclein-negative (Takeda et al., 1998). These data suggest that -synuclein is involved in specific aspects of the pathogenesis of neurodegeneration.
Here we report the characterization a member of the synuclein family
that we have called persyn. Detailed studies of the expression of
persyn mRNA and protein during normal embryonic and
postnatal development show that it has a very different pattern of
expression to that of other synucleins. This finding together with our
evidence implicating persyn in regulating the integrity of the
neurofilament network (Buchman et al., 1998) suggests that persyn could
be involved in modulating axonal architecture during development and in
the adult.
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MATERIALS AND METHODS |
Subtractive cloning. A subtractive cloning procedure
(Baka and Buchman, 1996) was used to isolate cDNAs corresponding to
mRNAs that are expressed at higher levels in embryonic day (E13) mouse trigeminal ganglia (TG) than in E13 mouse forebrain. Single-stranded cDNA was synthesized from E13 TG poly(A)+ RNA (leader) by
reverse transcriptase, and subtractive hybridization was performed with
photobiotinylated poly(A)+ RNA (driver). To subtract most
of the housekeeping sequences, E13 mouse liver and lung
poly(A)+ RNAs were used as the driver in two rounds of
hybridization. This was followed by two rounds of hybridization with
biotinylated poly(A)+ RNA from E13 mouse forebrain. cDNAs
that failed to form hybrids with biotinylated RNA were cloned as
described (Baka and Buchman, 1996). Among the clones isolated from the
resulting subtracted cDNA library were two independent overlapped
clones, representing persyn mRNA.
Miscellaneous cloning procedures. RNA extraction, isolation
of poly(A)+ RNA, preparation of hybridization probes,
Northern, in situ, and Southern hybridizations, and library
screening were performed as described (Buchman et al., 1992, 1994).
Full-length cDNAs were isolated from an E13 mouse trigeminal cDNA
library constructed in the  ZAPII vector and a cDNA library from
juvenile human brainstem (Stratagene, La Jolla, CA).
Anti-persyn antibody. Rabbits were immunized with the 15-mer
C-terminal peptide of mouse persyn conjugated with keyhole limpet hemocyanin (Calbiochem, San Diego, CA) activated by MBS (Sigma, St. Louis, MO) (Harlow and Lane, 1988). Monospecific antibodies were
purified from the antisera by affinity chromatography using the antigen
bound to NHS-activated columns (Supelco, Bellefonte, PA). The
anti-mouse monospecific antibody was used at dilutions of 1:500 for
Western blot/ECL detection of persyn in total cell protein samples. In
some experiments 10 ml of diluted antibody was preincubated with 15 µg of the recombinant mouse persyn protein at room temperature for 2 hr.
Protein extraction and Western blotting/ECL detection.
Dissected trigeminal ganglia were homogenized directly in SDS-PAGE loading buffer (Laemmli, 1970) and incubated for 5 min in a boiling water bath. Total protein concentrations were measured by the dotMETRIC
assay (Geno Technology, St. Louis, MO). Fifteen micrograms of total
protein were used for SDS-PAGE (Laemmli, 1970). Electroblotting on
Hybond-PVDF membranes was performed in Tris-glycine-methanol buffer
as recommended by the membrane supplier (Amersham). Rainbow markers
from Amersham were used as protein size standards. After washing with
PBS, the membrane was blocked for 1 hr at room temperature in 4%
skimmed milk-0.05% Tween 20-PBS. The same buffer was used for
incubations with primary and secondary antibodies and washes. The final
two washes were in 0.1% Tween 20-PBS. ECL detection was performed as
recommended by the Amersham protocol.
Subcellular fractionation. Fractionation of homogenates of
the spinal cords, trigeminal ganglia, midbrains, and hindbrains of
adult mice was performed as described previously (Cotman and Taylor,
1972; Jones and Matus, 1974). Aliquots of the homogenate (hmg), 1000 gm
supernatant (cyt), 14,000 gm supernatant (pmt), 14,000 gm pellet (msk),
120,000 gm supernatant (pmc), 120,000 gm pellet (mcs), mitochondrial
(mt), and synaptosomal (syn) fractions of the sucrose gradient were
mixed with equal volumes of 2× loading buffer (Laemmli, 1970) and
incubated for 5 min in a boiling water bath. Twenty micrograms of total
protein from each fraction were used for SDS-PAGE followed by Western
blotting and detection as described above.
Immunohistochemistry. Fifteeen micrometer cryosections were
fixed in acetone at  20°C for 10 min and were dried and blocked with
5% goat serum in TBT (20 mM Tris HCl, pH 7.5; 150 mM NaCl; and 0.1% Triton X-100) followed by incubation
with the anti-persyn antibody (1:50) at 4°C for 16 hr in 1% goat
serum-TBT. Sections were washed with 1% goat serum-TBT and processed
as recommended by suppliers of secondary HRP-conjugated antibody
(Sigma). The same procedure was used for cultured neurons fixed on the
Petri dish with cold acetone-methanol mixture (1:1). FITC-conjugated secondary anti-rabbit antibody (Jackson ImmunoResearch, West Grove, PA)
was used for detection.
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RESULTS |
Cloning of persyn cDNA
persyn cDNA clones were isolated using the subtractive
cloning procedure described in Materials and Methods. Using the insert of the longest clone as a probe for Northern hybridization, a single
0.8 kb transcript was detected only in neural tissues of the E13 mouse
embryo. The level of this transcript was much higher in sensory ganglia
(trigeminal and dorsal root ganglia) than in CNS, reflecting the way in
which the clone was isolated (Fig. 1a). Similar results were
obtained when the mouse persyn probe was used for
hybridization with RNAs from newborn rat tissues (Fig.
1b).
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Figure 1.
Persyn transcripts in mouse and rat
tissues. Northern hybridization of 5-20 µg of total RNA extracted
from various tissues of E13 mouse embryos (a) and
newborn rat (b) with a nick-translated
persyn probe. After stripping off the probe, the same
filter was hybridized with a nick-translated cDNA fragment encoding the
mouse L27 ribosomal protein to provide an indication of the amount of
total RNA from each tissue present on the filter. TG,
Trigeminal ganglia; DRG, dorsal root ganglia.
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To isolate a full-length persyn cDNA, an E13 mouse
trigeminal ganglion cDNA library was constructed and screened with the mouse persyn cDNA probe. Four overlapping cDNA clones were
isolated from this screen, and sequence analysis revealed that the
longest of these included 5' untranslated, coding and 3' untranslated (with a characteristic polyadenylation signal) regions (GenBank accession number AF017255). Two human persyn cDNA clones
were isolated by screening a juvenile brainstem cDNA library with the mouse probe at low stringency. One of these clones included the complete coding region for human persyn protein (GenBank accession number AF017256).
Persyn is a member of the synuclein family
Figure 2 shows the high degree of
homology between persyn and other synucleins. All of the EKTKEGV
repeats, which are characteristic of the family (Maroteaux et al.,
1988; Maroteaux and Scheller, 1991; Uéda et al., 1993; Jakes et
al., 1994), are conserved in persyn. The C-terminal region of persyn is
highly negatively charged (12 of last 26 amino acids are glutamic or
aspartic acid residues), and this region has no obvious homology with
other synucleins. The C-terminal amino acid sequences in most
synucleins are variable and negatively charged (with the exception of
rat synuclein 2, which is charged positively). The sequence of persyn
is closely related to the sequence of the founder of the family,
Torpedo synuclein (Maroteaux et al., 1988). Two previously published
EST sequences that are most closely related to persyn [rat
synuclein-like (Akopian and Wood, 1995) and human BCSG1 (Ji et al.,
1997)] have multiple substitutions of amino acid that are highly
conserved in the synuclein family (Fig. 2). These substitutions are
probably resulted from Taq polymerase and sequence
errors.
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Figure 2.
Homologies between mouse persyn and other members
of the synuclein family. Alignment of persyn and other members of the
synuclein family. Amino acids identical in all or in all but one
members of the family are shown in white on black
background. Gaps (-) were introduced for better alignment.
Sequence accession numbers: Torpedo californica synuclein, P37379; rat
synucleins 1, 2, 3, products of alternative splicing of the same gene
S73007, S73008, S73009; human -synuclein (synuclein 1/NACP), L36674;
human -synuclein (synuclein 2), S69965; bovine synuclein, P33567;
zebrafinch synelfin, L33860; rat synuclein-like, X86789; human BCSG1,
AF010126; mouse persyn, AF017255; human persyn, AF017256.
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persyn mRNA is expressed in embryonic sensory
and motoneurons
A digoxygenin-labeled cRNA probe was used for whole-mount in
situ hybridization of E11 mouse embryos. Consistent with the results of Northern hybridization, signals were detected in all dorsal
root ganglia (DRG) and cranial sensory ganglia and in two strips within
the spinal cord (Fig. 3). For precise
localization of persyn-positive cells, in situ
hybridization on cryosections was performed. At E10, before the
formation of DRG, signals were only detected in the ventral horns of
the spinal cord in transverse sections (Fig.
4a). In E11 and E12 embryos,
when DRG have become evident, signals were additionally detected in DRG
(Fig. 4a). Signals were also observed in all cranial sensory
ganglia and in the regions of the hindbrain and midbrain where
branchiomotor and somatomotor neurons are located. This is illustrated
in the coronal section of an E11 head where hybridization signals in the motoneurons of oculomotor nucleus can be seen (Figs.
4b,c). On transverse sections of postnatal day 9 (P9) mice, persyn mRNA was detected in ventral horn
motoneurons, most if not all DRG neurons and in the neurons of
paravertebral sympathetic chain (Fig.
5a,b).
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Figure 3.
Top Left. Whole-mount in situ
hybridization of mouse embryos. E11 mouse embryos were hybridized with
a DIG-labeled antisense cRNA persyn probe.
TG, Trigeminal ganglion; VG, vestibular
ganglion; GG, geniculate ganglion; JG,
jugular ganglion; NG, nodose ganglion;
DRG, dorsal root ganglia; MN, motoneurons
in ventral horns of the spinal cord.
Figure 4.
Bottom left. Detection of
persyn expression in mouse embryonic tissues by
in situ hybridization. a, Transverse
sections at the level of the forelimb buds of E10,
E11, and E12 embryos were hybridized with
a DIG-labeled antisense cRNA persyn probe.
vsc, Motoneurons in ventral horns of the spinal cord;
drg, dorsal root ganglia. Scale bar, 1 mm.
b, Coronal section of E11 mouse embryo head.
tg, Trigeminal ganglion; vg, vestibular
ganglion; gg, geniculate ganglion; jg,
jugular ganglion; pg, petrosal ganglion;
ov, otic vesicle; hb, hindbrain;
fb, forebrain; omn, oculomotor nucleus.
Scale bar, 0.2 mm. c, A higher magnification of a
coronal section of an E11 mouse embryo head showing strong labeling in
the oculomotor nucleus and trigeminal ganglion. Scale bar, 80 µm.
Figure 5.
Top right. Detection of
persyn expression in mouse postnatal neurons.
a, In situ hybridization of transverse
section through the thorax of a P9 mouse neonate with a DIG-labeled
antisense cRNA persyn probe. VSC,
Motoneurons of the ventral horns of the spinal cord;
DRG, dorsal root ganglion; PSG,
paravertebral sympathetic ganglia. Scale bar, 0.2 mm. b,
Higher magnification of part of the DRG illustrated in the panel
(a) showing the cytoplasmic labeling of neurons.
c, Immunocytochemical localization of persyn protein in
processes and growth cones of cultured mouse DRG neurons. Scale bar, 20 µm.
Figure 6.
Bottom right. Detection of
persyn expression in adult brain by in
situ hybridization. In situ hybridization of
parasagittal sections of adult rat brain with a DIG-labeled antisense
cRNA persyn probe. Labeled neurons within the trigeminal motor nucleus
(a), frontal cortex (b),
CA1, and CA3 regions of hippocampus and cerebellar cortex are shown.
Scale bars: a, b, 0.1 mm;
c, d, 0.2 mm. The whole brain images
demonstrating absence of hybridization with persyn cDNA sense probe
(e) and specific hybridization with persyn cDNA
antisense probe (f) are shown.
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Persyn in adult brain
Although no persyn protein and mRNA were detected in embryonic and
neonatal forebrain structures (Figs. 1, 3, 4) accumulation of persyn
protein in the cerebral cortex with age was observed using Western
blotting (Buchman et al., 1998). To reveal which kinds of cells express
persyn mRNA in adult brain we localized persyn
mRNA by in situ hybridization in parasagital cryosections of
rat brain. The strongest hybridization signals were detected in the
motoneurons of the brainstem (Fig.
6a). However, substantial levels of expression were found in neurons in many other brain regions
including: Purkinje and granule cell layers of the cerebellar cortex
and the deep cerebellar nuclei, thalamus, hypothalamus, olfactory
bulbs, CA1, CA2, CA3, and CA4 regions of the hippocampus, and all
neuron-containing layers of the cerebral cortex in all regions examined
(Fig. 6).
Regulation of persyn expression during development of mouse
trigeminal ganglion
Because the normal development of the embryonic mouse trigeminal
ganglion is known in detail (Davies and Lumsden, 1984, 1986), the
expression of persyn was studied in this ganglion at closely staged intervals throughout development to ascertain when persyn is first expressed and how its expression changes during development. Quantitative Northern hybridization revealed that persyn
mRNA is expressed in the trigeminal ganglion from the earliest stages of its formation. There was a marked increase in expression between E10
and E12, the stage during which the earliest axons are growing to their
targets. This high level of expression was maintained into adulthood
(Fig. 7A). Western blotting
revealed that the developmental time course of persyn protein
expression in the trigeminal ganglion (Fig. 7B) was
consistent with the time course of persyn mRNA expression observed in Northern blotting studies.
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Figure 7.
Developmental time course of persyn expression in
the trigeminal ganglia. a, Graph showing the changes in
the level of persyn mRNA in the trigeminal ganglia
during development. The mean ± SEM of the persyn mRNA levels
relative to GAPDH mRNA levels are shown (n = 3).
b, Western blot showing the developmental changes in the
level of persyn protein the trigeminal ganglia. Equal amounts (15 µg)
of total protein from embryonic (E) or newborn
(P0) mouse trigeminal ganglia were run in each
lane.
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Persyn is cytosolic protein localized in neuronal cell bodies
and processes
Specific polyclonal antibodies (SK23) were raised in rabbits
against a synthetic C-terminal 15-mer peptide (PS) of mouse persyn. These antibodies detected a single band of ~16 kDa on Western blots
that disappeared if the antibodies were preincubated with excess of
either the C-terminal peptide or a recombinant persyn protein (Fig.
8a,b; data
not shown). The mobility of the protein was 3 kDa more than expected
from the persyn sequence, but similar decreases in mobility have been
reported for other synucleins (Jakes et al., 1994; Weinreb et al.,
1996). This antibody does not cross-react with either recombinant -
or -synucleins and does not detect native - and -synucleins in
the newborn mouse cerebral cortex (Fig. 8c), the
tissue where these proteins could be easy detected with specific
antibody (Shibayama-Imazu et al., 1993; Hsu et al., 1998).
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Figure 8.
Specificity of anti-mouse persyn
antibody. Western blot/ECL detection of persyn protein with
affinity-purified SK-23 antibody. a, b,
Equal amounts (15 µg) of total protein from P0 mouse heart and
hindbrain and E13 and E17 trigeminal ganglia were run in each lane.
Western blots were probed with SK-23 antibody (a)
or SK-23 antibody preincubated with recombinant mouse persyn
protein (b) as described in Materials and
Methods. c, Fifteen micrograms of total protein from
newborn (P0) mouse trigeminal ganglia (the tissue with a high
level of persyn expression) and cerebral cortex (the tissue where
persyn expression could not be detected but with substantial levels of
- and -synucleins expression) and 0.5 µg of recombinant -
and -synucleins were analyzed.
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Western blotting was used to study the distribution of persyn in
subcellular fractions of the mouse spinal cord, trigeminal ganglion,
midbrain, and hindbrain. Persyn protein was detected only in the
cytosolic fractions but not in the particulate and cytoskeletal
fractions (Fig. 9). Immunohistochemistry
using the SK23 antibodies has shown that persyn is localized in the
cell bodies and axons of sensory neurons (Buchman et al., 1998).
Likewise, immunocytochemical detection of persyn in cultured DRG
neurons showed that persyn is present in the cell bodies, processes,
and growth cones of these neurons (Fig. 5c) and that it is
expressed in both large NT3-dependent and small NGF-dependent neurons
(Fig. 5b; data not shown).
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Figure 9.
Persyn in subcellular fractions from mouse spinal
cord. Western blot showing the distribution of persyn protein in
subcellular fractions of the adult mouse spinal cord.
hmg, Homogenate; cyt, 1000 gm
supernatant; pmt, 14,000 gm supernatant;
pmc, 120,000 gm supernatant; mcs, 120,000 gm pellet; msk, 14,000 gm pellet; mt,
mitochondrial fraction; syn, synaptosomal fraction.
Equal amounts (15 µg) of total protein from each fraction were run in
each lane. Very similar results were obtained with subcellular
fractions of trigeminal ganglia, midbrain, and hindbrain.
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DISCUSSION |
We have cloned and characterized a member of the synuclein family
that has a distinctive pattern of expression in the developing and
mature nervous system. Persyn shares the main structural features of
the family, namely, conservation of all EKTKEGV repeats and the
C-terminal part of the molecule is negatively charged. In contrast to
- and -synucleins, which are predominantly expressed in the
cerebral cortex and other forebrain structures (Uéda et al.,
1993, 1994; Jakes et al., 1994; George et al., 1995; Iwai et al.,
1995a), persyn is abundant in primary sensory neurons and motoneurons.
These are the only neurons that express persyn in embryos, although
persyn is expressed in sympathetic ganglia in neonates and many
different kinds of neurons throughout the brain in the adult.
Immunocytochemical studies suggest that persyn is distributed diffusely
throughout the cell body and axons of sensory neurons and motoneurons.
This agrees with the results of subcellular fractionation of mouse
neuronal tissues that show that persyn is a predominantly cytosolic
protein that is not or very loosely associated with cytoskeletal or
vesicular fractions. This is similar to the intracellular distribution
of human and rat synucleins and zebrafinch synelfin (George et al.,
1995; Irizarry et al., 1996). However, both - and -synuclein are
apparently predominantly presynaptic proteins in the cerebral cortex
(Iwai et al., 1995a; Masliah et al., 1996; Irizarry et al., 1996).
Although the physiological role of synucleins is unknown, it has been
suggested that specific mutations could change the structural and
functional properties of these proteins, triggering mechanisms that
lead to neurodegeneration (Goedert, 1997; Heintz and Zoghbi, 1997;
Nussbaum and Polymeropoulos, 1997; Polymeropoulos et al., 1997).
Interestingly, human persyn has a threonin in position 53, which makes
it structurally similar to mutated (Ala53Thr) -synuclein found in
some families with hereditary early-onset form of Parkinson's disease
(Polymeropoulos et al., 1997). However, what kind of structural and
functional consequences this difference could have is not clear.
Although it was speculated that Ala53Thr substitution disrupts an
-helix and extends a -sheet in the predicted structure in this
part of the molecule (Polymeropoulos et al., 1997), all experimental
attempts to resolve the secondary structure of the wild-type
-synuclein have led to conclusion that synucleins are "natively
unfolded" or random coiled in solution (Weinreb et al., 1996; Kim,
1997). However, it was shown that binding to synthetic membranes
stabilizes -synuclein in an -helical conformation (Davidson et
al., 1998). Perhaps synucleins exist in different conformations in
cells depending on their association with membranes, and it is possible
that soluble and membrane-bound forms could have different functions.
We have recently shown that persyn plays role in regulating the
integrity of the neurofilament network in cultured sensory neurons
(Buchman et al., 1998). In this respect, it is interesting that the
onset of persyn expression in trigeminal sensory neurons coincides with
the stage when axons are starting to grow to their targets and remains
high throughout the life, raising the possibility that persyn plays a
role in regulating axonal growth and influencing axonal morphology.
The effect of persyn on neurofilaments could be important in
neurodegenerative conditions especially given the accumulation of
persyn in neurons of the cerebral cortex with age and its localization within axons. Loss of nerve cell processes is believed to contribute significantly to cerebral atrophy in neurodegenerative diseases. Understanding the pathophysiology of this process is important for
developing therapeutic strategies to prevent loss of neuronal connectivity. In further work it will be important to investigate the
possible contribution of persyn to the loss of neurons and their
processes in neurodegenerative diseases.
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FOOTNOTES |
Received June 19, 1998; revised Aug. 18, 1998; accepted Aug. 28, 1998.
This work was supported by a grant from the Wellcome Trust.
Correspondence should be addressed to Dr. V. L. Buchman, School of
Biological and Medical Sciences, University of St. Andrews, Bute
Medical Buildings, St. Andrews, Fife, KY16 9AJ, United Kingdom.
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Top
Abstract
Introduction
