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The Journal of Neuroscience, July 23, 2003, 23(16):6576-6585
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Abnormal Dysbindin Expression in Cerebellar Mossy Fiber Synapses in the mdx Mouse Model of Duchenne Muscular Dystrophy
Roy V. Sillitoe,1 Matthew A. Benson,2 Derek J. Blake,2 andRichard Hawkes1 1Department of Cell Biology and Anatomy, and Genes and Development Research Group, Faculty of Medicine, The University of Calgary, Calgary, Alberta T2N 4N1, Canada, and 2Department of Pharmacology, University of Oxford, Oxford, OX1 3QT, United Kingdom
Abstract
Top
Abstract
Introduction
The dystrophin-associated protein complex (DPC), comprising sarcoglycans, dystroglycans, dystrobrevins, and syntrophins, is a component of synapses both in muscle and brain. Dysbindin is a novel component of the DPC, which binds to-dystrobrevin and may serve as an adaptor protein that links the DPC to an intracellular signaling cascade. Disruption of the DPC results in muscular dystrophy, and mutations in the human ortholog of dysbindin have been implicated in the pathogenesis of schizophrenia. In both cases, patients also present with neurological symptoms reminiscent of cerebellar problems. In the mouse cerebellum, dysbindin immunoreactivity is expressed at high levels in a subset of mossy fiber synaptic glomeruli in the granular layer. Lower levels of dysbindin immunoreactivity are also detected in Purkinje cell dendrites. In the cerebellar vermis, dysbindin-immunoreactive glomeruli are restricted to an array of parasagittal stripes that bears a consistent relationship to Purkinje cell parasagittal band boundaries as defined by the expression of the respiratory isoenzyme zebrin II/aldolase c. In a mouse model of Duchenne muscular dystrophy, the mdx mutant, in which dystrophin is not expressed, there is a dramatic increase in the number of dysbindin-immunoreactive glomeruli in the posterior cerebellar vermis. Moreover, the topography of the terminal fields is disrupted, replacing the stripes by a homogeneous distribution. Abnormal synaptic organization in the cerebellum may contribute to the neurological problems associated with muscular dystrophy and schizophrenia.
Key words: Purkinje cell; dystrophin; zebrin II; unipolar brush cell; choline acetyltransferase; schizophrenia
Introduction
The dystrophin-associated protein complex (DPC) of the neuromuscular junction is comprised of three distinct components: the sarcoglycan, dystroglycan, and cytoplasmic complexes, which include- and
). Biochemical analysis of postsynaptic density-enriched synaptosomal proteins and immunocytochemical localization of several DPC members [e.g., dystrophin (Lidov et al., 1990
Recently, a novel member of the DPC, the 40 kDa coiled-coil protein dysbindin, was identified by using a yeast two-hybrid screen for interacting proteins with
The neurological symptoms of patients with both muscular dystrophy and schizophrenia include ocular control abnormalities characteristic of cerebellar problems [muscular dystrophy (Liu et al., 2001; Anderson et al., 2002
Materials and Methods
Animals: perfusion and sectioning. Animal procedures conformed to institutional regulations and the Guide to the Care and Use of Experimental Animals from the Canadian Council of Animal Care. Normal adult CD1, Balb/c, and C57BL/6 mice (20-30 gm; +/+, no strain-specific differences noted) were obtained from Charles River Laboratories (St. Constant, Quebec, Canada), and C57BL/10ScSn-Dmdmdx/J hemizygous male (mdx/Y) and homozygous female mutant mice (mdx/mdx) were obtained from The Jackson Laboratory (Bar Harbor, ME) and maintained in the Animal Resource Centre at the University of Calgary. Mice were deeply anesthetized with an intraperitoneal injection of 100 mg/kg somnotol. The mice were then killed by transcardiac perfusion with an initial rinse using ice-cold 0.9% saline followed by either ice-cold Bouin's fixative or 4% paraformaldehyde in PBS, pH 7.2 (Sigma, St. Louis, MO). The brains were removed and immersion-fixed for an additional 24 hr at 4°C. The cerebella were then cryoprotected through a series of buffered 10% (2 hr), 20% (2 hr), and 30% (overnight) sucrose solutions, serially sectioned in the transverse plane at 40 µm, and mounted on glass slides.3-Acetylpyridine injections. Adult Balb/c mice (n = 6: 20-25 gm) were given an intraperitoneal injection of 3-acetylpyridine (50 mg/kg, i.p., diluted 1:100 in PBS; Sigma) and monitored closely for 3-5 hr after treatment for respiratory difficulties (control animals, n = 3, were injected intraperitoneally with an equal volume of PBS). Animals were killed when ataxia became evident (7-11 d) and the cerebella were fixed and sectioned as described above. Two parallel series of sections were collected: one was stained for cresyl violet and the other processed for fluorescence anti-dysbindin/anti-calbindin double immunohistochemistry (see below).
Immunohistochemistry. All antibodies were diluted in 10% normal goat serum in PBS. Mouse monoclonal anti-zebrin II was used directly from spent hybridoma culture medium diluted 1:1000 (Brochu et al., 1990
Sections were rinsed three times in PBS for 5 min each, incubated in 30% H2O2 for 10 min at room temperature, and again rinsed three times in PBS for 5 min. Sections were then blocked in 10% normal goat serum in PBS for 2 hr at room temperature. After blocking, sections were incubated overnight at room temperature in primary antibodies. After three 5 min rinses in PBS, sections were incubated in either HRP-conjugated goat anti-rabbit (diluted 1:200) or Vectastain Elite biotinylated rabbit IgG (1:300; Vector Laboratories, Burlingame, CA) as appropriate. Staining was visualized by incubation in 0.5 mg/ml diaminobenzidine (Sigma), 0.5 µl/ml 30% H2O2 in PBS until the desired color intensity was achieved. Sections were then dehydrated and mounted in Entellan (EM Science, Gibbstown, NJ). For double labeling the tissue was incubated in both primary antibodies overnight at room temperature, rinsed, and then incubated for 2 hr in a mixture of Alexa-546-conjugated goat anti-rabbit Ig (Molecular Probes, Eugene, OR) and cyanine 2 (Cy2)-conjugated donkey anti-mouse Ig (Jackson ImmunoResearch Laboratories, West Grove, PA), both diluted 1:1000. After several rinses in PBS, sections were coverslipped in nonfluorescing mounting medium (Fluorsave Reagent, Calbiochem, La Jolla, CA).
Photomicrographs were captured either with a PhotoMetrics (Huntington Beach, CA) Quantix digital camera running under V for Windows or a SPOT Cooled Color digital camera (Diagnostic Instruments, Sterling Heights, MI). For confocal microscopy, an Olympus (Tokyo, Japan) Fluoview BX50 microscope was used and Z-stacked images (10 layers, each 2 µm in depth) were captured by using Fluoview software. Montages were assembled in Adobe Photoshop (Adobe Systems, San Jose, CA). The images were cropped and corrected for brightness and contrast but not otherwise manipulated.
Western blotting. Western blot analysis of both wild-type and mdx tissue was performed using a conventional protocol. Cerebellar homogenates were separated using PAGE according to Towbin et al. (1979
Results
Dysbindin expression in the mouse cerebellum
Western blot analysis reveals that dysbindin is expressed at high levels in the mouse cerebellum (Fig. 1A). Consistent with previous data from whole brain (Benson et al., 2001
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Figure 1. A, Western blot analysis of dysbindin in the cerebella of adult mice. Anti-dysbindin staining of an extract from adult mouse tibialis anterior (M) reveals a single immunoreactive band at an apparent molecular weight of 50 kDa. An anti-dysbindin immunoreactive protein running at the same apparent molecular weight is detected in the cerebellum (Cb) at levels ninefold higher than those found in muscle. In addition, a second, weaker, lower apparent molecular weight band is seen irregularly in the cerebellar extract, that is never detected in muscle. B, Immunoperoxidase staining of a transverse section through the hemispheres with anti-dysbindin shows dysbindin immunoreactivity in the somata (arrowhead) and dendrites (arrow) of Purkinje cells. gl, Granular layer; ml, molecular layer, pcl, Purkinje cell layer. C, Transverse section through the posterior lobe vermis (lobules VIII-X) immunoperoxidase stained with anti-dysbindin. Reaction product is deposited uniformly in the molecular layer (ml) and as clusters of immunoreactive punctae in the granular layer. There is no staining of the white-matter tracts (wm). D, Immunoperoxidase staining of a transverse section through lobule IX of the cerebellar vermis with anti-dysbindin shows heterogeneous dysbindin immunoreactivity in the granular layer, with a cluster of immunoreactive mossy fiber terminals flanked by regions of granular layer with little or no reaction product. Staining is weak in the Purkinje cell layer and molecular layer. E, Dysbindin immunoreactivity in the granular layer is associated with large mossy fiber synaptic glomeruli. Occasionally, axon-like processes are observed (arrowhead), terminating in prominent synaptic rosettes. F, Double immunofluorescence staining of lobule IX for synaptophysin (green) and dysbindin (red) shows that dysbindin co-localizes with a subset of synaptophysin immunoreactive terminals (orange) in the granular layer (synaptophysin-positive/dysbindin-negative synapses are green). Scale bars: A, B, D, 100 µm; C, 500 µm; E, 10 µm; F, 50 µm.
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Figure 7. Dysbindin expression is abnormal in mossy fiber synaptic glomeruli of the mdx mouse cerebellum. A, B, Immunoperoxidase-stained transverse sections through lobule IX of the control (A, +/+) and dystrophic (B, mdx) mice. The dashed line indicates the midline. The 1 and 2 clusters of dysbindin-immunoreactive glomeruli are labeled in A. The same clusters are difficult to discern in the mdx mouse (B) because the intervening granular layer now has numerous immunoreactive profiles (B). A, inset, Western blot analysis comparing dysbindin expression in protein extracts from the cerebella of mdx mice versus wild-type littermates. In both the +/+ and mdx, a prominent band was detected at 50 kDa: densitometric scanning revealed no significant difference between the two (the lower-molecular-weight antigen seen in Fig. 1 was not detected in this sample). C, D, The regions of granular layer outlined by the rectangles in A and B, showing ectopic dysbindin immunoreactivity in the mdx mouse. E, Histograms illustrating the number of glomeruli in six individuals: three mdx and three +/+ (control littermates). The bins represent the mean ± SE numbers of dysbindin immunoreactive profiles from three 40 µm transverse sections from each individual, taken from ventral lobule IX immunoperoxidase stained with anti-dysbindin. The scale is in millimeters. There is a more than threefold increase in the number of immunoreactive glomeruli in mdx. Furthermore, in +/+, at least five parasagittal clusters are evident (1, at the midline; 2 and 3, laterally to either side), whereas in mdx, distinct clusters are obscured. Scale bars: A, B (in B), 250 µm; C, D (in D), 50 µm.
Peroxidase immunohistochemistry with anti-dysbindin heavily deposits peroxidase reaction product in punctate patches in the granular layer of the adult mouse cerebellum (Fig. 1B,C,D). In addition, dysbindin-like immunoreactivity is seen at lower levels in Purkinje cell somata (Fig. 1B, arrowhead) and dendrites (Fig. 1B, arrow) in the molecular layer. The intensity of Purkinje cell immunoreactivity varies significantly between lobules (Fig. 1, compare B and D). It is unclear whether differential immunoreactivity in the granular and Purkinje cell layers is attributable to differences in the level of expression or if it reflects the accessibility of the dysbindin epitope. Seen at high magnification, the granular layer staining is associated with mossy fiber synaptic glomeruli, a complex synaptic structure comprising a large mossy fiber presynaptic terminal, several postsynaptic granule cell dendrites, and inhibitory inputs from Golgi interneurons (Fig. 1E). To confirm that dysbindin immunoreactivity in the granular layer is associated with synapses, sections were double-immunofluorescence-stained for dysbindin and synaptophysin (Fig. 1F). Most mammalian CNS synapses are enriched in the 38 kDa synaptic vesicle protein synaptophysin (Wiedenmann and Franke, 1985
Granular layer heterogeneity
Because dysbindin expression is restricted to a subset of mossy fiber glomeruli, we undertook a systematic analysis of their topography. Patterns of gene expression, afferent connectivity and cerebellar mutations affecting neonate and adult mice reveal a set of boundaries that divides the cerebellum into four transverse zones: the anterior zone (AZ;
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Figure 2. Transverse sections through the vermis immunoperoxidase stained for dysbindin show numerous mossy fiber synaptic glomeruli in the granular layer of lobule II (A), few present in lobule VI (B), and heavy labeling in lobule X (C). In the AZ and the NZ, dysbindin-immunoreactive glomeruli are distributed heterogeneously to form clusters (numbered 1-3: numbering is specific to individual transverse zones, and there is no reason to assume that stripes are contiguous across transverse zone boundaries). In C, the white brackets outline cluster 1 in ventral lobule IX and dorsal lobule X. pcl, Purkinje cell layer. Scale bar, 250 µm.
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Figure 3. A, Dysbindin immunoreactive profiles in the granular layer of the AZ (lobules II-V) terminate in parasagittal stripes. Immunolabeled mossy fiber glomeruli in the granular layer of the AZ (lobules II-V) were plotted from every second 40 µm section through the vermis (for additional details, see Ozol et al., 1999
To relate the topography of the dysbindin-immunoreactive mossy fiber terminal fields to the better-understood Purkinje cell compartmentation, we have compared the stripes of dysbindin immunoreactive mossy fiber synaptic glomeruli to the parasagittal stripes of Purkinje cells, as revealed by two adult stripe markers, zebrin II/aldolase C (Brochu et al., 1990
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Figure 4. Parasagittal clusters of dysbindin-immunoreactive mossy fiber terminals bear a consistent relationship to the overlying Purkinje cell compartmentation. A, Double-immunofluorescence staining with anti-dysbindin (red) and anti-zebrin II (green) in the lobule V of the cerebellar vermis. The Purkinje cell stripes P1+ and P2+ are labeled: the midline cluster of dysbindin-immunoreactive terminals (1) in the granular layer is centered beneath P1+ and the first lateral cluster (2) beneath P2+. B, Because both anti-dysbindin and anti-HSP25 were raised in rabbit, serial 40 µm transverse sections through the NZ (lobules IX and X) were each immunostained with a single antibody and the digital images overlaid (dysbindin, red; HSP25, green). The midline HSP25-immunoreactive stripe is shown (for details of constitutive HSP25 expression in the mouse NZ, see Armstrong et al., 2000
Granular layer heterogeneity: dysbindin immunoreactivity is associated with synapses on unipolar brush cells
Previous studies of mossy fiber chemical heterogeneity in the cerebellum suggest two plausible identities for some of the dysbindin-immunoreactive glomeruli: the mossy fiber projections immunoreactive for choline acetyltransferase (ChAT) (Barmack et al., 1992aTransverse sections through the adult mouse cerebellum were double-immunofluorescence-stained for dysbindin and ChAT (Fig. 5A). Large ChAT-immunoreactive mossy fiber rosettes in the granule cell layer are restricted primarily, but not exclusively, to the NZ (Fig. 5A). A similar distribution was described previously in rabbits (Barmack et al., 1992a
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Figure 5. Heterogeneity of the granular layer. A, transverse section through lobule X double-immunofluorescence-labeled for dysbindin (red) and ChAT (green). The first lateral cluster is shown (see Figs. 2C, 3B). Double-labeled profiles are rare, implying that mossy fiber terminals immunoreactive for ChAT constitute a separate population from those that stain with anti-dysbindin. B, Calretinin immunoreactivity reveals parasagittally oriented clusters of unipolar brush cells. A high-power view of a single unipolar brush cell is shown in the inset. C, High-magnification view of the granular layer showing profiles immunoreactive for calretinin alone, and for both calretinin and dysbindin. D, Confocal image of a single unipolar brush cell suggests that profiles immunoreactive for calretinin alone are primarily unipolar brush cell somata, and profiles immunoreactive for both calretinin and dysbindin are primarily unipolar brush cell dendrites (arrow) associated tightly with mossy fiber terminals (asterisk). E, F, G, Transverse sections through the vermis double-immunofluorescence-stained for nNOS (green) and dysbindin (red). The expression patterns of dysbindin and nNOS are primarily complementary. nNOS immunoreactivity in lobule IV is homogeneous in the molecular layer but highly heterogeneous in the granular layer, in which it is preferentially found in the deeper half, closer to the white matter, in which it is restricted to a complex array of patches. In contrast, dysbindin immunoreactivity is heaviest nearer the Purkinje cell layer. Scale bars: A, 50 µm; B, 250 µm (inset, 50 µm); C, 50 µm; D, 10 µm; E, 250 µm; F, 100 µm; G, 50 µm.
Unipolar brush cells can be identified by using anti-calretinin immunocytochemistry (Dino et al., 1999
Finally, nNOS histochemistry has revealed a complex heterogeneity in the granular layer (Yan et al., 1993
Dysbindin immunoreactivity in Purkinje cell dendrites does not require climbing fiber input
In addition to strong dysbindin immunoreactivity in a subset of mossy fiber synaptic glomeruli, the Purkinje cell somata are usually lightly stained with anti-dysbindin and weak, patchy staining of the cerebellar molecular layer, in particular on the primary shafts of Purkinje cell dendrites, is also seen in most regions of the cerebellar cortex (Fig. 6A). Little or no reaction product was deposited in the secondary dendrites or dendritic spines. If dysbindin is associated with synapses on Purkinje cells then, based on its localization to the smooth primary dendrites, it seems likely that either it is located presynaptically in the climbing fiber terminals or postsynaptically in the dendritic shafts. To differentiate between these possibilities, a single dose of 3-acetylpyridine was used to cause the degeneration of the inferior olivary complex, and thus the climbing fiber afferent projection [rat (Desclin and Colin, 1980
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Figure 6. A, Dysbindin immunoreactivity is associated with Purkinje cell dendrites. Transverse section through the cerebellar vermis immunoperoxidase stained with anti-dysbindin. Reaction product is deposited in mossy fiber synaptic glomeruli in the granular layer, lightly in the Purkinje cell somata in the Purkinje cell layer, and intensely as punctae along the Purkinje cell primary dendritic shafts and their initial branch points in the molecular layer (arrow). There is little or no immunoreactivity in the secondary and tertiary dendrites or the dendritic spines. B, C, Climbing fiber afferents are not necessary to maintain dysbindin expression in the molecular layer. Cresyl violet-stained sections through the medulla oblongata of control mice (B) showing the inferior olivary complex (IOC) and a 3-acetylpyridine-treated animal (C) 11 d after injection showing the almost complete ablation of the inferior olivary complex. D, Confocal microscopic analysis of a section from a 3-acetylpyridine-treated mouse double-immunofluorescence-stained for the Purkinje-cell-specific protein calbindin (green) and dysbindin (red). Purkinje cell bodies and primary dendrites remain immunoreactive for both antigens (arrow), despite the ablation of climbing fiber innervation. Scale bars: A, 100 µm; B, C (in C), 250 µm; D, 20 µm.
Dysbindin expression in the cerebellum of mdx mice
In brain, dysbindin is associated with the dystrobrevins as part of a dystrophin-associated protein complex (Blake et al., 1999The loss of well-defined stripes of dysbindin-immunoreactive mossy fiber terminals in the NZ of mdx mice does not extend to other aspects of cerebellar compartmentation. First, the expression of zebrin II in mdx mice is entirely normal (compare Figs. 8A and 8B). Thus, the absence of dystrophin from the Purkinje cell dendrites does not affect the development of Purkinje cell compartmentation, and the ectopic dysbindin expression by mossy fiber glomeruli is not secondary to abnormal Purkinje cell compartmentation. The same is true for nNOS expression (data not shown). Secondly, the distribution of calretinin immunoreactivity is unchanged (compare Figs. 8C and 8D): parasagittal clusters of calretinin immunoreactive unipolar brush cells are clear in the mdx cerebellum, although glomeruli enriched with dysbindin are homogeneously distributed (and dysbindin-immunoreactive projections by far outnumber the available unipolar brush cell targets in mdx). Thus, ectopic dysbindin expression in mossy fiber glomeruli is not secondary to ectopia of the unipolar brush cells. To address the question of whether the additional anti-dysbindin immunoreactive mossy fiber terminals in mdx reflect a general increase in the number of synapses, we have used peroxidase immunocytochemistry to compare several other proteins associated with mossy fiber presynaptic terminals: ChAT (Barmack et al., 1992a
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Figure 8. Many aspects of cerebellar patterning are unaffected in the mdx mouse. A, C, E, G, I, K, +/+; B, D, F, H, J, L, mdx. A, B, Transverse sections through lobule III (AZ) immunofluorescence stained for zebrin II. The P1+ and P2+ stripes are distinct in both. C,D, Transverse section through lobule IX immunofluorescence stained for calretinin. Roughly equal numbers of reactive profiles (predominantly unipolar brush cells) are stained in both. E, F, Transverse section through lobule IX immunoperoxidase-stained for ChAT. The number of mossy fiber glomeruli immunoreactive for ChAT in mdx mice is unchanged. Three proteins associated with mossy fiber presynaptic terminals were also compared in immunoperoxidase-stained transverse sections through lobule IX: synaptophysin (G, H), synaptoporin (I, J), and cysteine string protein (K, L). No systematic differences were seen between +/+ and mdx. Scale bars: A, B (in B), 500 µm; C, D (in D), 250 µm; E, F (in F), 100 µm; G-L (in H), 50 µm.
Discussion
The parasagittal mossy fiber terminal fields revealed by anti-dysbindin staining are consistent with much that is known about cerebellar anatomy. The striped organization of the cerebellum is best understood from the perspective of Purkinje cell heterogeneity (Herrup and Kuemerle, 1997The distribution of dysbindin-immunoreactive mossy fiber glomeruli in the NZ of the vermis resembles both ChAT immunoreactivity and the distribution of unipolar brush cells (Dino et al., 1999
Dysbindin binds both
Abnormal anti-dysbindin immunostaining in mdx may reflect either mossy fiber sprouting, ectopic dysbindin expression by other mossy fibers, or the unmasking of normally concealed epitopes. Mossy fiber compartmentation is established early in development through the direct interaction between afferent growth cones and embryonic Purkinje cell clusters (for review, see Sotelo and Wassef, 1991
In muscle, the absence of dystrophin in mdx tissue results in the upregulation of dysbindin expression (Benson et al., 2001
Footnotes
Received Feb. 26, 2003; revised Apr. 29, 2003; accepted May. 28, 2003.This work was supported by grants from the Canadian Institutes of Health Research (R.H.), Canadian Institutes of Health Research Training Program in Genetics, Child Development and Health (R.V.S.), and the Wellcome Trust (D.J.B.). D.J.B. is a Wellcome Trust Senior Fellow. We thank Dr. Sarah McFarlane for advice and Dr. Jan Braun for antibodies.
Correspondence should be addressed Dr. R. Hawkes, Department of Cell Biology and Anatomy, Faculty of Medicine, University of Calgary, 3330 Hospital Drive Northwest, Calgary, Alberta T2N 4N1, Canada. E-mail: rhawkes{at}ucalgary.ca.
Copyright © 2003 Society for Neuroscience 0270-6474/03/236576-10$15.00/0
References
Ahn AH, Dziennis S, Hawkes R, Herrup K (1994) The cloning of zebrin II reveals its identity with aldolase C. Development 120: 2081-2090.[Abstract]
Akintunde A, Eisenman LM (1994) External cuneocerebellar projections and Purkinje cell zebrin II bands: a direct comparison of parasagittal banding in the mouse cerebellum. J Chem Neuroanat 7: 75-86.[Web of Science][Medline]
Anderson JL, Head SI, Rae C, Morley JW (2002) Brain function in Duchenne muscular dystrophy. Brain 125: 4-13.
[Abstract/Free Full Text] Armstrong CL, Hawkes R (2000) Pattern formation in the cerebellar cortex. Biochem Cell Biol 78: 551-562.[Web of Science][Medline]
Armstrong CL, Krueger AM, Currie RW, Hawkes R (2000) Constitutive expression of the 25 kDa heat shock protein Hsp25 reveals novel parasagittal bands of Purkinje cells in the adult mouse cerebellar cortex. J Comp Neurol 416: 383-397.[Medline]
Armstrong CL, Krueger-Naug AMR, Currie RW, Hawkes R (2001) Expression of heat-shock protein Hsp25 in mouse Purkinje cells during development reveals novel features of cerebellar compartmentation. J Comp Neurol 429: 7-21.[Medline]
Avila MT, Hong E, Thaker GK (2002a) Current progress in schizophrenia research: eye movement abnormalities in schizophrenia: what is the nature of the deficit? J Nerv Ment Dis 190: 479-480.[Medline]
Avila MT, Weiler MA, Lahti AC, Tamminga CA, Thaker GK (2002b) Effects of ketamine on leading saccades during smooth-pursuit eye movements may implicate cerebellar dysfunction in schizophrenia. Am J Psychiatry 159: 1490-1496.
[Abstract/Free Full Text] Barmack NH, Baughman RW, Eckenstein FP (1992a) Cholinergic innervation of the cerebellum of the rat by secondary vestibular afferents. Ann NY Acad Sci 656: 566-579.[Web of Science][Medline]
Barmack NH, Baughman RW, Eckenstein FP (1992b) Cholinergic innervation of the cerebellum of rat, rabbit, cat, and monkey as revealed by choline acetyltransferase activity and immunohistochemistry. J Comp Neurol 317: 233-249.[Medline]
Barmack NH, Baughman RW, Eckenstein FP, Shojaku H (1992c) Secondary vestibular cholinergic projection to the cerebellum of rabbit and rat as revealed by choline acetyltransferase immunohistochemistry, retrograde and orthograde tracers. J Comp Neurol 317: 250-270.[Web of Science][Medline]
Benson MA, Newey SE, Martin-Rendon E, Hawkes R, Blake DJ (2001) Dysbindin, a novel coiled-coil-containing protein that interacts with the dystrobrevins in muscle and brain. J Biol Chem 276: 24232-24241.
[Abstract/Free Full Text] Blake DJ, Kroger S (2000) The neurobiology of Duchenne muscular dystrophy: learning lessons from muscle? Trends Neurosci 23: 92-99.[Web of Science][Medline]
Blake DJ, Nawrotzki R, Loh NY, Gorecki DC, Davies KE (1998) beta-dystrobrevin, a member of the dystrophin-related protein family. Proc Natl Acad Sci USA 95: 241-246.
[Abstract/Free Full Text] Blake DJ, Hawkes R, Benson MA, Beesley PW (1999) Different dystrophin-like complexes are expressed in neurons and glia. J Cell Biol 147: 645-658.
[Abstract/Free Full Text] Blake DJ, Weir A, Newey SE, Davies KE (2002) Function and genetics of dystrophin and dystrophin-related proteins in muscle. Physiol Rev 82: 291-329.
[Abstract/Free Full Text] Boegman RJ, Parent A, Hawkes R (1988) Zonation in the rat cerebellar cortex: patches of high acetylcholinesterase activity in the granular layer are congruent with Purkinje cell compartments. Brain Res 448: 237-251.[Medline]
Braun JE, Scheller RH (1995) Cysteine string protein, a DnaJ family member, is present on diverse secretory vesicles. Neuropharmacology 34: 1361-1369.[Web of Science][Medline]
Brenman JE, Chao DS, Xia H, Aldape K, Bredt DS (1995) Nitric oxide synthase complexed with dystrophin and absent from skeletal muscle sarcolemma in Duchenne muscular dystrophy. Cell 82: 743-752.[Web of Science][Medline]
Brochu G, Maler L, Hawkes R (1990) Zebrin II: A polypeptide antigen expressed selectively by Purkinje cells reveals compartments in rat and fish cerebellum. J Comp Neurol 291: 538-552.[Web of Science][Medline]
Desclin JC (1976) Early terminal degeneration of cerebellar climbing fibers after destruction of the inferior olive in the rat. Synaptic relationships in the molecular layer. Anat Embryol (Berl) 149: 87-112.[Medline]
Desclin JC, Colin F (1980) The olivocerebellar system. II. Some ultrastructural correlates of inferior olive destruction in the rat. Brain Res 187: 29-46.[Web of Science][Medline]
Desmond JE, Gabrieli JD, Wagner AD, Ginier BL, Glover GH (1997) Lobular patterns of cerebellar activation in verbal working-memory and finger-tapping tasks as revealed by functional MRI. J Neurosci 17: 9675-9685.
[Abstract/Free Full Text] Dino MR, Willard FH, Mugnaini E (1999) Distribution of unipolar brush cells and other calretinin immunoreactive components in the mammalian cerebellar cortex. J Neurocytol 28: 99-123.[Web of Science][Medline]
Dino MR, Nunzi MG, Anelli R, Mugnaini E (2000) Unipolar brush cells of the vestibulocerebellum: afferents and targets. Prog Brain Res 124: 123-137.[Medline]
Eisenman LM, Hawkes R (1993) Antigenic compartmentation in the mouse cerebellar cortex: zebrin and HNK-1 reveal a complex, overlapping molecular topography. J Comp Neurol 335: 586-605.[Web of Science][Medline]
Fykse EM, Takei K, Walch-Solimena C, Geppert M, Jahn R, De Camilli P, Sudhof TC (1993) Relative properties and localizations of synaptic vesicle protein isoforms: the case of the synaptophysins. J Neurosci 13: 4997-5007.[Abstract]
Gravel C, Hawkes R (1990) Parasagittal organization of the rat cerebellar cortex: direct comparison of Purkinje cell compartments and the organization of the spinocerebellar projection. J Comp Neurol 291: 79-102.[Web of Science][Medline]
Hawkes R (1997) An anatomical model of cerebellar modules. Prog Brain Res 114: 39-52.[Web of Science][Medline]
Hawkes R, Eisenman L (1997) Stripes and zones: the origins of regionalization of the adult cerebellum. Perspect Dev Neurobiol 5: 95-105.[Web of Science][Medline]
Hawkes R, Turner RW (1994) Compartmentation of NADPH-diaphorase activity in the mouse cerebellar cortex. J Comp Neurol 346: 499-516.[Medline]
Herrup K, Kuemerle B (1997) The compartmentalization of the cerebellum. Annu Rev Neurosci 20: 61-90.[Web of Science][Medline]
Hess DT, Voogd J (1986) Chemoarchitectonic zonation of the monkey cerebellum. Brain Res 369: 383-387.[Web of Science][Medline]
Hinton VJ, De Vivo DC, Nereo NE, Goldstein E, Stern Y (2000) Poor verbal working memory across intellectual level in boys with Duchenne dystrophy. Neurology 54: 2127-2132.
[Abstract/Free Full Text] Ichimiya T, Okubo Y, Suhara T, Sudo Y (2001) Reduced volume of the cerebellar vermis in neuroleptic-naive schizophrenia. Biol Psychiatry 49: 20-27.[Medline]
Jaarsma D, Dino MR, Cozzari C, Mugnaini E (1996) Cerebellar choline acetyltransferase positive mossy fibres and their granule and unipolar brush cell targets: a model for central cholinergic nicotinic neurotransmission. J Neurocytol 25: 829-842.[Web of Science][Medline]
Jaarsma D, Ruigrok TJ, Caffe R, Cozzari C, Levey AI, Mugnaini E, Voogd J (1997) Cholinergic innervation and receptors in the cerebellum. Prog Brain Res 114: 67-96.[Web of Science][Medline]
Ji Z, Hawkes R (1994) Topography of Purkinje cell compartments and mossy fiber terminal fields in lobules II and III of the rat cerebellar cortex: spinocerebellar and cuneocerebellar projections. Neuroscience 61: 935-954.[Web of Science][Medline]
Ji Z, Hawkes R (1995) Developing mossy fiber terminal fields in the rat cerebellar cortex may segregate because of Purkinje cell compartmentation and not competition. J Comp Neurol 359: 197-212.[Web of Science][Medline]
Knuesel I, Bornhauser BC, Zuellig RA, Heller F, Schaub MC, Fritschy JM (2000) Differential expression of utrophin and dystrophin in CNS neurons: an in situ hybridization and immunohistochemical study. J Comp Neurol 422: 594-611.[Web of Science][Medline]
Kohan SA, Pescatori M, Brecha NC, Mastrogiacomo A, Umbach JA, Gundersen CB (1995) Cysteine string protein immunoreactivity in the nervous system and adrenal gland of rat. J Neurosci 15: 6230-6238.[Abstract]
Leclerc N, Beesley PW, Brown I, Colonnier M, Gurd JW, Paladino T, Hawkes R (1989) Synaptophysin expression during synaptogenesis in the rat cerebellar cortex. J Comp Neurol 280: 197-212.[Web of Science][Medline]
Leclerc N, Doré L, Parent A, Hawkes R (1990) The compartmentalization of the monkey and rat cerebellar cortex: zebrin I and cytochrome oxidase. Brain Res 506: 70-78.[Web of Science][Medline]
Lewis RF, Zee DS (1993) Ocular motor disorders associated with cerebellar lesions: pathophysiology and topical localization. Rev Neurol (Paris) 149: 665-677.[Medline]
Lidov HG, Byers TJ, Watkins SC, Kunkel LM (1990) Localization of dystrophin to postsynaptic regions of central nervous system cortical neurons. Nature 348: 725-728.[Medline]
Lui F, Fonda S, Merlini L, Corazza R (2001) Saccadic eye movements are impaired in Duchenne muscular dystrophy. Doc Ophthalmol 103: 219-228.[Medline]
Marani E, Voogd J (1977) An acetylcholinesterase band pattern in the molecular layer of the cat cerebellum. J Anat 124: 335-345.[Medline]
Mugnaini E, Floris A (1994) The unipolar brush cell: a neglected neuron of the mammalian cerebellar cortex. J Comp Neurol 339: 174-180.[Web of Science][Medline]
Mugnaini E, Dino MR, Jaarsma D (1997) The unipolar brush cells of the mammalian cerebellum and cochlear nucleus: cytology and microcircuitry. Prog Brain Res 114: 131-150.[Web of Science][Medline]
Nunzi MG, Mugnaini E (2000) Unipolar brush cell axons form a large system of intrinsic mossy fibers in the postnatal vestibulocerebellum. J Comp Neurol 422: 55-65.[Web of Science][Medline]
Nunzi MG, Shigemoto R, Mugnaini E (2002) Differential expression of calretinin and metabotropic glutamate receptor mGluR1
Oberdick J, Baader SL, Schilling K (1998) From zebra stripes to postal zones: deciphering patterns of gene expression in the cerebellum. Trends Neurosci 21: 383-390.[Web of Science][Medline]
Okugawa G, Sedvall G, Nordstrom M, Andreasen N, Pierson R, Magnotta V, Agartz I (2002) Selective reduction of the posterior superior vermis in men with chronic schizophrenia. Schizophr Res 55: 61-67.[Web of Science][Medline]
Ozol K, Hawkes R (1997) The compartmentation of the granular layer of the cerebellum. Histol Histopathol 12: 171-184.[Web of Science][Medline]
Ozol K, Hayden JM, Oberdick J, Hawkes R (1999) Transverse zones in the vermis of the mouse cerebellum. J Comp Neurol 412: 95-111.[Web of Science][Medline]
Raymond JL (1998) Learning in the oculomotor system: from molecules to behavior. Curr Opin Neurobiol 8: 770-776.[Web of Science][Medline]
Raymond JL, Lisberger SG, Mauk MD (1996) The cerebellum: a neuronal learning machine? Science 272: 1126-1131.[Abstract]
Schilling K, Schmidt HH, Baader SL (1994) Nitric oxide synthase expression reveals compartments of cerebellar granule cells and suggests a role for mossy fibers in their development. Neuroscience 59: 893-903.[Web of Science][Medline]
Schwab SG, Knapp M, Mondabon S, Hallmayer J, Borrmann-Hassenbach M, Albus M, Lerer B, Rietschel M, Trixler M, Maier W, Wildenauer DB (2003) Support for association of schizophrenia with genetic variation in the 6p22.3 gene, dysbindin, in sib-pair families with linkage and in an additional sample of triad families. Am J Hum Genet 72: 185-190.[Web of Science][Medline]
Serapide MF, Cicirata F, Sotelo C, Panto MR, Parenti R (1994) The pontocerebellar projection: longitudinal zonal distribution of fibers from discrete regions of the pontine nuclei to vermal and parafloccular cortices in the rat. Brain Res 644: 175-180.[Medline]
Sotelo C, Wassef M (1991) Cerebellar development: afferent organization and Purkinje cell heterogeneity. Phil Trans R Soc Lond B Biol Sci 331: 307-313.[Web of Science][Medline]
Straub RE, Jiang Y, MacLean CJ, Ma Y, Webb BT, Myakishev MV, Harris-Kerr C, Wormley B, Sadek H, Kadambi B, Cesare AJ, Gibberman A, Wang X, O'Neill FA, Walsh D, Kendler KS (2002) Genetic variation in the 6p22.3 gene DTNBP1, the human ortholog of the mouse dysbindin gene, is associated with schizophrenia. Am J Hum Genet 71: 337-348.[Web of Science][Medline]
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350-4354.
[Abstract/Free Full Text] Ueda H, Kobayashi T, Mitsui K, Tsurugi K, Tsukahara S, Ohno S (1995) Dystrophin localization at presynapse in rat retina revealed by immunoelectron microscopy. Invest Ophthalmol Vis Sci 36: 2318-2322.
[Abstract/Free Full Text] Wiedenmann B, Franke WW (1985) Identification and localization of synaptophysin, an integral membrane glycoprotein of Mr 38,000 characteristic of presynaptic vesicles. Cell 41: 1017-1028.[Web of Science][Medline]
Yan XX, Yen LS, Garey LJ (1993) Parasagittal patches in the granular layer of the developing and adult rat cerebellum as demonstrated by NADPH-diaphorase histochemistry. NeuroReport 4: 1227-1230.[Medline]
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