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R Hawkes

Publications and source records attributed to R Hawkes.

At least 55 records · Page 3Linked to original sources
PubMed1995

Aldolase C/zebrin II and the regionalization of the cerebellum.

The cerebellum is comprised of multiple bands of cells, each with characteristic afferent and efferent projections, and patterns of gene expression. The most studied example of a striped pattern of expression is the antigen recognized by monoclonal antibody antizebrin II. Zebrin II is expressed by subsets of Purkinje cells that form an array of parasagittal bands that extend rostrocaudally throughout the cerebellar cortex, separated by similar bands of Purkinje cells that do not express zebrin II. Recent cloning studies have revealed that the zebrin II antigen is the respiratory isoenzyme aldolase C. This article reviews the cellular and molecular compartmentation of the cerebellum together with the molecular biology of the aldolase C gene, and speculates on possible reasons for a striped pattern of expression.

Brain Chemistry↗
PubMed1994

Compartmentation of NADPH-diaphorase activity in the mouse cerebellar cortex.

The mammalian cerebellum is built around an array of parasagittal bands of Purkinje cells that can be demonstrated by immunocytochemical staining for the differentiation antigen zebrin II. Climbing and mossy fiber afferents also terminate in bands, and the afferent terminal fields and the Purkinje cell bands are aligned. The convergence of mossy and climbing fiber pathways onto the Purkinje cells, which are the sole output of the cerebellar cortex, is a characteristic feature of cerebellar circuitry. Previous studies showed that when both afferent pathways are activated synchronously there develops a long-term depression of synaptic efficacy at the parallel fiber-Purkinje cell synapse. Two second messenger pathways mediate long-term depression: one involves diacylglycerol and protein kinase C, and the other involves nitric oxide that is generated by a nitric oxide synthase. We have studied the distribution of nitric oxide synthase in the adult mouse cerebellum by using nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry. NADPH-diaphorase activity is found mainly in the granule and basket cells. Within the granular layer NADPH-diaphorase activity is expressed nonuniformly by patches of granular cells and synaptic glomeruli. The patches are seen in all lobules, are reproducible from individual to individual, and are topographically ordered with respect to the Purkinje cell compartments as revealed by using anti-zebrin II immunocytochemistry. These data imply that nitric oxide-dependent, long-term depression may only involve a subset of mossy fiber/granule cell projections, and that one role for nitric oxide may be to refine cerebellar receptive fields.

Animals↗
PubMed1994

Functional and antigenic maps in the rat cerebellum: zebrin compartmentation and vibrissal receptive fields in lobule IXa.

The mammalian cerebellum is compartmentalized, both structurally and biochemically, into an array of parasagittal bands. In the adult rat, bands can be shown by immunocytochemical staining of a Purkinje cell subset with the monoclonal antibody antizebrin II. In contrast to the bands revealed by the zebrin II distribution, electrophysiological maps of tactile representations show an apparently quite different organization, a patchwork somatotopy of interwoven small receptive fields. We have compared zebrin II compartmentation with the distribution of vibrissal receptive fields in the dorsal face of lobule IXa. Nine adult rats were studied. Zebrin II immunocytochemistry revealed a zebrin II+ band at the midline (P1+) and three others (P2+, P3+, P4+) arrayed laterally, separated by the P1-, P2-, and P3- bands of unstained Purkinje cells. The only significant source of variability was that P3- was sometimes ill defined, making the P3+ and P4+ difficult to distinguish. Electrophysiological recording in the granular layer of lobule IXa identified two reproducible vibrissal receptive fields on each side of the midline (V1 and V2), with a third, more laterally, identified occasionally (V3). When receptive field maps were constructed and aligned with the zebrin II compartment maps from the same individuals, the V1 receptive field was centered on P1-, V2 on P2-, and V3 on P3-. However, the receptive fields typically extended beyond the P- band into the neighboring P+ to each side. Thus there is a simple, reproducible vibrissal receptive field organization in lobule IXa that bears a constant relationship to the Purkinje cell compartmentation revealed by zebrin II immunocytochemistry. The biochemical parcellation of the cortex may serve to organize the afferent and efferent projection topography and thus to align the sensory and motor maps in the cerebellum.

Animals↗
PubMed1994

Topography of Purkinje cell compartments and mossy fiber terminal fields in lobules II and III of the rat cerebellar cortex: spinocerebellar and cuneocerebellar projections.

The cerebellar cortex is histologically uniform by conventional staining techniques, but contains an elaborate topography. In particular, on the efferent side the cerebellar cortex can be subdivided into multiple parasagittal compartments based upon the selective expression by Purkinje cell subsets of various molecules, for example the polypeptide antigens zebrin I and II, and on the afferent side many mossy fibers terminate as parasagittal bands of terminals. The relationships between mossy fiber terminal fields and Purkinje cell compartments are important for a full understanding of cerebellar structure and function. In this study the locations of spino- and cuneocerebellar mossy fiber terminal fields in lobules II and III of the rat cerebellum are compared to the compartmentation of the Purkinje cells as revealed by using zebrin II immunocytochemistry. Wheat germ agglutinin-horseradish peroxidase was injected at three different levels in the spinal cord and in the external cuneate nucleus, and the terminal field distributions in lobules II and III of the cerebellar cortex were compared with the Purkinje cell compartmentation. In the anterior lobe, zebrin II immunocytochemistry reveals three prominent, narrow immunoreactive bands of Purkinje cells, P1+ at the midline and P2+ laterally at each side. These are separated and flanked by wide zebrin- compartments (P1- and P2-). There are also less strongly stained P3+ and P4+ bands more laterally. The spinocerebellar terminals in the granular layer are distributed as parasagittally oriented bands. Projections from the lumbar region of the spinal cord terminate in five bands, one at the midline (L1), a second with its medial border midway across P1- and its lateral border at the P2+/P2- interface (L2), and a third extending laterally from midway across P2-. The lateral edge of L3 may align with the P3+/P3- border. The terminal fields labeled by a tracer injection into the thoracic region give a very similar distribution (T1, T2 and T3). The only systematic difference is in T2, which statistical analysis suggests may be broader than L2. In contrast, anterograde tracer injections into the cervical region label synaptic glomeruli scattered throughout the lobule with much weaker or no evidence of banding. The terminal fields of the cuneocerebellar projection have a complementary distribution to those of thoracic and lumbar spinocerebellar terminals. There are two lateral bands, Cu2 and Cu3. Cu2 lies within the Purkinje cell P1-compartment, abutting L1/T1 medially and L2/T2 laterally. Cu3 lies between L2 and L3 within the P2- Purkinje cell compartment. The medial edge of Cu3 is tightly aligned with the P2+/P2- border.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

The development of molecular compartmentation in the cerebellar cortex.

The cerebellum is subdivided into hundreds of discrete modules defined by their connectivity and molecular signatures. Cerebellar compartmentation arises very early in development through the formation of multiple populations of chemically distinct Purkinje cells that migrate in a coordinated fashion to form parasagittal bands of cells. Different Purkinje cell bands are then innervated by discrete subpopulations of cerebellar afferents. Because of its stereotyped and strikingly beautiful organization the cerebellum is an excellent model in which to explore genetic/epigenetic aspects of pattern formation in the central nervous system.

Animals↗
PubMed1994

The cloning of zebrin II reveals its identity with aldolase C.

The sagittal organization of the mammalian cerebellum can be observed at the anatomical, physiological and biochemical level. Previous screening of monoclonal antibodies produced in our laboratory has identified two intracellular antigens, zebrin I and II, that occur exclusively in adult cerebellar Purkinje cells. As their name suggests, the zebrin antibody staining of the Purkinje cell population is not uniform. Rather, zebrin-positive Purkinje cells are organized in stripes or bands that run from anterior to posterior across most of the cerebellum; interposed between the zebrin-positive cells are bands of Purkinje cells that are zebrin-negative. Comparison of the position of the antigenic bands with the anatomy of afferent projections suggests that the bands are congruent with the basic developmental and functional 'compartments' of the cerebellum. We report the isolation of cDNA clones of the 36 x 10(3) M(r) antigen, zebrin II, by screening of a mouse cerebellum cDNA expression library. Sequence analysis reveals a 98% identity between our clone and the glycolytic isozyme, aldolase C. In order to more rigorously demonstrate the identity of the two proteins, we stained adult cerebellum with an independent monoclonal antibody raised against aldolase C. Anti-aldolase staining occurs in a previously unreported pattern of sagittal bands of Purkinje cells; the pattern is identical to that revealed by the zebrin II monoclonal. Further, in situ hybridization of antisense aldolase C riboprobe shows that the accumulation of zebrin II/aldolase C mRNA corresponds to the pattern of the zebrin antigen in Purkinje cells. Zebrin II/aldolase C gene expression is thus regulated at the level of transcription (or mRNA stability). In light of previous work that has demonstrated the cell-autonomous and developmentally regimented expression of zebrin II, further studies of the regulation of this gene may lead to insights about the determination of cerebellar compartmentation.

Amino Acid Sequence↗
PubMed1993

Antigenic compartmentation in the mouse cerebellar cortex: zebrin and HNK-1 reveal a complex, overlapping molecular topography.

Two monoclonal antibodies--anti-zebrin I and anti-HNK-1--have been used to study the compartmentation of the mouse cerebellar cortex. As in other species, the pattern of localization of the Purkinje cell specific antigen zebrin I is confined to a subset of Purkinje cells that are organized into parasagittal bands. The basic pattern consists of two abutting paramedian bands (P1+) and up to three additional vermal bands on either side (P2(+)-P4+). This pattern is altered in the vermal regions of lobules X and VI-VII where all Purkinje cells are immunoreactive. In the hemisphere there are three additional bands present (P5(+)-P7+) plus two shorter bands in the paravermal area (P4b+ and P5a+) that extend from the paramedian lobule through the lobulus simplex. This pattern is very similar, but perhaps not identical, to that previously described for the rat. These results suggest a common mammalian plan for the expression and localization of zebrin I. By using a monoclonal antibody to an epitope associated with HNK-1, we have now identified a novel pattern of compartmentation in mouse cerebellum. The HNK-1 epitope is expressed most notably on Purkinje cells and Golgi cells. The molecular layer immunoreactivity associated with the Purkinje cell dendrites varies in intensity in a systematic and reproducible fashion. This reveals a novel cerebellar compartmentation that is sometimes complementary, sometimes overlapping, to that revealed by anti-zebrin. As a result, it is now possible to subdivide the cerebellar cortex into a still finer mosaic of antigenic patches and bands than was possible by using zebrins alone.

Animals↗
PubMed1993

Collateral sprouting in the electrosensory lateral line lobe of weakly electric teleosts (gymnotiformes) following ricin ablation.

Sprouted collateral axons were observed in the electrosensory lateral line lobe (ELL) of gymnotiform teleosts (Apteronotus leptorhynchus) following the ablation of the supraorbital branch of the anterior lateral line nerve. Ablation was accomplished by using microinjections of the toxic lectin ricin. Sprouted axons were followed for up to 26 weeks postablation. Ricin exposure severely reduced axonal numbers and the peripheral electroreceptors in the region innervated by these fibers. To visualize sprouted fibers, intact lateral line afferent nerve branches were anterogradely labelled with the neuronal tract tracers horseradish peroxidase or cobalt chloride, or the monoclonal antibody Q26A3. Within the four somatotopically organized ELL segments, sprouted collaterals were first observed two weeks after ricin injection in the medial and centromedial segments, and four weeks postinjection in the centrolateral and lateral segments. Sprouting involved intrasegmental, horizontally directed axons from adjacent nerve branch terminal fields, and mixed intra- and extrasegmental, dorsally directed axons from the ELL deep fiber layer. The sprouting response was robust but variable in its timing, peaking between 6 and 12 weeks. Subsequently, the intrasegmental, horizontally directed fibers were retained but the mixed dorsally directed fibers, including all extrasegmental axons, were retracted. Therefore, this sprouting response appears to consist of a collateral overproduction followed by a selective axonal retraction. In our view, the most likely explanation for this axonal retraction is that the descending inputs from the isthmus and the cerebellum, as well as commissural fibers from the contralateral ELL, maintain established somatotopic relationships by eliminating somatotopically mismatched sprouted collaterals.

Animals↗
PubMed1993

Structural and molecular compartmentation in the cerebellum.

Most descriptions treat the cerebellum as a uniform structure, and the possibility of important regional heterogeneities in either chemistry or physiology is rarely considered. However, it is now clear that such an assumption is inappropriate. Instead, there is substantial evidence that the cerebellum is composed of hundreds of distinct modules, each with a precise pattern of inputs and outputs, and expressing a range of molecular signatures. By screening a monoclonal antibody library against cerebellar polypeptides we have identified antigens--zebrins--that reveal some of the cerebellum's covert heterogeneity. This article reviews some of these findings, relates them to the patterns of afferent connectivity, and considers some possible mechanisms through which the modular organization may arise.

Animals↗
PubMed1992

Novel developmental boundary in the cerebellum revealed by zebrin expression in the lurcher (Lc/+) mutant mouse.

The cerebellar cortex contains at least two classes of Purkinje cells, which are organized into alternating arrays of parasagittal bands. The clearest demonstration of this compartmentation is the pattern of expression of a family of polypeptide antigens, the zebrins, which are expressed selectively by Purkinje cell subsets. Furthermore, anterograde tracing experiments show that the zebrin compartments are closely correlated with both afferent and efferent projection maps. The further subdivision of long parasagittal bands into smaller modules may occur through several different mechanisms, including the intrinsic cerebellar lobulation and the selective distribution of afferent terminal fields. However, while the longitudinal subdivisions are straightforwardly shown, the mediolateral boundaries are more subtle. In this report we describe a novel mediolateral and anteroposterior compartmentation boundary in mice, running across lobule VIII, that is revealed by the consequences of the lurcher (Lc/+) allele for zebrin expression. In normal mice zebrin compartmentation develops in several discrete stages: until postnatal day 5 (PD5) there is no zebrin expression; from PD5-PD7 zebrin is found only in the posterior lobe vermis, with immunoreactive Purkinje cells in lobules X, IX, and VIII but not elsewhere; from PD7-PD12 most Purkinje cells in the vermis become zebrin+; from PD12-PD15 immunoreactivity also appears in the hemispheres so that almost all Purkinje cells now are zebrin+; and finally, from PD15-PD25 zebrin is gradually suppressed in those Purkinje cells that are zebrin- in the adult until the mature pattern of parasagittal compartments is revealed. In the Lc/+ mutant the normal developmental progression is interrupted at around PD7. As a result, the pattern of zebrin expression becomes frozen at that stage when immunoreactive Purkinje cells are confined exclusively to the posterior lobe vermis. A reproducible boundary between expressing and nonexpressing zones runs mediolaterally across the dorsal surface of lobule VIII. Apart from zebrin expression itself, there are no obvious structural correlates of this transition. This mediolateral boundary identifies a developmental unit in the posterior lobe vermis of the cerebellum, and provides further evidence that the cerebellum is a highly heterogeneous structure.

Animals↗
PubMed1992

Zebrin II distinguishes the ampullary organ receptive map from the tuberous organ receptive maps during development in the teleost electrosensory lateral line lobe.

In weakly electric gymnotiform teleosts, monoclonal antibody anti-zebrin II recognizes developing pyramidal cells in the ampullary organ-receptive medial segment of the medullary electrosensory lateral line lobe (ELL) and in the mechanoreceptive nucleus medialis. Developing pyramidal cells in the remaining three tuberous organ-receptive lateral ELL segments are unreactive. These results suggest that certain biochemical features of the ELL ampullary organ-receptive medial segment are more similar to the nucleus medialis than to the tuberous organ-receptive ELL segments, and support the hypothesis that the ampullary system evolved from mechanosensory precursors.

Animals↗
PubMed1992

Compartmentation in mammalian cerebellum: Zebrin II and P-path antibodies define three classes of sagittally organized bands of Purkinje cells.

The respective roles of genetic and epigenetic factors in generation of pattern formation in the vertebrate nervous system are still poorly elucidated. The mammalian cerebellum is subdivided in parasagittal modules defined by anatomical, physiological, and biochemical criteria. Immunostaining of adult mouse cerebellum with two monoclonal antibodies, P-path, which recognizes 9-O-acetylated glycolipids, and Zebrin II, which recognizes a 36-kDa protein, reveals three classes of sagittally organized bands of Purkinje cells: two complementary groups distinctly immunoreactive to one antibody but not the other and a third group that contains double-labeled cells. No Purkinje cells could be detected that were unreactive to either antibody. The specific and reproducible topography of these three classes of Purkinje cells may be related to the compartmentation of the cerebellum into developmental genetic modules.

Animals↗
PubMed1992

Distribution of zebrin II in the gigantocerebellum of the mormyrid fish Gnathonemus petersii compared with other teleosts.

Immunocytochemistry has demonstrated unexpected heterogeneity among cerebellar Purkinje cells. For example, monoclonal antibody Mab anti-zebrin II reveals parasagittal bands of immunoreactive Purkinje cells in the mammalian cerebellum, but reveals a non-sagittal cerebellar compartmentation pattern in goldfish and gymnotiform fish. The present paper investigates the cerebellar compartmentation pattern, as reflected in the zebrin II distribution, in two other teleosts, the electric mormyrid fish Gnathonemus petersii with its large and regularly built gigantocerebellum, and the electrosensory osteoglossomorph teleost Xenomystis nigri, by using light as well as electron microscopic immunohistochemical techniques. Zebrin II is expressed only in Purkinje cells, where it is present in the cytoplasm of all neuronal compartments, including spines, distal and proximal dendrites, the cell body, and the initial part, as well as terminal boutons of the axon. Other types of cerebellar neurons, including the eurydendroid projection neurons, are zebrin II-negative. In Gnathonemus, zebrin II-positive Purkinje cells are present in the large caudolateral part of the valvula, in lobes C2, C3, and C4 of the corpus, and in the anterior as well as the posterior part of the caudal cerebellar lobe. Zebrin II-negative Purkinje cells are present in a continuous region encompassing the rostromedial part of the valvula, the lobus transitorius, lobe C1 and the ventral part of lobe C2, and in a small, lateral zone of the posterior part of the caudal lobe. In Xenomystis, all Purkinje cells, including those in the medial valvula and the posterior part of the caudal lobe, appear to react with mab anti-zebrin II. This more widespread distribution may be due to the presence of a second antigenic polypeptide in this species. On the basis of the present findings, it is concluded that the mormyrid lobus transitorius, lobe C1, and the ventral part of lobe C2 probably belong to the valvula, while the corpus is restricted to the dorsal part of lobe C2, lobe C3, and lobe C4. The functional significance of zebrin II expression for different subsets of teleostean Purkinje cells remains unclear, since comparisons of different teleosts reveal no general correlation with particular afferent or efferent connections, nor with special morphological features such as a dendritic palisade pattern or different arrangements of the Purkinje cell bodies. A comparison between mammals and teleosts suggests that a distinct parasagittal cerebellar zonation in teleosts is absent, and the major part of the teleostean cerebellum may be considered as a single (midsagittal) cerebellar zone, with about the same width as one mammalian parasagittal zone.

Animals↗
PubMed1991

Zebrin II immunoreactivity in the rat and in the weakly electric teleost Eigenmannia (gymnotiformes) reveals three modes of Purkinje cell development.

Monoclonal antibody (mab) anti-zebrin II recognizes a single 36-kD polypeptide in Purkinje cells in the rat and fish cerebellum. In the adult rat, zebrin II+ Purkinje cells form, in each hemicerebellum, seven parasagittal bands interposed by zebrin II- bands. We show that, in rats, immunoreactivity first appears caudally at postnatal day 5 and spreads; all Purkinje cells are labelled by postnatal day 12. Subsequently, immunoreactivity is selectively lost so that by day 18 the adult pattern of zebrin II+/-immunoreactive bands is created. This pattern indicates two types of Purkinje cells according to developmental trajectory, zebrin II-/+/-. In the adult gymnotiform teleost Eigenmannia, Purkinje cells in the corpus cerebelli (CCb), lateral valvula cerebelli (VCbl), and eminentia granularis anterior (EGa) are zebrin II+. Purkinje cells in the eminentia granularis posterior (EGp) and medialis (EGm) and the medial valvula cerebelli (VCbm) are zebrin II-. Zebrin II antigenicity is first present at 6 days postspawning (P6) in the EGa and at P8 in the CCb. In the valvula, labelling does not appear until P29. Immunoreactivity in the CCb, VCBl, and the EGa persists in the adult, whereas in the VCbm Purkinje cells become zebrin II- before reaching adulthood. These developmental histories (zebrin II-/+ and zebrin II-/+/-) correspond to the patterns of Purkinje cell development in mammals. Additionally, Eigenmannia has a third class of Purkinje cells, in the EGp and EGm, that never express zebrin II immunoreactivity, indicating that zebrin II expression is not an obligatory feature of Purkinje cell development in all vertebrates.

Animals↗
PubMed1991

Synaptogenesis in the rat suprachiasmatic nucleus: a light microscopic immunocytochemical survey.

MabQ155, a monoclonal antibody against synaptophysin, has been used to conduct a light microscopic survey of synaptogenesis in the suprachiasmatic nucleus of the perinatal rat. Synaptophysin is an integral component of synaptic vesicle membranes which is expressed in growth cones and growth cone filopodia as well as in mature synapses. With the light microscope, mabQ155 immunoreactivity in growth cones can be distinguished from that in presynaptic terminals on the basis of the size of immunoreactive puncta. The current study presents a qualitative and quantitative analysis of synaptogenesis from the day of birth (P0) to postnatal day 10 (P10). In our quantitative analysis we have used daily intervals during the first postnatal week, distinguished between growth cones and presynaptic terminals, and divided the suprachiasmatic nucleus into sampling regions that are related to the progress of synaptogenesis. Our data demonstrate regional differences in synaptogenesis within the suprachiasmatic nucleus (SCN), document the temporal progression from the penetration of growth cones to the appearance of mature synapses, and provide information about gradients of synaptogenesis in the nucleus during development.

Aging↗
PubMed1991

A monoclonal antibody to conotoxin reveals the distribution of a subset of calcium channels in the rat cerebellar cortex.

Voltage-sensitive calcium channels (VSCC) are a family of ionophores having different electrical and pharmacological properties. The omega-conotoxin GVIA (omega-CgTX) is a specific blocker of one subset of VSCCs. Because of the specificity of this toxin, a monoclonal anti-omega-CgTX antibody was generated against a omega-CgTX-key hole limpet hemocyanin conjugate and used as a specific marker to study VSCC distributions. This mab was shown to recognize omega-CgTX on Western blots and to display omega-CgTX-dependent immunoperoxidase staining of rat cerebellum. Incubation of fresh, unfixed sections of adult rat cerebellum in omega-CgTX followed by light fixation and peroxidase immunocytochemistry with mab anti-omega-CgTX revealed a specific pattern of labelling. All principal classes of cerebellar neurons were immunoreactive, but in general glial cells were not stained. Most interestingly, strong focal immunoreactivity was encountered at branching points of Purkinje cell dendrites. This characteristic staining pattern implies that a subset of VSCC is specifically concentrated in these regions and suggests that these channels may play a role in the functional integration of dendritic signals.

Animals↗