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Biomedical subjects

P G Bhide

Publications and source records attributed to P G Bhide.

35 records · Page 2Linked to original sources
PubMed1997

Temporal and spatial patterns of expression of p35, a regulatory subunit of cyclin-dependent kinase 5, in the nervous system of the mouse.

The protein p35 is a regulatory subunit of cyclin-dependent kinase 5. It has no recognized homology to cyclins but binds to and activates cyclin-dependent kinase 5 directly in the absence of other protein molecules. Cyclin-dependent kinase 5 was initially isolated by homology to the key cell cycle regulator cdc2 kinase and later identified as a neuronal kinase that phosphorylates histone H1, tau or neurofilaments. This kinase is localized in axons of the developing and mature nervous system. To understand the role of p35 as a regulator of cyclin-dependent kinase 5 activity in the CNS, we examined the pattern of expression of p35 mRNA in the nervous system of embryonic, early postnatal and adult mice. In separate experiments, we also examined the spatial distribution of cyclin-dependent kinase 5 mRNA and the activity of cyclin-dependent kinase 5/p35 kinase complex. Postmitotic cells express p35 mRNA immediately after they leave the zones of cell proliferation. It is also expressed in developing axonal tracts in the brain. Cyclin-dependent kinase 5 mRNA is present in postmitotic and in proliferative cells throughout the embryonic central nervous system. During early postnatal period signal for p35 mRNA declines while that for cyclin-dependent kinase 5 mRNA increases throughout the brain. In the adult brain although both p35 and cyclin-dependent kinase 5 mRNAs are expressed at relatively high levels in certain structures associated with the limbic system, considerable differences exist in the patterns of their distribution in other parts of the brain. These data suggest that the p35/cyclin-dependent kinase 5 complex may be associated with early events of neuronal development such as neuronal migration and axonal growth while in the limbic system of the mature brain it may be associated with the maintenance of neuronal plasticity.

Aging↗
PubMed1996

Cell cycle kinetics in the embryonic mouse corpus striatum.

Cells of the corpus striatum arise from the lateral ganglionic eminence of the telencephalic neuroepithelium. In mice, the length of the cell cycle and that of its component phases were estimated, and the interkinetic nuclear migratory pattern was characterized for lateral ganglionic progenitors on embryonic day 11 and 12 to gain insights into striatal cytogenetic process. An S-phase labeling paradigm using bromodeoxyuridine was employed. To assess regional variation in proliferative patterns, rostral, middle and caudal levels of the ganglionic neuroepithelium were examined separately. The pattern of interkinetic nuclear migration and the duration of G1-, G2-, and M-phases at the rostral and middle levels differed from those at the caudal level. The length of the cell cycle and that of the G1-phase increased during the interval embryonic day 11 to 12, especially at the rostral and middle levels. During the same interval, a sizable secondary proliferative population emerged at all three levels. Two principal conclusions are drawn: Progenitors at the different rostrocaudal levels are heterogeneous with regard to the pattern of cellular proliferation, and ganglionic progenitors are in advance of the cerebral cortical progenitors based on the relative size of the secondary proliferative population and the magnitude of cytokinetic parameters. Thus, proliferative behavior distinguishes progenitor populations at different rostrocaudal levels within the lateral ganglionic neuroepithelium and across the ganglionic and cerebral cortical domains of the telencephalic neuroepithelium.

Animals↗
PubMed1996

Expression of normal and mutant huntingtin in the developing brain.

Huntington's disease (HD) is caused by a genetic mutation that results in a polyglutamine expansion in huntingtin. The time course of neuronal loss in the HD striatum and other affected brain regions before the onset of symptoms is unknown. To determine the potential influence of huntingtin on brain development, we examined its expression in the developing mouse and in human control and HD brain. By Western blot, huntingtin was detected throughout the adult mouse brain and at all stages of embryonic and postnatal brain development. The protein increased significantly between postnatal day 7 (P7) and P15, which marks a period of active neuronal differentiation and enhanced sensitivity to excitotoxic injury in the rodent striatum. Immunoreactivity was found in neurons throughout the brain and localized mostly to the somatodendritic cytoplasm and to axons in fiber bundles. Staining was variable in different groups of neurons and within the same cell population. In developing brain, huntingtin was limited primarily to neuronal perikarya. Increased immunoreactivity in large neurons followed the gradient of neurogenesis and appeared in the basal forebrain and brainstem by embryonic days 15-17, in regions of cortex by P0-P1, and in the striatum by P7. In human brain at midgestation (19-21 weeks), huntingtin was detected in all regions. The brain of a 10-week-old infant with the expanded HD allele expressed a higher molecular weight mutant form of huntingtin at levels comparable to those of the wild-type protein. Thus, mutant huntingtin is expressed before neuronal maturation is complete. Results suggest that huntingtin has an important constitutive role in neurons during brain development, that heterogeneity in neuronal expression of the protein is developmentally regulated, and that the intraneuronal distribution of huntingtin increases in parallel with neuronal maturation. The presence of mutant huntingtin in the immature HD brain raises the possibility that neurons may be affected during brain development and possibly in the postnatal period when vulnerability to excitotoxic injury is at its peak.

Animals↗
PubMed1995

Neuronal high-affinity glutamate transport in the rat central nervous system.

EAAC1 is a neuronal and epithelial high affinity glutamate transporter previously cloned from rabbit intestine. Here we report the isolation of EAAC 1 from rat brain* and its expression in the central nervous system based on in situ hybridization. Strong signals were detected in brain, spinal cord and retina. Expression of EAAC1 was particularly strong in pyramidal cells of the cerebral cortex, pyramidal cells of the hippocampus, mitral cells of the olfactory bulb, various thalamic nuclei and cells of certain retinal layers. EAAC1 was also expressed in non-glutamatergic neurons such as GABAergic cerebellar Purkinje cells and alpha-motor neurons of the spinal cord. We propose that EAAC1 is not only involved in the sequestration of glutamate at glutamatergic synapses and in protecting neurons from glutamate excitotoxicity, but also in the cellular metabolism involving glutamate.

Amino Acid Sequence↗
PubMed1994

An immunocytochemical marker for hamster retinal ganglion cells.

We examined the specificity and developmental time course of the labelling of retinal ganglion cells in Syrian hamsters by a monoclonal antibody AB5. In adult hamsters, AB5 selectively labelled somata in the ganglion cell layer, dendrites in the inner plexiform layer and axons in the nerve fibre layer. When retinal ganglion cells were retrogradely labelled with DiI prior to AB5 immunocytochemistry, all of the retrogradely labelled retinal ganglion cells in the ganglion cell layer were AB5 immunoreactive, indicating that AB5 labels all classes of ganglion cell in that layer. In retinae depleted of retinal ganglion cells by neonatal optic nerve transections, AB5 did not label any somata or processes, indicating that AB5 specifically labels retinal ganglion cells. During development, AB5 labelling first appeared as a weak staining of cell bodies in the ganglion cell layer on postnatal day 12 (P12; PO = first 24 h following birth) and acquired the staining pattern seen in the adult by postnatal day 14. From the onset of AB5 immunoreactivity, AB5-labelled somata of varying sizes were present across the entire retinal surface. Although AB5 labelled retinal ganglion cell axons in the nerve fibre layer of the retina it did not label the optic nerve or retinal ganglion cell axons in the brain at any age examined. AB5 labelling was also found to be compatible with bromodeoxyuridine immunocytochemistry and, therefore, useful for determining the time of generation of hamster retinal ganglion cells.

Animals↗
PubMed1994

Determinants of in vivo MR imaging of slow axonal transport.

PURPOSE: To investigate specific surface characteristics of magnetic contrast agents based on a monocrystalline iron oxide nanoparticle (MION) that may determine their uptake and/or transport by axons. MATERIALS AND METHODS: MION were modified to have a range of surface charges or were covalently linked to wheat germ agglutinin (WGA), a neurotropic protein. Each agent was injected directly into the sciatic nerves or femoral arteries of rats (n = 22), and magnetic resonance (MR) images were obtained several days later. The imaging results then were correlated with results at postmortem histologic examination. RESULTS: Substantial uptake and/or transport by axons occurred only after intraneural injection and only if the agent had a strong surface charge or was covalently linked to WGA. The sciatic nerves appeared as uniformly hypointense structures having lengths proportional to the time from injection to imaging, and the calculated transport rates (4-7 mm/d) were consistent with slow axonal transport. Numerous Schwann cells and macrophages acquired large fractions of the injected agents and contributed substantially to the imaging results. CONCLUSION: Those characteristics of MION-based contrast agents that promote efficacy after intraneural injection may impede delivery to the nerve after intraarterial injection.

Animals↗
PubMed1993

MR imaging of slow axonal transport in vivo.

Three magnetopharmaceuticals based on a monocrystalline iron oxide nanocompound (MION) are evaluated as potential contrast agents for demonstrating axonal transport in vivo by magnetic resonance (MR) imaging. One agent has a strong positive charge, one has a strong negative charge, and the third is covalently linked to wheat germ agglutinin, a plant lectin with a high affinity for axon terminals. All three agents were tagged with rhodamine, and fluorescence microscopy was used to determine their fate after administration and to validate the imaging results. Following injection into or near various neural structures in the motor and visual systems of rats, MR images were obtained at multiple times up to 11 days later, and the imaged tissues were processed for subsequent histological examination. Similar results were obtained with all three agents. Axonal transport was not seen by MR imaging or fluorescence microscopy when the agents were injected into the calf muscles, the vitreous of the eye, or the superior colliculus. However, bidirectional axonal transport was shown unequivocally by both methods after injection directly into the site of a focal crush injury to the sciatic nerve. The nerve, which otherwise is isointense with surrounding tissues on MR images, appeared as a uniformly hypointense structure having a length approximately in proportion to the time from injection to imaging. By 11 days, the course of the nerve was traceable from its component roots in the cauda equina to its bifurcation into the tibial and common peroneal nerves in the leg. A transport rate of about 5 mm/day was calculated, which is consistent with the mechanism of slow transport. MION-based magnetopharmaceuticals thus can be used to demonstrate slow axonal transport, and thereby visualize peripheral nerves, in vivo by MR imaging.

Animals↗
PubMed1992

Axon substitution in the reorganization of developing neural connections.

Insights into the mechanisms of normal and pathological neural development may be gained by studying the reorganization of developing neural connections, caused experimentally or by disease. Many reorganized connections are assumed to arise by the anomalous stabilization of transient connections that occur during normal development. We report that, although the retina projects transiently to the somatosensory system in normal developing hamsters, the permanent retinal projections to the somatosensory system that arise as a consequence of early brain lesions are not formed by the stabilization of the normally transient projection. Instead, the transient retinal axons are replaced by retinal axons that do not normally project to the somatosensory system. The distinction between anomalous stabilization and substitution is significant for determining the cellular mechanisms underlying the development of neural connectivity.

Animals↗
PubMed1991

Stages of growth of hamster retinofugal axons: implications for developing axonal pathways with multiple targets.

Neurons in many regions of the CNS (e.g., cortical areas, thalamic nuclei) are heterogeneous with regard to their afferent and efferent connections. Using the hamster retinofugal system as a model, we investigated the mechanisms by which such connectional heterogeneity arises during ontogeny. Retinal ganglion cell axons were labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil) in paraformaldehyde-fixed tissue. The fluorescent label was photoconverted to a diaminobenzidine reaction product. The morphology of the axons, including their trajectories, branching patterns, and growth cones, was studied at the level of the dorsal lateral geniculate nucleus (LGd) from embryonic day 14 to adulthood. In adult hamsters, axons of retinal ganglion cells (RGCs) are spatially segregated at the level of the lateral geniculate nucleus into a superficial optic tract, situated just beneath the pia, and an internal optic tract consisting of fascicles running parallel to the pia within the geniculate. All retinofugal axons project to the midbrain, but only superficial optic tract axons emit collaterals to the LGd. During development, axons in both divisions of the optic tract emit collaterals to the LGd, but by postnatal day 15, collaterals of internal optic tract axons are virtually entirely eliminated, whereas those of superficial optic tract axons have elaborated terminal arbors. Thus, the heterogeneity among different classes of RGCs with respect to their efferent connections emerges by the selective stabilization, by each class, of a unique subset of connections from an initially widespread set shared by all classes. Thalamic collaterals of RGC axons emerge along established axon trunks, not by bifurcation of the growing tip. This occurs after the axons have grown past the thalamus and, presumably, entered their targets in the midbrain. Growth cones at the tips of elongating axon trunks are larger in size and have a more "complex" morphology compared to the growth cones on collaterals. Axons of RGCs develop in 3 morphologically distinct growth states. First, they elongate to their most distant targets in the midbrain. Then, they simultaneously emit unbranched or poorly branched collaterals to multiple targets. Finally, they elaborate terminal arbors in their definitive targets and eliminate their other collaterals. This developmental strategy may be paradigmatic for the formation of long CNS pathways with multiple targets. Furthermore, these data document, at the single-axon level, the steps in the elaboration and withdrawal of transient neuronal projections.

Animals↗
PubMed1988

Dendritic invagination of developing optic tract axons in the hamster.

Between E15 and P4 in the hamster, axons of retinal ganglion cells in the optic tract over the dorsal lateral geniculate nucleus, are invaginated by, and make synaptic contacts with, small processes interpreted as tips or appendages of geniculate dendrites. In some cases a branch-like protrusion emerges from the axon at or close to the invagination. We hypothesize that the invaginations may be part of the mechanism by which retinocollicular axons are induced to branch and establish the retinogeniculate pathway.

Animals↗
PubMed1985

The effects of a 30 day period of environmental diversity on well-fed and previously undernourished rats: neuronal and synaptic measures in the visual cortex (area 17).

Black and white Lister hooded rats were undernourished from the 16th day of gestation until 25 postnatal days of age. These previously undernourished rats and a set of well-fed rats were later subjected to 30 days of environmental diversity, i.e., environmental enrichment or isolation. Two separate experiments were carried out. In experiment 1, the environmental diversity lasted from 85 to 115 days of age and in experiment 2, from 35 to 65 days of age. At the end of the period of environmental diversity, all rats were killed by perfusion with 2% phosphate-buffered glutaraldehyde. Small pieces of tissue from the right visual cortex were embedded in Spurr's resin. For each rat two blocks of resin-embedded tissue were randomly selected, and from these semithin sections (0.5 micron) were cut and stained with toluidine blue. Photomicrographs of cortical layers II and III were taken from these sections and used to estimate the numerical density of neurons. Ultrathin (ca. 70 nm) sections of the same region of the cortex were cut and stained with lead citrate. These sections were used to estimate the synaptic disc diameter and numerical density. Synapse-to-neuron ratios were calculated from the estimates of synaptic and neuronal numerical densities. In experiment 1, well-fed rats raised in enriched environments had a significantly smaller neuronal numerical density and a greater synaptic disc diameter than well-fed rats raised in an impoverished environment. In experiment 2, neither the well-fed nor previously undernourished rats showed significant effects of environmental treatment on any of the features studied. The statistical interaction between nutrition and environment was not significant for any of the features in either experiment.

Analysis of Variance↗
PubMed1984

The effects of a lengthy period of environmental diversity on well-fed and previously undernourished rats. I. Neurons and glial cells.

Male rats undernourished from the 16th day of gestation until 25 postnatal days of age were raised either in enriched (EC) or isolated (IC) environmental conditions between about 35 and 115 days of age. A parallel set of well-fed rats was raised in identical environments. At the end of this period all rats were killed by perfusion with 2% phosphate buffered glutaraldehyde. Body and forebrain weights and forebrain lengths and widths were determined for each animal. The left cerebral hemisphere was embedded in paraffin-wax and serially sectioned. Three of these coronal sections (defined by subcortical landmarks) taken from the occipital cortex region were used for cortical depth measurements. Small pieces of cortical tissue taken from area 17 of the right cerebral hemisphere were embedded in resin and cut to yield 0.5 micrometer thick sections through the entire depth of the cortex. These sections were used to estimate the nuclear diameters and numerical densities of neurons and oligodendrocytes as well as neuronal perikaryal volumes. Two-way analysis of variance tests on data combined from both nutritional groups revealed significant main effects of nutrition on body weight, forebrain weight, forebrain width, and forebrain length X width values. Environment had significant main effects on forebrain weight, forebrain length, forebrain length X width values, and on neuronal numerical density. The interaction between nutrition and environment was not significant for any of the measurements carried out. These results suggest that 80 days of environmental diversity beginning at about 35 days of age can produce morphological changes in the brains of both well-fed rats and rats undernourished during early life.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1984

The effects of a lengthy period of environmental diversity on well-fed and previously undernourished rats. II. Synapse-to-neuron ratios.

Rats were undernourished from the 16th day of gestation until 25 postnatal days of age and then weaned on to an ad libitum diet. Around 35 days of age, 12 previously undernourished male rats were assigned to an enriched environmental condition (EC) and their littermates to an isolated environmental condition (IC). A parallel set of well-fed rats was similarly assigned. After 80 days in these environmental conditions, all rats were killed by perfusion with 2% phosphate buffered glutaraldehyde. Small pieces of tissue containing the entire depth of the right visual cortex were embedded in Spurr's resin. Semithin (0.5 micron) sections were cut from these blocks and stained with toluidine blue. Photomicrographs of these sections were used to estimate the numerical density of neurons in cortical layers II to III. Ultrathin sections (approximately 70 nm) of the same region of cortex were then prepared for electron microscopy. These were used to estimate the mean synaptic disc diameter and synaptic numerical density. From these estimates of neuronal and synaptic numerical density, synapse-to-neuron ratios were calculated. The results of a two-way analysis of variance test revealed that environment had significant effects on neuronal numerical density, mean synaptic disc diameter, and synapse-to-neuron ratios. Neither nutrition nor its interaction with environment had significant effects on any of the parameters analyzed. These results suggest that environmental diversity can produce alterations in certain neuronal and synaptic characteristics in the visual cortex of both well-fed and previously undernourished rats.

Animals↗
PubMed1984

The effects of environmental diversity on well fed and previously undernourished rats: neuronal and glial cell measurements in the visual cortex (area 17).

Black and white hooded Lister rats were undernourished from the sixteenth day of gestation until 25 postnatal days of age. Around 85 days of age, 12 previously undernourished male rats were assigned to an 'enriched environmental condition' and 12 to an 'isolated environmental condition'. Well fed controls were similarly assigned. After 30 days in these environmental conditions all rats were killed by perfusion with 2% buffered glutaraldehyde. Body and forebrain weights and forebrain lengths and widths were determined for each animal. Cortical depths were measured from sections through the left occipital cortical region. Neuronal and glial cell nuclear diameters and numerical densities as well as neuronal perikaryal volumes were determined from sections through the right visual cortex. In both well fed and previously undernourished groups, the environmentally enriched rats had heavier forebrains and greater forebrain lengths compared to isolated rats. There were no significant differences between enriched and isolated rats in forebrain width or cortical depth measurements in either nutritional group. In both the well fed and previously undernourished groups there were no consistently significant differences between enriched and isolated rats in any of the measurements on neurons and glial cells. Two-way analysis of variance tests on combined data from both nutritional groups indicated significant effects of environment on forebrain weight, forebrain length and on cortical depth in one of the three sections studied (section 10). Nutrition had a significant effect on body weight, forebrain weight and forebrain width. The interaction between nutrition and environment was not statistically significant for any of the measurements carried out.

Animals↗
PubMedDate not supplied

Transient axonal branching in the developing corpus callosum.

During development, there is a transient overproduction of axons in the corpus callosum; this overproduction of axons is due, in part, to a transient excess of neurons that send an axon through the corpus callosum. However, transient axonal branching could also contribute to the developmental overproduction of callosal axons. To investigate this possibility, we filled developing callosal axons in the Syrian hamster with the carbocyanine dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil). Light microscopic analysis showed that, indeed, developing callosal axons branch transiently in the hamster: branching was robust on postnatal day 0 (P0) and P3 (P0 = the first 24 hr after birth), less prominent on P6 and P8, and absent by P11. Immature callosal axons branched before or after crossing the midline and at all rostral-caudal and medial-lateral levels within the corpus callosum. The majority of callosal axon collaterals that were contained within individual 100-micron-thick sections were relatively short (mean = 15.1 microns) but some collaterals extended up to approximately 135 microns from the main axon trunk before passing out of the section in which they were observed. Nearly all of the collaterals emanated from the main axon trunk; higher-order collaterals were rare. Some callosal axon trunks had multiple collaterals. Branching callosal axons originated from multiple cortical areas, including area 17. Electron microscopic observations indicated that the processes designated as axon collaterals by light microscopic criteria would have been included in electron microscopic counts of developing callosal axons. Some callosal axon trunks and branches had ultrastructural features that suggested they were degenerating. In cats, developing callosal axons branch on embryonic day 57 (E57; the first 24 hr after conception = E0) and P0. Thus, it is likely that transient branching of immature callosal axons is a generalized feature of mammalian cortical development and that it contributes to the overproduction of callosal axons, albeit perhaps to varying degrees, in multiple species.

Aging↗
PubMedDate not supplied

Effects of environmental diversity on brain morphology.

This article provides a review of the literature on the effects of early environmental enrichment or impoverishment on the morphological structure of the brain. A critical re-appraisal of this literature shows that many of these effects have not been consistently observed in all experiments. In many experiments where effects have been observed the morphological methods adopted have not always been appropriate for the features being studied and thus the conclusions are brought into question. The review has also examined recent research concerned with the question of studying whether environmental enrichment in later life can ameliorate some of the morphological deficits known to be produced by undernutrition during early life. A consistent finding in this research has been that there is no statistically significant interaction between nutrition and environment, even for those few features which showed a main effect of environment. This indicates that undernutrition during early life does not preclude animals from interacting with their environment in a manner which can produce similar changes in brain morphology to those observed in normally nourished animals.

Animals↗