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P Rakic

Publications and source records attributed to P Rakic.

At least 91 records · Page 5Linked to original sources
PubMed1995

Distribution and density of monoamine receptors in the primate visual cortex devoid of retinal input from early embryonic stages.

Developmental mechanisms that regulate the areal and laminar distribution of various macromolecules, including neurotransmitter receptors in the cerebral cortex, are not known. In the present study, we examined the development of monoaminergic receptors in the rhesus monkey striate and peristriate visual cortex in the absence of input from the retina. Binocular enucleation was performed between embryonic days E60 and E81, prior to the ingrowth of geniculocortical fibers into the cortical plate and before genesis of the granular and supragranular layers of the visual cortex. The animals were delivered at term (E165) and sacrificed at 2 or 12 months of age, and their brains frozen and the occipital lobes cut at 20 microns in the coronal plane. Cortical binding of 3H-clonidine, 125I-pindolol, 3H-5-HT, 3H-ketanserin, 3H-spiperone, 3H-SCH23390, and 3H-prazosin that label various monoamine receptors were autoradiographically visualized and quantified using a computer imaging system. All radioligands displayed specific laminar patterns in the striate and prestriate areas in both groups of animals. The areal and laminar distribution in the anophthalmic monkeys was similar to that in the controls. Significantly, in all enucleated animals, just as in the controls, a particularly high density of 3H-clonidine and 3H-prazosin was observed in the sublayers of layer IV involved in color vision. The present results show that the monoamine receptors in primate visual cortex can establish and maintain distinct laminar and areal patterns in the absence of activity or molecular cues originated from the retina, and provide new insight into the cortical consequences of secondary congenital anophthalmia.

Animals↗
PubMed1994

An array of early differentiating cones precedes the emergence of the photoreceptor mosaic in the fetal monkey retina.

We previously have demonstrated that approximately 10% of cones in the fetal monkey retina precociously express the red/green opsin. These data suggested the possibility that a subset of cones differentiates prior to their nascent cone neighbors. To further assess this early cone differentiation in the fetal monkey retina, we used monoclonal antibodies proven to be important developmental markers of photoreceptor phenotypes and synaptogenesis (XAP-1, specific to photoreceptor membranes; SV2, specific to synaptic vesicle protein). Although these two antibodies recognize functionally distinct antigens, our analyses revealed that both identify a subset of precociously immunoreactive cones. Further, XAP-1- and SV2-positive cones are distributed in the same pattern as precocious red/green-sensitive cones in immature regions of the fetal monkey retina. These results support the hypothesis that the primate retina possesses a spatially organized protomap that may induce the emergence of the photoreceptor mosaic and trigger the formation of color-specific pathways that include horizontal, bipolar, and retinal ganglion cells.

Animals↗
PubMed1994

Early neurogenesis in the anterior hypothalamus of the rhesus monkey.

The time of formation of neurons of the various nuclei of the anterior hypothalamus of the rhesus monkey has been determined using [3H]thymidine autoradiography. Plotting of heavily radiolabeled nuclei indicate that in the macaque monkey neurons of the hypothalamus are generated during three weeks, between the 27th and 48th day of the 165 gestational period of this species. The individual nuclei generally display two waves of neuronal generation. Furthermore, a caudolateral to rostromedial gradient of neurogenesis has been observed in the supraoptic nucleus and a dorsal to ventral gradient in the paraventricular nucleus. Thus, neurons of the primate hypothalamus are generated at a much earlier stage of gestation and their generation lasts about three times longer than in rodents. However, most of the basic neurogenetic gradients are remarkably similar, in spite of species specific differences in cytoarchitecture of specific nuclei.

Animals↗
PubMed1994

Axon overproduction and elimination in the anterior commissure of the developing rhesus monkey.

We have analyzed axon overproduction and elimination in the anterior commissure (AC) of 16 fetal, neonatal, and juvenile rhesus monkeys. Axons are added to the AC at an average rate of 115,000/day during the last two-thirds of gestation, and growth cones are present in a constant proportion to AC axons throughout this period. The peak number of approximately 11 million axons in the AC is reached at birth. Thereafter, axons are eliminated at a net rate of approximately 1 axon/sec during the first 3 postnatal months until the adult number of approximately 3.3 +/- 0.5 million axons is reached. Although there is considerable variability in AC axon number during the period of axon loss, the adult number of AC axons is relatively invariant among the eight adult rhesus monkeys examined. Increase in axon diameter and myelination begins before the major phase of axon elimination and is completed long after the adult number of axons is reached. Apparently, myelinated axons are not eliminated from the AC. Quantitative differences in the magnitude and timing of axon overproduction and elimination in the AC versus that in the corpus callosum (LaMantia and Rakic [1990] J. Neurosci. 10:2156) indicate specific modulation of the development of each commissure, perhaps reflecting differences in the developmental history and functional identity of the distinct cortical regions that give rise to them. This process of overproduction and elimination of AC axons during postnatal development in primates might contribute to individual variations in AC size correlated with a wide range of physical and behavioral differences.

Animals↗
PubMed1994

Neurons and astroglia express distinct subsets of Na,K-ATPase alpha and beta subunits.

We have analyzed the expression pattern of Na,K-ATPase alpha and beta subunit isoforms within the rodent and primate central nervous system. Membrane fractions prepared from rat cerebral cortical type-1 astrocytes and rat cerebellar granule and hippocampal neurons were characterized by immunoblot analyses using a panel of alpha and beta subunit isoform-specific antisera. Each cell type was found to express the alpha 1 isoform but showed differences in the expression of other subunits. Cortical astrocytes displayed alpha 2 and beta 2 subunits, whereas cerebellar granule neurons showed expression of alpha 3 and beta 1 subunits. All three alpha subunit isotypes were detected in hippocampal neurons. A survey of the immunofluorescent staining pattern of the alpha 3 subunit in rat and monkey brain confirmed that expression of this Na,K-ATPase alpha subunit isoform was restricted exclusively to neurons. These results suggest that both neurons and astrocytes express multiple, yet distinct, Na,K-ATPase isoenzymes. The identification of cell types expressing limited combinations of alpha and beta subunits should provide a framework for understanding the physiological significance of Na,K-ATPase isoenzyme diversity and may provide useful tools for the analysis of cell lineage in the mammalian central nervous system.

Animals↗
PubMed1994

Recognition, adhesion, transmembrane signaling and cell motility in guided neuronal migration.

Recent studies indicate that migration of neurons from their place of origin to their final destination requires the orchestration of multiple molecular events, including the selection of a pathway by cell recognition receptors, the formation of adhesive interactions with cellular and extracellular substrates through multiple adhesion molecules and the activation of specific ion channels and receptors that provide second messenger mediated signals for the diverse cellular mechanisms involved in cell motility. New approaches allow for the examination of the role of individual molecular components that mediate these processes.

Animals↗
PubMed1994

Unique profiles of the alpha 1-, alpha 2-, and beta-adrenergic receptors in the developing cortical plate and transient embryonic zones of the rhesus monkey.

Film receptor autoradiography was used to study the distribution of alpha 1-, alpha 2-, and beta-adrenergic receptors in the occipital lobe of developing rhesus monkeys ranging in age from the 65th embryonic day (E65) to adulthood (5 years of age). The study shows that each adrenergic receptor subtype has a unique distribution in the cortical plate and transient embryonic zones of the developing cerebral wall. For example, alpha 1 receptors are prominent throughout the proliferative ventricular and subventricular zones while alpha 2 receptors in the same zones form three distinct bands interdigitated with three bands rich in beta sites. There are also considerable temporal differences in the emergence of adrenergic receptor subtypes in specific embryonic zones. For instance, the high density of alpha 1 sites in the germinal zones occurs only in conjunction with intensive proliferative activity. In contrast, beta receptors emerge in these zones after the majority of cortical neurons have been generated. The transient embryonic zones often display higher densities of adrenergic sites than the cortical plate itself. In particular, the subplate zone subjacent to the developing visual cortex contains the highest density of alpha 2-adrenergic sites in the cerebral wall throughout all prenatal ages studies. Finally, the regional differences in the density of beta-adrenergic receptors are evident in the subplate zone underlying the prospective striate and extrastriate cortex before such differences emerge in the cortical plate. The early appearance, unique pattern of distribution, and time-dependent changes of adrenergic receptors in the transient embryonic zones suggest their involvement in the regulation of the cortical development.

Aging↗
PubMed1994

Identification of membrane proteins that comprise the plasmalemmal junction between migrating neurons and radial glial cells.

We have initiated studies to identify membrane polypeptides of radial glial cells that contribute to the selective cell-cell recognition and migration events in developing brain. Of several polyclonal antisera evaluated, one (D4), developed against formaldehyde fixed type 1 cerebellar glial cells, immunolabels the free surface of cortical and cerebellar astroglial and radial glial cells in a patchy pattern. In dissociated glial-neuronal cell cocultures, microdomains of immunoreactivity are detected at the site where the somal region of cells with the morphology of migrating neurons is in contact with an elongated glial cell fiber. Microdomains are absent from oligodendrocytes, process-bearing astrocytes, and neurons. The surface microdomains do not colocalize with components that compose focal adhesion plaques--integrin subunits, vinculin, or actin--and their integrity appears to require an intact microtubule rather than actin cytoskeleton. Furthermore, microdomain structure is maintained in the absence of extracellular Ca and Mg ions. Immunoblot analyses using antibodies affinity purified to individual proteins indicate that the microdomains are composed of two antigens with apparent molecular mass of approximately 48 kDa and approximately 72 kDa. The 48 kDa antigen is not observed in non-neural epithelial tissues and is detected in cortical and cerebellar tissues only at a developmental period that coincides with the stage of active neuronal cell migration. In contrast, the 72 kDa antigen is expressed in many neural and non-neural tissues at late developmental and adult stages. Our data suggest that the identified membrane proteins may contribute to the formation of the junction between migrating neurons and radial glial cell processes and that this junctional complex is linked to the microtubule cytoskeleton.

Aging↗
PubMed1993

Modulation of neuronal migration by NMDA receptors.

The N-methyl-D-aspartate (NMDA) subtype of the glutamate receptor is essential for neuronal differentiation and establishment or elimination of synapses in a developing brain. The activity of the NMDA receptor has now been shown to also regulate the migration of granule cells in slice preparations of the developing mouse cerebellum. First, blockade of NMDA receptors by specific antagonists resulted in the curtailment of cell migration. Second, enhancement of NMDA receptor activity by the removal of magnesium or by the application of glycine increased the rate of cell movement. Third, increase of endogenous extracellular glutamate by inhibition of its uptake accelerated the rate of cell migration. These results suggest that NMDA receptors may play an early role in the regulation of calcium-dependent cell migration before neurons reach their targets and form synaptic contacts.

2-Amino-5-phosphonovalerate↗
PubMed1993

Low-affinity p75 nerve growth factor receptor expression in the embryonic monkey telencephalon: timing and localization in diverse cellular elements.

Monoclonal antibodies against the low-affinity (p75) subunit of the human nerve growth factor receptor have been used to determine the temporal appearance of this receptor and to identify the associated cellular elements in the developing occipital cortex of rhesus monkeys. Adult and fetal brains from embryos at embryonic days 45-121 were used. This embryonic time span includes periods of active neurogenesis, cell migration and initial formation of axonal connections in the cerebral cortex. The first immunolabeling in the developing cerebral wall was seen between embryonic days 56 and 64. The labeling was present in the transient subplate neurons, a small number of axonal processes and pericytes associated with blood vessels. By birth, labeled neurons of the subplate zone disappeared, but immunolabeled axonal processes could now be seen in large numbers in the cortex. These findings are consistent with the role of nerve growth factor in the coordination of cortical differentiation, but not with the initiation of neuronal proliferation, since the emergence of nerve growth factor receptor-labeled elements in the cortex occurs two to three weeks after the onset of neurogenesis in this species. Further, the diverse cellular elements labeled in the fetal cerebrum with the antibodies to the low-affinity nerve growth factor receptor suggests that a receptor or receptors associated with growth factor signaling for more than one growth factor family are recognized by these antibodies. Differential timing in the expression of families of growth factor receptors may be one mechanism by which developing neurons in the cerebral cortex could respond to the different signals which guide such processes as synaptogenesis and morphogenesis.

Animals↗
PubMed1993

Distribution of calcitonin gene-related peptide in vertebrate neuromuscular junctions: relationship to the acetylcholine receptor.

Calcitonin gene-related peptide (CGRP), regarded by several authors to be involved in maintenance of the acetylcholine receptor, is present in the motor axons of various striated rat muscles. It is present, however, only in motor endplates of several selected striated muscles, where it is located in presynaptic axon terminals of neuromuscular junctions. No immunoreactivity could be seen within synaptic vesicles themselves. In the non-human primate Macaca fasciculata, neuromuscular junctions, including those in the diaphragm, display an intense CGRP reaction. The structure of the simian motor endplates is more elaborate than that of the rat. Amphibian motor nerve endings, both in tetanic and tonic muscles, display CGRP immunoreactivity. In tetanic muscles the CGRP reaction outlines "terminaisons en placque" (true motor end plates) and weakly reacting "terminaisons en grappe" (grape-like endings) in tonic muscles. On supramaximal stimulation of the motor nerve, CGRP is depleted from the affected neuromuscular junctions. Wallerian degeneration of the motor axon results in complete disappearance of CGRP. In most rat muscles in which motor endplates do not normally exhibit CGRP immunoreactivity, e.g., the diaphragm and buccinator muscles, the pre-terminal motor axons are CGRP-positive. After immobilization of such muscles by local bupivacaine injection to rats under brief chloral hydrate anesthesia, CGRP immunoreactivity of the neuromuscular junctions can be elicited because blockade of neuromuscular transmission results in accumulation of CGRP in the endplates. Even more striking is the appearance of CGRP immunoreactivity in normally non-reactive motor endplates during axon regeneration after an experimentally induced Wallerian degeneration of the motor axons. We conclude that CGRP is a regular, genotypically determined component of neuromuscular junctions, present either in a manifest or in a latent form. The latter can be elicited by various experimental approaches. The presence of CGRP in the motor endplate supports the theory that this peptide is instrumental in maintenance of the acetylcholine receptor.

Animals↗
PubMed1993

Changes of synaptic density in the primary visual cortex of the macaque monkey from fetal to adult stage.

The kinetics of synaptogenesis in the primary visual cortex (Brodmann's area 17) were analyzed by electron microscopy in 33 rhesus monkeys, ranging in age from the 50th embryonic day (E50) to 20 years. A series of overlapping electron micrographs (vertical probes) were examined at each age on sections of the upper bank of the calcarine fissure. Synaptic contacts were first observed in the E50 specimen in the subplate and marginal zone (prospective layer I). In the cortical plate itself, synapses appear between E65 and E89 starting in the prospective layer VI. By E112, after all cortical neurons have assumed their laminar positions, synapses situated predominantly on dendritic shafts were present at a low density throughout the full thickness of the cortical plate. Thereafter, synapses accumulate more rapidly on dendritic spines and by E144 an equal number of contacts are found on both spines and shafts. The density of synapses continues to increase exponentially in all layers and reaches the mean maximum density of about 90 synapses per 100 microns 3 of neuropil by the third postnatal month. During the next 2 postnatal years the density of synaptic contacts decreases only slightly to a mean of 80/100 microns 3 of neuropil. Around the time of puberty, however, synaptic density decreases more rapidly to reach the adult level of about 40-50/100 microns 3 of neuropil. The 40% decrease in the density of synaptic contacts occurring between 2.7 and 5 years represents a loss of about 5000 synapses per second in the primary visual cortex of the two hemispheres, due primarily to the loss of asymmetric synapses situated on dendritic spines. The transient phase of high density of synaptic contacts located on dendrospines is shorter in thalamo-recipient layer IV than in either supra- or intragranular layers and is completed within the first postnatal year. It ends earlier in sublayer IVC than in layers IVAB and II-III, for example, reflecting biochemical and functional maturation of the different visual subsystems.

Aging↗
PubMed1992

Immunocytochemical localization of DARPP-32, a dopamine and cyclic-AMP-regulated phosphoprotein, in the primate brain.

The localization of DARPP-32, a dopamine and cAMP-regulated phosphoprotein, has been studied in monkey brain by immunocytochemistry. This study indicates that DARPP-32 is enriched in neurons in regions receiving a dense dopamine input from the substantia nigra and ventral tegmental area. Thus, the majority of somata in the anterior olfactory area, nucleus accumbens, caudate nucleus, and putamen are immunoreactive for DARPP-32. In the caudate nucleus, immunoreactive spines receive asymmetric contacts from unlabeled axon terminals. Immunoreactive somata have diameters of 10-15 microns. In regions known to receive projections from these nuclei, immunoreactivity is confined to small puncta that represent axons and axon terminals. Regions in which immunoreactivity is present in puncta include the ventral pallidum, globus pallidus, and substantia nigra pars reticulata. Dopaminergic neurons themselves are not immunoreactive. Neurons containing moderate to weak immunoreactivity for DARPP-32 are observed in portions of the cerebral cortex, particularly in the temporal cortex (layer VI). DARPP-32-positive neurons are also present in the cerebellum, in the medial habenula, and in portions of the bed nucleus of the stria terminalis and amygdaloid complex. DARPP-32 immunoreactivity is also present in astrocytes in the subcortical white matter and in tanycytes in the arcuate nucleus and median eminence. DARPP-32 may be an effective marker for dopaminoceptive neurons in which the actions of dopamine on the D-1 dopamine receptor are mediated through cAMP and its associated protein kinase.

Animals↗
PubMed1992

Genesis of neurons in the retinal ganglion cell layer of the monkey.

We have analyzed the genesis of various neuronal classes and subclasses in the ganglion cell layer of the primate retina. Neurons were classified according to their size and the time of their origin was determined by pulse labeling with 3H-thymidine administered to female monkeys 38 to 70 days pregnant. All offspring were sacrificed postnatally, and their retinas processed for autoradiography. The somata of cells in the retinal ganglion cell layer generated on embryonic day (E) 38 ranged from 9 to 14 microns in diameter. Between E40 and E56, the minimum soma diameter remained around 8-9 microns, while the maximum gradually increased to 22 microns. As a consequence, the means of the distributions of labeled cells also increased with age, from 11.8 microns diameter for cells generated on E38 to 14.6 microns diameter at E56. Over this period the percentage of labeled cells in the 10.5-16.5 microns and greater than 16.5 microns diameter range gradually increased. The proportion of the labeled cells in the less than 10.5 microns diameter range decreased from E38 to E45, but subsequently increased rapidly. At the end of neurogenesis in the retinal ganglion cell layer, around E70, most labeled cells were considerably smaller (7-9 microns) than those generated earlier. Our results indicate that within the ganglion cell layer of the macaque, neurons of small caliber are generated first, followed successively by medium sized cells. Large, putative P alpha cells are generated late. The production between E56 and E70 of cells with the smallest somata suggests that the last-generated neurons in the ganglion cell layer are predominantly displaced amacrine cells. Within the same sector of retina, different classes of neurons in the ganglion cell layer of the rhesus monkey appear to have a sequential schedule of production.

Animals↗
PubMed1992

Selective role of N-type calcium channels in neuronal migration.

Analysis of neuronal migration in mouse cerebellar slice preparations by a laser scanning confocal microscope revealed that postmitotic granule cells initiate their migration only after the expression of N-type calcium channels on their plasmalemmal surface. Furthermore, selective blockade of these channels by addition of omega-conotoxin to the incubation medium curtailed cell movement. In contrast, inhibitors of L- and T-type calcium channels, as well as those of sodium and potassium channels, had no effect on the rate of granule cell migration. These results suggest that N-type calcium channels, which have been predominantly associated with neurotransmitter release in adult brain, also play a transient but specific developmental role in directed migration of immature neurons before the establishment of their synaptic circuits.

Animals↗
PubMed1992

Neurogenesis of the amygdaloid nuclear complex in the rhesus monkey.

The time course of neurogenesis for neurons which comprise the amygdaloid complex in Rhesus monkeys was determined using tritiated thymidine autoradiography. Fourteen pregnant monkeys received injections of tritiated thymidine between embryonic days 27 (E27) and 56 of their 165 day gestation and offspring were sacrificed during the early postnatal period. The first neurons destined for the amygdaloid complex were generated at E33 making them among the earliest postmitotic neurons in the telencephalon. Neurogenesis peaked within all nuclei of the amygdaloid complex between E38 and E48 and had ceased between E50 and E56. While amygdaloid neurogenesis in postnatally sacrificed monkeys displayed a dorsal-to-ventral gradient of radiolabeled neurons, the considerable rotation of the temporal lobe during the latter stages of primate development indicates that neurogenesis in the embryo, during the first third of gestation, actually occurs across a medial-to-lateral gradient. This medial-to-lateral gradient occurs as a smooth wave across the amygdaloid nuclei and does not respect neuroanatomical subdivisions or patterns of connectivity of the amygdaloid nuclei in the Rhesus monkey.

Amygdala↗