Search PubMed⌕ Search

Biomedical subjects

P Rakic

Publications and source records attributed to P Rakic.

At least 73 records · Page 4Linked to original sources
PubMed1996

Polarity of microtubule assemblies during neuronal cell migration.

The active migration of neurons from their sites of origin to their final destinations requires the unidirectional translocation of the nuclei and somatic cytoplasm within the growing leading processes. To explore the cellular machinery underlying this translocation, we determined the polarity of microtubules situated within the leading and trailing processes of migrating cerebellar granule cells in situ. Our analysis reveals that the newly assembled positive ends of the microtubules in the leading process uniformly face the growing tip, while their disintegrating negative ends face the nucleus. In the trailing process, by contrast, microtubule arrays are of mixed polarity. We suggest that the dynamics of slow polymerization in combination with fast disintegration of oriented microtubules create "push" and "pull" forces that contribute to the piston-like saltatory displacement of the nucleus and cytoplasm within the membrane cylinder of the leading process of the migrating neuron.

Animals↗
PubMed1996

Selective expression of m2 muscarinic receptor in the parvocellular channel of the primate visual cortex.

Visual information in primates is relayed from the dorsal lateral geniculate nucleus to the cerebral cortex by three parallel neuronal channels designated the parvocellular, magnocellular, and interlaminar pathways. Here we report that m2 muscarinic acetylcholine receptor in the macaque monkey visual cortex is selectively associated with synaptic circuits subserving the function of only one of these channels. The m2 receptor protein is enriched both in layer IV axons originating from parvocellular layers of the dorsal lateral geniculate nucleus and in cytochrome oxidase poor interblob compartments in layers II and III, which are linked with the parvocellular pathway. In these compartments, m2 receptors appear to be heteroreceptors, i.e., they are associated predominantly with asymmetric, noncholinergic synapses, suggesting a selective role in the modulation of excitatory neurotransmission through the parvocellular visual channel.

Animals↗
PubMed1996

Role of neuron-glial junctional domain proteins in the maintenance and termination of neuronal migration across the embryonic cerebral wall.

To identify glial membrane proteins that contribute to the process of neuronal migration in the developing brain, we developed a polyclonal antiserum (D4) and a monoclonal antibody (NJPA1: neuron-glial junctional polypeptide antibody) that recognize membrane proteins localized to the plasmalemmal junction between migrating neurons and adjacent radial glial fibers (Cameron and Rakic, 1994). Here, we show that in the developing cerebral cortex, immunoreactivity for these junctional polypeptides is present throughout the neuronal migratory pathway but becomes minimal or absent where radial glial cell processes enter the marginal zone region, the barrier at which newly arrived neurons normally stop their migration and detach from their glial fiber substrate. We thus tested, using imprints of embryonic cerebral wall and slice preparations, whether the junctional membrane proteins detected by our antibodies contribute to the regulation of neuronal migration in the cerebral cortex. The rate of neuronal migration on glial cell substrates was reduced significantly in the presence of D4 or NJPA1 antibodies. Antibody exposure typically led to the withdrawal of leading processes, changes in microtubular organization and, in some instances, to detachment of neurons from their glial cell substrates. These results suggest that the polypeptides recognized by the D4 and NJPA1 antibodies are essential for the maintenance of normal neuronal migration. Dismantling of neuron-glial cell junctional domains formed by these membrane proteins may underlie neuronal cell detachment from glial migratory substrates at the interface between cortical plate and marginal zone in the developing cerebral wall.

Animals↗
PubMed1996

Intracellular Ca2+ fluctuations modulate the rate of neuronal migration.

Transient elevations of intracellular Ca2+ levels play critical roles in neuronal development, but such elevations have not been demonstrated in migrating neurons. Here, we show that the amplitude and frequency components of Ca2+ fluctuations are correlated positively with the rate of granule cell movement in cerebellar microexplant cultures. Moreover, depression of the amplitude and frequency components of Ca2+ fluctuations by blockade of Ca2+ influx across the plasma membrane results in a reversible retardation of cell movement. These results indicate that the combination of amplitude and frequency components of intracellular Ca2+ fluctuations may provide an intracellular signal controlling the rate of neuronal cell migration.

Animals↗
PubMed1996

Synaptogenesis in the occipital cortex of macaque monkey devoid of retinal input from early embryonic stages.

The role of input from the retina on the development of synaptic organization in the primate striate cortex was examined in macaque monkeys enucleated at embryonic (E) day 67 and E59. Both the prenatally operated animals and their age-matched controls were delivered at term (E165) and killed either at 3 months (at the end of the rapid phase of synaptogenesis) or 3 years (at the end of the plateau phase of synaptogenesis). As expected, in the operated animals the striate cortex had a smaller surface area but a normal thickness and complement of layers. The present study revealed that the mean densities of synaptic contacts per unit area and volume of neuropil in the striate cortex of the two operated animals were similar to those of age-matched controls (approximately 30/100 microm2 or 100/100 microm3 of neuropil). Thus, the absence of retinal input via the lateral geniculate nucleus did not affect the schedule and magnitude of synaptogenesis. Likewise, the ratio of symmetrical versus asymmetrical synapses and mean lengths of synaptic junctions were within the normal range of variation in both group of animals. The proportions of synaptic contacts situated on dendritic spines and shafts were also similar in supra- and infragranular cortical layers of normal and enucleated animals. However, the ratio of synapses situated on dendritic spines and shafts in the sublayers IVAB and IVC, which normally become reversed during late adolescence, were not reversed in the enucleates. Therefore, our study indicated that certain parameters of synaptic development, such as the density of contacts per unit volume of neuropil and the proportion of basic types and their size, in the supra- and infragranular layers of the striate cortex develop to an optimal normal level in the absence of both retinae from early embryonic stages. However, in the thalamorecipient sublayers the details of the synaptic circuits such as their localization on dendritic spines or shafts, fail to mature properly in the absence of normal functional input from the periphery.

Animals↗
PubMed1996

Spatiotemporal gradients of cell genesis in the primate retina.

A cardinal event in the development of all brain structures is the time at which progenitor cells leave the cell cycle and begin to differentiate. We examined cell genesis in the retina of the macaque monkey (Macaca mulatta) by labeling dividing cells with radioactive thymidine ([3H]TdR) and following their fate at terminal division by virtue of their remaining radiolabeled after a long survival period. A number of distinct patterns of cell genesis were observed. The two tissues generated by the optic vesicle, the retinal pigment epithelium and neuroretina, share closely coincident temporal and spatial patterns of cell genesis, indicating that this process may be controlled by a common mechanism. Although overlapping to varying degrees, a clear sequence of genesis was revealed between specific cell types within the neuroretina: ganglion cells are generated first, followed by horizontal cells, cone photoreceptors, amacrine cells, Müller cells, bipolar cells, and, finally, rod photoreceptors. Retinal ganglion cells of differing soma diameter are born at different times-the smallest cells are generated early, the largest late, suggesting a further refined sequence of the functional classes of monkey retinal ganglion cells (first P gamma, then P beta, last P alpha). In addition, at sites where a homogeneous population of cells are crowded and stacked on top of each other (the foveola and perifovea for cones and ganglion cells, respectively) there is a vitreal-to-scleral intralaminar pattern of [3H]TdR labeled cell placement, which reflects both time of genesis and pattern of movement during foveation. These gradients suggest several scenarios for cell fate specification in the retina, many of which might not be obvious in mammals that develop more quickly and have less specialized retinal structure. Thus, data from the highly specialized and slowly developing macaque retina can help to understand visual development in humans and indicate useful avenues for future experimental studies in other species.

Animals↗
PubMed1996

Development of photoreceptor mosaics in the primate retina.

The mosaic of rods and cones in the primate retina is the neuronal array where the visual world is first mapped onto the central nervous system. Rods, which mediate scotopic vision, and cones, which mediate photopic and color vision, are found in all vertebrate species. However, regional differences in the topographic arrangement and ratio of rods to cones vary dramatically among species, including different primates. Furthermore, the proportion and distribution of the wavelength-sensitive cone subtypes vary considerably between primates that occupy different visual habitats. What genetic or environmentally regulated mechanisms specify the position, phenotype, and ratios of photoreceptor subtypes? Available data suggest that the transient appearance of early-differentiating cones may serve to delineate basic species-specific retinal coordinates and determine the opsin phenotype of local assemblies of cones in the fetal photoreceptor mosaic. This article will summarize presently available data and our ideas of how the photoreceptor mosaic is organized in the adult primate retina, the features of these mosaics, which vary between primate species, and the developmental mechanisms, which may account for the emergence of photoreceptor position and specification of their phenotypes in the primate retina.

Animals↗
PubMed1995

Genesis of the retinal pigment epithelium in the macaque monkey.

The development of the retinal pigment epithelium (RPE) was studied in rhesus monkey (Macaca mulatta) fetuses, neonates, and juveniles exposed to a pulse of 3H-thymidine (3H-TdR) between embryonic day (E) 25 and postnatal day (P) 204 and examined at short and long intervals after the injection of the isotope. The RPE develops from the outer layer of the optic cup which by E45 consists of a multistratified epithelium. The outer layer appears immature near the retina's edge and gradually becomes monostratified and more mature centrally. Even at this early stage, all cells contain pigmented melanosomes, although peripherally the pigment is limited to the apical portion of the cells. Examination of autoradiograms from animals allowed to survive for several postnatal months shows that monkey RPE cell genesis begins just after E27, increasing to a peak frequency of 0.38 cells/mm at E43. Between E30 and E85 the density of radiolabelled cells varies within a restricted range of from 0.2 to 0.4 cells/mm (mean = 0.25 +/- 0.09). From the density of radiolabelled cells, and data on the overall density of RPE cells in the juvenile retina, we determined the labelling index. During the first half of gestation, between 0.38% and 0.99% (mean = 0.65 +/- 0.22) of RPE cells are generated during the short interval of isotope availability after pulse injection. Approximately 5% of RPE cells were generated by E33, and 50% by E71. After E85, RPE cytogenesis begins gradually to decrease, and 95% of the cells have been generated by the time of birth. Continued, very low density (0.01 cells/mm) cytogenesis in the RPE is seen at P17, and persists at least until seven months postnatally. RPE cell genesis begins near the fovea, and proceeds towards the periphery. Cell division largely ceases in both foveal and perifoveal regions by E56, at which time labelled cells first begin to appear peripheral to the equator. Besides the timing differences, RPE genesis in the central retina differs from that in the peripheral retina in that it proceeds at a higher rate, and lasts for a shorter time period. A prolonged postnatal period of low density RPE cell genesis persists in both central and peripheral retina. Comparison of the pattern of expansion of the area containing radiolabelled cells in the RPE and neuroretina demonstrates a remarkable spatial and temporal correspondence. Close analysis suggests that at any point on the retina, the last cells are generated in the neuroretina slightly before the last cells in the RPE.

Animals↗
PubMed1995

Neurotransmitter receptors in the proliferative zones of the developing primate occipital lobe.

Film autoradiography was used to investigate the expression of several neurotransmitter receptor subtypes in the transient ventricular and subventricular proliferative zones of the developing occipital lobe in two groups of macaque monkey fetuses. The first group of fetuses were between 60 and 93 days after conception (E60-E93), when the ventricular and subventricular zones of the monkey occipital lobe produce neurons destined for the visual cortex. In the second group, fetuses were between E107 and E128, after generation of cortical neurons has ceased. In the E60-E93 group of fetuses, ventricular and subventricular zones displayed high densities of 5-HT1-serotonergic, D1-dopaminergic, alpha 1- and alpha 2-adrenergic and high affinity kainate receptors. The activation of these receptors has previously been shown to stimulate cell proliferation in other cell systems. The possible involvement of these receptors in regulation of neuronal production is also supported by their absence in the deep laminae of the embryonic cerebral wall after E107, after cortical neurogenesis has been completed. The only exception is a high density of alpha 2-adrenergic receptors maintained near the ventricular surface long after all cortical neurons have been generated. We also found that during neurogenesis, proliferative zones in E66-E90 fetuses displayed virtually no 5-HT2-serotonergic, D2-dopaminergic, beta-adrenergic, M1-muscarinic cholinergic, gamma aminobutyric acid (GABA)A, N-methyl-D-aspartate (NMDA), or alpha-amino-3-hydroxy-5-menthy-4-isoxazole proprionate (AMPA) sites; most of these receptor subtypes have been reported to mediate the suppression of cell proliferation. The present findings suggest that dividing and/or newly generated cortical neurons are capable of receiving specific signals from multiple neurotransmitters present in their environment.

Animals↗
PubMed1995

Tempo of neurogenesis and synaptogenesis in the primate cingulate mesocortex: comparison with the neocortex.

In the neocortex, the onset of the rapid phase (phase 3) of synaptogenesis occurs after the end of neurogenesis. However, we still do not know whether or not these two developmental events are causally related. The present study compares the time-course and tempo of neurogenesis and synaptogenesis in the anterior cingulate cortex (area 24 of Brodmann) and in the primary visual cortex (area 17) in a series of pre- and postnatal rhesus monkeys. Autoradiographic analysis of animals fetally injected with 3H-thymidine showed that all neurons destined for area 24 are generated by embryonic day 70, which is 30 days earlier than in area 17. The rapid phase of synaptogenesis in area 24 starts during the third embryonic month and continues at the same rate through the remainder of gestation and the first 2 months after birth, as has been seen in neocortical areas examined previously. Statistical analysis of the linear portions of the rapid phase indicates that, although neurogenesis in area 24 is completed 1 month earlier than in area 17, the rapid phase of synaptogenesis occurs 41 days later. Moreover, the tempo of synaptic accretion was remarkably similar to that in motor, somatosensory, visual, or associational areas. All were grouped within the same time window of about 40 days, centered at birth. After the second postnatal month, synaptic density in area 24 remains at a high level until sexual maturity. This work shows that the rapid phase of synaptogenesis in the cingulate mesocortex is not linked temporally to the end of neurogenesis. We suggest that it is regulated by the same genetic or humoral factors that control synaptogenesis in the phylogenetically newer neocortical areas.

Animals↗
PubMed1995

The role of receptor/channel activity in neuronal cell migration.

Confocal laser microscopy, in conjunction with carbocyanine dyes and calcium-sensitive fluorescent indicators, was used in slices and explant cultures of developing cerebellum to study cellular mechanisms underlying a motility of neuronal cell migration. The results indicate that a combination of voltage- and ligand-activated ion channels cooperatively regulates Ca2+ influx into the migrating cells. We suggest that molecules, present in the local cellular milieu, affect cell motility by activating specific ion channels and second messengers that influence polymerization of stiff and contractile cytoskeletal proteins. This early interaction between postmitotic neurons and surrounding cells controls the rate of their movements, sculpts their shapes, establishes their positions, and, therefore, indirectly determines their identities to prior formation of synaptic connections.

Animals↗
PubMed1995

Structure of the embryonic primate spinal cord at the closure of the first reflex arc.

Early development of the spinal cord was studied in macaque monkey embryos, using light- and electron microscopy, Golgi impregnation and [3H]thymidine radioautography. All neurons engaged in the first reflex arc are generated before E27 in the 165 day gestation period. The earliest-generated cells differentiate into motoneurons while the commissural and association neurons are generated later. Synaptogenesis starts at E27 in the basal plate, and 2 days later in the alar plate. The first synapses appear as symmetrical junctions situated on the neuronal perikarya and proximal dendrites. Closure of the first spinal reflex are is established within 2 days and follows an antidromic pattern as related to the physiological spread of nerve impulses: synapses on motoneuronal somata and primary dendrites in the basal plate appear first and are followed by synapses in the alar plate, between dorsal root axon collaterals and somata of borderline (commissural/association) neurons. Commissural axons grow towards the floor plate, cross the midline and proceed caudo-rostrally, while association fibers remain ipsilateral. The first wave of apoptosis (programmed cell death) thins out dense populations of nerve and glial cells by E30. Some of the early-generated borderine cells that form commissural and association interneurons, seem to play the role of transient target cells and die once the definitive axonal pathways are established. Since transient cells form numerous synapses, deprivation from the afferent impulses is not a likely cause of their elimination. The present results indicate that the initial developmental events, including formation of the first reflex arc in the primate spinal cord, occur considerably earlier in respect to birth than in other mammals, but that the schedule of cellular events and cellular mechanisms seem to be the same.

Animals↗
PubMed1995

A small step for the cell, a giant leap for mankind: a hypothesis of neocortical expansion during evolution.

The more than 1000-fold increase in the cortical surface without a comparable increase in its thickness during mammalian evolution is explained in the context of the radial-unit hypothesis of cortical development. According to the proposed model, cortical expansion is the result of changes in proliferation kinetics that increase the number of radial columnar units without changing the number of neurons within each unit significantly. Thus, mutation of a regulatory gene(s) that controls the timing and ratio of symmetric and asymmetric modes of cell divisions in the proliferative zone, coupled with radial constraints in the distribution of migrating neurons, could create an expanded cortical plate with enhanced capacity for establishing new patterns of connectivity that are validated through natural selection.

Animals↗
PubMed1995

Radial and horizontal deployment of clonally related cells in the primate neocortex: relationship to distinct mitotic lineages.

To analyze cell lineage in the rhesus monkey necorotex, we used recombinant retroviruses to label individual progenitor cells in the ventricular zone (VZ), then determined histochemically the distribution of their progeny during and after the period of cortical neurogenesis. Distribution patterns of labeled cells in the VZ suggested the coexistence of asymmetrically and symmetrically dividing progenitor cells, indicating that both postmitotic and mitotic progeny are produced during cortical neurogenesis. In the cortex, several distinct patterns of clonal distribution were observed and interpreted as follows: clones aligned radially suggested that asymmetrically dividing progenitors generate sequential "siblings" that migrate, in tandem, along a common radial path to the cortex. In contrast, clones oriented horizontally within a single lamina suggested that symmetric divisions produce multiple, laterally displaced progenitors which, in turn, simultaneously generate "cousin" cells that migrate, in concert, to the same cortical layer. Thus, we propose that different mitotic lineages, which coexist in the monkey VZ, produce distinct radial or laminar patterns of clone deployment that foreshadow the cytological organization of the adult neocortex.

Animals↗
PubMed1995

Dynamics of granule cell migration: a confocal microscopic study in acute cerebellar slice preparations.

Real-time examination of Dil-labeled, immature granule cells in cerebellar slice preparations reveals several temporal and cytological aspects of neuronal migration that have not been observed in previous in vivo or in vitro systems. Using confocal microscopy we have obtained evidence that rates of cell movement depend critically on the age of the cerebellum. Although there were considerable variations in the speed of individual cells, the average rate of cell migration increased systematically from 9.6 +/- 3.0 microns/hr in cerebella from 7-d-old mice to 18.0 +/- 2.9 microns/hr in cerebella from 13-d-old mice. Consequently, granule cells traversed the developing molecular layer within a relatively constant time period despite the doubling in width of the molecular layer during the second week of postnatal life. Granule cell movement was characterized by alternations of short stationary phases with movement in a forward or backward direction. The net displacement of a cell depended on the duration and frequency of these phases as well as on the speed of movement. Changes in the relative position of Dil crystals attached to the surface of granule cells suggested the existence of a complex topographical flow of plasma membrane during migration. Although a large portion of the plasma membrane seemed to move in register with the nucleus and surrounding cytoplasm, new membrane appeared to be incorporated primarily at the leading process. However, the pattern of membrane flow at the interface between migrating neurons and Bergmann glial fibers could not be determined, since these sites could not be labeled by Dil crystals. The present results are in harmony with the concept that multiple cellular/molecular mechanisms may be engaged in granule cell migration.

Aging↗