PubMed Health⌕ Search

Biomedical subjects

L M Chalupa

Publications and source records attributed to L M Chalupa.

At least 55 records · Page 3Linked to original sources

Process elimination underlies ontogenetic change in the distribution of callosal projection neurons in the postcentral gyrus of the fetal rhesus monkey.

During fetal development, the regional distribution of callosal projection neurons in the rhesus monkey's postcentral gyrus changes from a uniform to a discontinuous pattern. To determine if this developmental change reflects the retraction of transient callosal projections, two different fluorescent tracers were injected into the brain of fetal monkeys of known gestational ages. Fast blue was injected into the entire postcentral gyrus of one hemisphere, whereas a second tracer (rhodamine latex beads or diamidino yellow) was injected into the caudal portion of the postcentral gyrus of the other hemisphere. The rostral portion of the postcentral gyrus (contralateral to the hemisphere injected with fast blue) was subsequently examined for the presence of labeled cells. In animals injected early in fetal development, on embryonic day 110 or younger and sacrificed 4 weeks later, there were numerous cells labeled with both tracers. In contrast, very few double-labeled cells were found in fetuses injected at an older age, embryonic day 135. We interpret these findings as showing that early in fetal development, when callosal projection neurons in the postcentral gyrus show a continuous distribution pattern, single cells in the rostral portion of this gyrus possess at least two collaterals, one projecting to the contralateral hemisphere and the other to the caudal portion of the gyrus. Subsequently, many of these neurons retract callosal collaterals while maintaining ipsilateral projections. Thus, process elimination accounts for the establishment of the discontinuous distribution of callosal neurons found in the postcentral gyrus of the mature primate.

Animals↗

Response properties in the dorsal lateral geniculate nucleus of the adult cat after interruption of prenatal binocular interactions.

1. Single-cell recordings were made in the magnocellular layer of the dorsal lateral genicule nucleus (dLGN) of five adult cats in which prenatal binocular interactions were interrupted by monocular enucleation at known gestational ages. Three cats (early enucleates) had one eye removed on either embryonic day 44.48, or 49, before retinogeniculate inputs are segregated into uniocular layers. Two other (late enucleates) underwent this procedure on embryonic days 55 and 58, when segregation is well advanced. Responses were compared with those obtained from recordings in the A and A1 layers of the dLGN of seven normal adult cats. 2. Cells were classified as ON or OFF by the use of spots of light and as X or Y based on a test of linearity of spatial summation with the use of counterphased sinusoidal gratings. Receptive-field size and spatial resolution were also obtained. 3. The dLGN of prenatally enucleated cats contains a dorsal magnocellular layer and a ventral parvocellular layer. In early enucleates, only an occasional hint of a cell-sparse interlaminar zone was apparent, located between the magnocellular and parvocellular layers. In late enucleates, a prominent cell-sparse band was observed contralateral to the remaining eye, in a region that would most likely correspond to layer A1 in the normal dLGN. No such cell-sparse band was seen ipsilateral to the remaining eye in late enucleates. 4. Eighty-six X cells and 22 Y cells were studied in the enucleates. Both cell types were found at all depths of the magnocellular layer. All but a few neurons had concentric ON-center or OFF-center receptive fields that were normal in size. The topography of receptive fields also appeared normal. In addition, spatial resolution of X and Y cells was similar in experimental and control animals. 5. In early enucleates there was a higher percentage of X cells and a lower percentage of Y cells than normal. The change in X-to-Y ratio was shown to be because of both a gain in cells with X properties and a loss of cells with Y properties. The distribution of dLGN somal sizes in the early enucleates was comparable with controls, so the change in X-to-Y ratio most likely did not result from an electrode sampling bias. It was suggested that the X-to-Y ratio difference could stem from the abnormalities in retinogeniculate terminal arbors that have been shown to follow early eye removal.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Visual receptive fields in the striate-recipient zone of the lateral posterior-pulvinar complex.

The lateral posterior (LP)-pulvinar complex of the cat is known to contain multiple visual areas. In the present study, we examined the receptive field properties of single neurons isolated in the lateral division of this complex (the LPI). The LPI is designated the striate-recipient zone because it is the only region of the LP-pulvinar receiving cortical projections from areas 17 and 18. The recordings revealed that the striate-recipient zone of LP comprises 2 subareas, which we have termed LPI-1 and LPI-2. In the main segment (LPI-1), virtually all cells responded securely to visual stimuli. The vast majority of these neurons were binocular, with relatively small and well-defined receptive fields. More than half of the cells were found to be directionally selective, and almost this many were orientation specific. The orientation tuning of these cells was found to be quite precise, comparable to complex cells in area 17. In contrast, in the small dorsolateral segment of the striate-recipient zone (the LPI-2), a substantial proportion of cells could not be visually activated. Here, the visual cells had very large receptive fields, and relatively few were direction or orientation selective. The LPI-2 receives subcortical inputs from the superficial layers of the superior colliculus, the hypothalamus, and cerebellum, while the LPI-1 is innervated only by cortical axons. It is suggested that the subcortical connections of the LPI-2 account for the differences in the response properties of the 2 striate-recipient areas. The present results, in conjunction with our previous findings on the principal tectorecipient zone (Chalupa et al., 1983), permit 2 generalizations regarding the functional organization of the cat's LP-pulvinar complex. First, there are clear differences among the visual areas of the LP-pulvinar in the cellular processing of visual information. Second, these functional differences can be related to the principal sources of visual input to the various divisions of the LP-pulvinar.

Afferent Pathways↗

Temporal sequence of neurotransmitter expression by developing neurons of fetal monkey visual cortex.

The developing fetal monkey visual cortex was studied immunocytochemically from 110-155 days post-conception in order to localize cell populations immunoreactive (ir) for gamma-aminobutyric acid, Substance P, cholecystokinin-octapeptide, somatostatin, neuropeptide Y, and proenkephalin A peptide (BAM-18). The area 17/18 border and all cortical laminae identified in the adult visual cortex were discernible from the youngest age examined. All ir-cell populations studied were present at each fetal age. However, despite a relatively adult-like cytoarchitecture, all ir-cell populations studied displayed patterns of immunostaining which were unlike those described in adult visual cortex, and showed significant changes in laminar distribution, morphology, and numbers over the time course of gestation examined. Despite the differences in the patterns of immunostaining between the fetal and adult visual cortex, ir-cell populations intrinsic to the developing visual cortex exhibited adult-like combinations of co-localized transmitters and peptides. The developing monkey cortex also contains ir-cell populations, particularly BAM-18-ir cells, which have not been detected immunocytochemically in the adult monkey cortex. Differences between the fetal and the adult ir-cell populations might be accounted for by cell death, morphological transformation, secondary migration or changes in gene expression for neurotransmitters and neuropeptides.

Animals↗

Multiple pathways from the superior colliculus to the extrageniculate visual thalamus of the cat.

The projection from the cat's superior colliculus to the extrageniculate visual thalamus were examined by the anterograde and retrograde transport of WGA-HRP. An acetylthiocholinesterase (ATChE) stain was employed to facilitate the differentiation of regions within the posterior thalamus. On the basis of the distribution of terminal label as well as the laminar origin of projection neurons, four pathways were delineated. Cells in the stratum griseum superficiale (primarily sublaminae II and III) innervate two regions within the nucleus lateralis posterior (LP): the medial zone, which stains darkly for ATChE, and a restricted portion of the lateral zone, adjacent to the pulvinar. Both of these pathways were found to be topographically organized. By using the fluorescent retrograde tracers, fast blue and rhodamine labeled microspheres, it was determined that the inputs to the medial and lateral zones of LP originate primarily from separate cell populations since very few neurons were found to be double-labeled. A third pathway originates principally from cells in the stratum opticum and terminates in an area just below the cholinesterase-rich region of the LP, designated as the ventral division of the LP. The fourth projection is primarily from the stratum griseum intermedium to the suprageniculate complex. Each of these four pathways arises from a population of neurons with heterogeneous morphological characteristics, and for the most part, each pathway comprises morphologically similar cells. These results suggest that visual information from the superior colliculus is conveyed to the extrageniculate thalamus via multiple pathways that may subserve diverse functions.

Acetylcholinesterase↗

Choline acetyltransferase-immunoreactive neurons in fetal monkey cerebral cortex.

Two monoclonal antibodies to choline acetyltransferase (ChAT) were used to stain the cerebral cortex of fetal monkeys at 110-150 days post-conception. In addition to a small number of immunostained fibers, cells resembling typical non-pyramidal neurons were immunostained in developing layers V and VI and in the subjacent white matter of each area examined (sensory-motor and visual areas). ChAT-immunoreactive neurons have not been described in the cerebral cortex of adult primates, but the present observations indicate such neurons exist in the developing primate cortex.

Animals↗

Formation of retinal ganglion cell topography during prenatal development.

A fundamental feature of the mammalian visual system is the nonuniform distribution of ganglion cells across the retinal surface. To understand the ontogenetic processes leading to the formation of retinal ganglion cell topography, changes in the regional density of these neurons were studied in relation to ganglion cell loss and the pattern of retinal growth in the fetal cat. Midway through the gestation period, the density of these neurons was only two to three times greater in the area centralis than in the peripheral retina, whereas shortly before birth this central-to-peripheral difference was nearly 20-fold. Age-related changes in the ganglion cell distribution were found not to correspond in time or magnitude to the massive loss of ganglion cells that occurs during prenatal development. Rather, the formation of ganglion cell density gradients can be accounted for by unequal expansion of the growing fetal retina-peripheral regions expand more than the central region, thereby diluting the peripheral density of ganglion cells to a greater degree. Nonuniform growth, in conjunction with differential periods of neurogenesis of the different types of retinal cells, appears to be a dominant factor regulating overall retinal topography. These results suggest that the differential regional expansion of the fetal retina underlies the formation of magnification factors in the developing visual system.

Animals↗

Retinal crowding alters the morphology of alpha ganglion cells.

It has been hypothesized that the dendritic field size of individual retinal ganglion cells is regulated early in development by interactions among neighboring cells (Wässle and Reiman, Proc. R. Soc. Lond. B. 200:441-461, 1978). An opportunity to test this hypothesis is provided by the consequences of prenatal enucleation that results in an increased density of ganglion cells in the remaining retina (Chalupa et al., Neuroscience 12:1139-1146, 1984). In the present study we compared dendritic field diameters of alpha ganglion cells in normal retinas to those of adult cats that were monocularly enucleated before birth (on embryonic days 42 and 51) and 6 days after birth. In each animal ganglion cells were labeled by central injections of horseradish peroxidase. The retinas were incubated according to a modified Hanker-Yates procedure and whole-mounted. All cells were sampled from the temporal retina along a corridor established by a line drawn through the optic disk and the area centralis. Alpha cells were differentiated into ON and OFF subtypes on the basis of their level of stratification in the inner plexiform layer (Peichl and Wässle, Proc. R. Soc. Lond. B. 212:139-156, 1981). Dendritic field and soma diameters were determined from complete cell drawings by calculating the mean of the two longest orthogonal diameters. Our main findings are: Early monocular enucleation does not disrupt the mosaics of ON and OFF alpha ganglion cells in the remaining retina of adult animals. Within the retinal corridor sampled, the density of alpha cells was substantially greater than normal in the remaining retina of the prenatal enucleates at all eccentricities except the far periphery. Except at the far periphery, the dendritic field diameters of the prenatally enucleated animals were significantly smaller than normal when compared at equivalent eccentricities. However, when compared at equivalent cell densities, dendritic field dimensions in the prenatally enucleated and normal animals were found to be similar. In the prenatal enucleates the somas of these neurons were also significantly smaller than normal. If the monocular enucleation was postnatal, however, the density and the dendritic field diameters of alpha cells did not differ appreciably from normal. These results indicate that the morphology of retinal ganglion cells can be influenced by increasing the density of developing ganglion cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Growth cones, dying axons, and developmental fluctuations in the fiber population of the cat's optic nerve.

We have studied the rise and fall in the number of axons in the optic nerve of fetal and neonatal cats in relation to changes in the ultrastructure of fibers, and in particular, to the characteristics and spatiotemporal distribution of growth cones and necrotic axons. Axons of retinal ganglion cells start to grow through the optic nerve on the 19th day of embryonic development (E-19). As early as E-23 there are 8,000 fibers in the nerve close to the eye. Fibers are added to the nerve at a rate of approximately 50,000 per day from E-28 until E-39--the age at which the peak population of 600,000-700,000 axons is reached. Thereafter, the number decreases rapidly: About 400,000 axons are lost between E-39 and E-53. In contrast, from E-56 until the second week after birth the number of axons decreases at a slow rate. Even as late as postnatal day 12 (P-12) the nerve contains an excess of up to 100,000 fibers. The final number of fibers--140,000-165,000--is reached by the sixth week after birth. Growth cones of retinal ganglion cells are present in the optic nerve from E-19 until E-39. At E-19 and E-23 they have comparatively simple shapes but in older fetuses they are larger and their shapes are more elaborate. As early as E-28 many growth cones have lamellipodia that extend outward from the core region as far as 10 microns. These sheetlike processes are insinuated between bundles of axons and commonly contact 10 to 20 neighboring fibers in single transverse sections. At E-28 growth cones make up 2.0% of the fiber population; at E-33 they make up about 1.0%; from E-36 to E-39 they make up only 0.3% of the population. Virtually none are present in the midorbital part of the nerve on or after E-44. At all ages growth cones are more common at the periphery of the nerve than at its center. This central-to-peripheral gradient increases with age: at E-28 the density of growth cones is two times greater at the edge than at the center but by E-39 the density is four to five times greater. Necrotic fibers are observed as early as E-28 in all parts of the nerve. Their axoplasm is dark and mottled and often contains dense vesiculated structures.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Ontogenetic change in the distribution of callosal projection neurons in the postcentral gyrus of the fetal rhesus monkey.

In the postcentral gyrus of the mature rhesus monkey the distribution of callosal projection neurons is discontinuous. The density of callosal projection neurons, which are mainly located in the supragranular layers, varies both within and across cytoarchitectonic areas (Killackey et al., '83). In the present study, we investigated the ontogeny of corpus callosum projections of the postcentral gyrus in five fetal rhesus monkeys, ranging in age from embryonic day (E) 108 to E 133. Multiple large injections of horseradish peroxidase that involved the underlying white matter were made into the postcentral gyrus of one hemisphere and the distribution of labeled neurons in the ipsilateral thalamus and the other hemisphere was determined. The pattern of thalamic label indicated that the tracer was effectively transported from all portions of the postcentral gyrus. We found that the areal distribution pattern of labeled callosal projection neurons varied at the different fetal ages. At early fetal ages (E 108, E 111, and E 119) callosal projection neurons were continuously distributed throughout the postcentral gyrus. As in the adult animal, the vast majority of labeled callosal projection neurons were found in the supragranular layers, although a few labeled cells were located in the infragranular layers. From the earliest age, there was regional variation in the width of the band of labeled supragranular callosal projection neurons. The difference between the precentral and postcentral gyrus was most obvious, but there was also a difference between anterior and posterior portions of the postcentral gyrus. The first indication of some discontinuity in the distribution of callosal projection neurons was noted at E 126. By E 133, approximately 1 month before birth, the distribution of callosal projection neurons appeared remarkably mature. On E 119 aggregations of anterograde label could be detected in restricted portions of the posterior postcentral gyrus beneath the cortical layers. By E 133 anterograde label was found within the cortical layers (most densely in layer IV) in these regions of the postcentral gyrus. Thus, the emergence of the discrete pattern of callosal projection neurons appears to be temporally correlated with the ingrowth of callosal afferents. On the basis of these observations, as well as those of others (discussed in the text), we propose that the ontogenetic changes in the distribution of callosal projection neurons reflect the unique strategy employed by cortical projection neurons in establishing their patterns of connectivity. It is hypothesized that this strategy may involve multiple processes.

Animals↗

Organization of geniculocortical connections following prenatal interruption of binocular interactions.

The organization of geniculostriate connections in normal cats was compared with that of adult animals that were uniocularly enucleated before birth. In normal animals microelectrophoretic deposits of horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) into the A-lamina of the dorsal lateral geniculate body (LGd) resulted in anterograde label in layers IV and VI and labeled cells in layer VI of areas 17 and 18. The labeling pattern within both of these cortical areas consisted of alternating patches separated by zones of equivalent size that were relatively free of label. In the normal animals no reaction product was evident in any other cortical area. In the prenatally enucleated cats, the LGd both contralateral and ipsilateral to the remaining eye is comprised of only two distinct cell layers. The dorsal layer appears to be a composite of the normal A/A1-laminae, while the ventral layer appears to correspond to the C-laminae. Deposits of WGA-HRP into the superficial aspect of the A/A1 layer yielded a dense continuous band of label within layers IV and VI of areas 17 and 18. Additionally, such deposits in the prenatally enucleated cats also revealed an anomalous reciprocal connection with area 19. Punctate deposits of WGA-HRP into cortical area 19 of the fetal enucleates resulted in the labeling of two distinct populations of cells within the A/A1 layer of the LGd. No cells were labeled within the A-laminae following such deposits into area 19 of normal animals. The geniculocortical connections of the prenatally enucleated cats, including that to area 19, were found to be retinotopically organized. These results indicate that in utero interruption of binocular interactions prevents the formation of ocular dominance domains within areas 17 and 18 of the cat's visual cortex. This could reflect the maintenance of exuberant geniculocortical projections present at the time of prenatal eye removal as originally suggested by Rakic (Science, 214 (1981) 928-931). The anomalous connection with area 19, on the other hand, could be due to the disruption of LGd cell migration resulting from the early eye removal.

Animals↗

Does axonal branching contribute to the overproduction of optic nerve fibers during early development of the cat's visual system?

The number of axons within the optic nerve of the fetal cat was found to be no greater near the chiasm than near the eye. This suggests that the overabundance of axons present in the developing nerve results from a corresponding excess of retinal ganglion cells, and that the elimination of optic fibers that occurs during normal development is principally due to the loss of ganglion cells and not to the elimination of axonal branches.

Animals↗

Selectivity of antibody-mediated destruction of axons in the cat's optic nerve.

The purpose of this study was to determine the selectivity with which polyspecific antibodies directed against large retinal ganglion cells destroy axons in the cat's optic nerve. Immune serum prepared against large ganglion cells isolated from ox retinas was injected into 1 eye of each of 2 cats. After more than 1 month, the cats were perfused with mixed aldehydes and the optic nerves were prepared for transmission electron microscopy. On the basis of a large sample of micrographs of transverse thin sections, we estimated that each of the nerves that issued from treated eyes contained approximately 62,500 necrotic fibers, amounting to 42-46% of the total fiber population in 1 case and 39-42% in the other. The diameters of 2900-5000 intact axons from each of 4 nerves (2 from immune-treated eyes and 2 from untreated eyes) were measured. Comparisons of histograms of axon diameter for nerves from treated and untreated eyes revealed that 90-100% of large axons with diameters above 3.5-4.0 micron were eliminated by the antibodies. Between 65 and 70% of medium-sized fibers were also eliminated. The number of small axons--those with diameters less than 1.2-1.6 micron--did not differ appreciably from normal. These results suggest that the immune serum destroyed virtually all alpha cell axons and a substantial fraction of beta cell axons but did not reduce the number of small fibers that largely stem from the gamma class of retinal ganglion cells.

Animals↗

The laminar distribution of cortical connections with the tecto- and cortico-recipient zones in the cat's lateral posterior nucleus.

The anterograde and retrograde transport of wheat germ agglutinin congugated to horseradish peroxidase was used to examine the laminar organization of cortical connections with the two visual zones that comprise the cat's lateral posterior nucleus. Microelectrophoretic deposits of the tracer into the principal tecto-recipient zone in the medial division of the lateral posterior nucleus revealed reciprocal connections with the following cortical fields: areas 19 and 21a, the medial and lateral banks of the middle suprasylvian sulcus, and the dorsal and ventral banks of the lateral suprasylvian sulcus, which correspond to the dorsal lateral suprasylvian and ventral lateral suprasylvian visual areas of Palmer et al. [(1978) Brain Res. 177, 237-256] and an area in the fundus of the posterior suprasylvian sulcus. In each of these cortical areas two distinct populations of cells were labeled, small pyramidal neurons in layer VI and large pyramidal cells in layer V. Overlying these backfilled cells were two bands of anterograde label, a narrow strip in layer I and a wide band centered in layer IV. Deposits of wheat germ agglutinin conjugated to horseradish peroxidase confined to the striate-recipient zone in the lateral portion of the lateral posterior nucleus resulted in cortical label in areas 17, 18, 19, 20a and b, 21a, the medial and lateral banks of the middle suprasylvian sulcus, the posterior suprasylvian sulcus and in the fundus of the splenial sulcus. In all cortical areas other than 17 and 18, the laminar distribution of label was the same as that found after deposits of the tracer into the medial division of the lateral posterior nucleus. In contrast, areas 17 and 18 contained backfilled cells that were confined to layer V and anterograde label that was restricted to layer I. These findings indicate that the cortical areas that receive a direct projection from the A laminae of the dorsal lateral geniculate nucleus maintain a distinct laminar organization of reciprocal connections with the extrageniculate visual thalamus. Conversely, all other visual areas of the cortex share a common pattern of reciprocal connections with both the tecto- and striate-recipient zones of the lateral posterior nucleus.

Animals↗

Functional organization of the cat's visual cortex after prenatal interruption of binocular interactions.

The functional consequences of interrupting in utero binocular interactions were studied by recording from single cells in area 17 of adult cats that had one eye removed at least 2 wk before birth. In these animals all cortical neurons could be driven by the remaining eye, and in tangential microelectrode penetrations, sequences of neurons containing a full 180-degree cycle of preferred orientations were encountered. Other response properties of cortical neurons in the prenatally enucleated animals were also normal with the notable exception that the dimensions of receptive fields were significantly smaller when compared with those of control animals. Our results indicate that orientation columns in the visual cortex can develop independently of ocular dominance columns, and they suggest that interruption of binocular interactions during prenatal development of the visual pathways may enhance the resolving power of the remaining eye.

Animals↗

An analysis of the transport of WGA-HRP in the cat's visual system.

The increasing use of WGA-HRP as a neuroanatomical tracer has led us to examine two aspects of its uptake and transport. Microelectrophoretic deposits of WGA-HRP were centered in lamina A of the LGd to determine whether or not there is a discrepancy between the size of the deposit site and the effective uptake zone. For this purpose, the size of the deposit site was compared to the area of labeled ganglion cells in the retina. It was found that the effective uptake zone is approximately one half the size of the geniculate area containing reaction-product. Deposits of WGA-HRP in the optic tract, optic radiations and corpus callosum were made in order to determine if this tracer is incorporated and transported by intact fibers. In no case were labeled axon terminals or neuronal somata observed. However, application of the conjugated enzyme to cut fibers of the corpus callosum resulted in robust labeling.

Animals↗

Binocular interaction in the fetal cat regulates the size of the ganglion cell population.

During fetal development of the cat's visual system there is a marked overproliferation of optic nerve axons. In utero binocular interaction contributes to the severity of fiber loss since removal of an eye during gestation attenuates axon loss in the remaining optic nerve. The purpose of the present study was to determine whether this reduced loss of optic nerve fibers is due to a failure of retraction by supernumerary axon branches or to a reduction in ganglion cell death. To resolve this issue, we compared the number of ganglion cells and optic nerve fibers in adult cats which had one eye removed at known gestational ages. Retinal ganglion cells were backfilled with horseradish peroxidase and counts were made from retinal wholemounts. The axon complement was assessed with an electron microscopic assay. In the retinas of a normal cat we estimated 151,000 and 152,000 ganglion cells. The optic nerves of two other normal cats contained approximately 158,000 and 159,000 axons. In comparison, an animal enucleated on embryonic day 42 had 180,000 ganglion cells and 178,000 optic nerve fibers, while in an animal enucleated on embryonic day 51 the corresponding estimates were 182,000 and 190,000. The close agreement between cell and fiber counts indicates that axonal bifurcation does not contribute appreciably to the axon surplus in the optic nerve of prenatally enucleated cats. These results demonstrate that prenatal binocular interaction regulates the size of the mature retinal ganglion cell population.

Animals↗