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L M Chalupa

Publications and source records attributed to L M Chalupa.

At least 37 records · Page 2Linked to original sources
PubMed1996

Topographic organization in the retinocollicular pathway of the fetal cat demonstrated by retrograde labeling of ganglion cells.

The topographic organization of the developing retinocollicular pathway was assessed by making focal deposits of a retrograde tracer (usually rhodamine latex beads) into the superficial layers of the superior colliculus of fetal cats at known gestational ages. Subsequently, the distributions of labeled cells in the contralateral and ipsilateral retinas were examined. At all stages of development, a high density of labeled cells was found in a delimited area (core region) of both retinas. The locations of the retinal regions containing the high density of labeled cells varied with the locus of the tracer deposit in the superior colliculus in a manner consistent with the topographic organization of the mature cat's retinocollicular pathway. Additionally, some labeled ganglion cells, considered to be ectopic, were found to be scattered throughout the contralateral and ipsilateral fetal retinas. Such ectopic cells were few in number throughout prenatal development. For every 100 cells projecting to the appropriate region of the colliculus, we estimate that less than one ganglion cell makes a gross projection error. The incidence of ectopic cells did not differ between the contralateral and ipsilateral retina, even though the overall density of crossed labeled cells was always greater than that of uncrossed labeled cells. In the youngest fetal animals, tracer deposits into the caudal portion of the superior colliculus resulted in a core region of labeled cells in the contralateral nasal retina as well as in the nasal ipsilateral retina. Such uncrossed nasal cells, not seen in more mature animals, appear to innervate the appropriate topographic location of the superior colliculus, but on the wrong side of the brain. Most likely, these uncrossed nasal ganglion cells contribute to the widespread distribution of the ipsilateral retinocollicular pathway observed in fetal cats after intraocular injections of anterograde tracers (Williams and Chalupa, 1982). Collectively, our findings demonstrate that the developing retinocollicular pathway of the fetal cat is characterized by a remarkable degree of topographic precision.

Animals↗
PubMed1996

Specificity of retinal ganglion cell projections in the embryonic rhesus monkey.

Recent studies dealing with the organization of retinal projections in the developing rhesus monkey brain have revealed a high degree of developmental specificity. This is demonstrated by the ingrowth patterns of the initial contingents of crossed and uncrossed fibers that form the primordial optic tract as well as by the adult-like nasotemporal retinal decussation pattern evident even before the period of ganglion cell death. On the basis of these observations, it is suggested that early generated retinal fibers are guided through the optic chiasm by a transiently expressed decussation signal, and that later generated fibers utilize retinal position-dependent cues to innervate the appropriate hemisphere. Furthermore, the first retinal fibers to arrive at the dorsal lateral geniculate nucleus invade only the presumed parvocellular layers. Thus, the initial innervation of the lateral geniculate nucleus appears to reflect the birth order of retinal ganglion cell classes. It is suggested that the high degree of precision evident in the macaque monkey nasotemporal retinal decussation pattern relates to the adultlike distribution of callosal projection neurons in the developing striate cortex of the primate.

Animals↗
PubMed1995

Development of neuropeptide Y immunoreactive amacrine and ganglion cells in the pre- and postnatal cat retina.

In the adult cat, neuropeptide Y (NPY) immunoreactivity (IR) is found within a subgroup of gamma-type ganglion cells and a large group of regularly arrayed amacrine cells. To examine the development of these two cell groups, we charted the appearance and maturation of neuropeptide Y immunoreactivity in the pre- and post-natal cat retina. Neuropeptide Y immunoreactivity is first observed at the central retina within the ganglion cell layer on embryonic day 46, and immunoreactivity within amacrine cells of the inner plexiform layer is present by E50. The number of immunoreactive profiles reaches the adult level in the amacrine population first (around P7), while the ganglion cell population shows a protracted development, with new cells being added until the third postnatal week. NPY-immunoreactive profiles in the ganglion cell layer were confirmed to be ganglion cells by retrograde labeling in both pre- and post-natal animals. Thus, neuropeptide Y-immunoreactive ganglion cells and amacrine cells attain their mature state with very different timecourses, although both cell groups initially follow a central to peripheral pattern of development. Interestingly, NPY expression within the ganglion cell population is temporally correlated with retinal synaptogenesis in the inner plexiform layer. As in the adult cat, NPY-immunoreactive ganglion cells never show a regular distribution during development, while NPY-IR amacrine cells are always distributed regularly even at the earliest ages. The prenatal presence of a regular distribution of NPY-IR amacrine cells suggests that these cells may participate in establishing the ganglion cell mosaics that appear during postnatal development.

Animals↗
PubMed1995

Properties of K+ conductances in cat retinal ganglion cells during the period of activity-mediated refinements in retinofugal pathways.

During ontogeny retinal ganglion cells manifest pronounced changes in excitable membrane properties. To further our understanding of the ionic conductances underlying such functional changes, the whole-cell voltage-clamp variation of the patch-clamp technique was used to record potassium currents in 220 ganglion cells dissociated from cat retinas ranging in age from embryonic day 31 to postnatal day 10. Potassium currents were isolated by blocking voltage-gated Na+ and Ca2+ currents with tetrodoxin (TTX) and CoCl2 respectively and were characterized by their pharmacology, kinetics and voltage-dependence of activation and inactivation. In all cases, a combination of three currents accounted for the total outward calcium-independent K+ current: (i) a steady linear conductance; (ii) a voltage-gated transient current, IA, and (iii) a voltage-gated sustained current, IK. Both voltage-gated currents were affected by the application of 4-aminopyridine and tetraethylammonia (TEA): IA showed a greater sensitivity to 4-aminopyridine, while IK was more sensitive to TEA. Both voltage-gated currents were present throughout the developmental period examined; however, the percentage of retinal ganglion cells (RGCs) expressing IA showed a marked decline from 82% at E31 to 45% at postnatal ages. During this developmental period there was an increase in the density of the two voltage-gated and the linear conductance. Additionally, with maturation, significantly slower inactivation kinetics were observed for IK. These findings, and our previous results dealing with maturational changes in the TTX-sensitive voltage-gated Na current, are related to the generation of excitability in developing retinal ganglion cells. Furthermore, the presence of cells with and without transient K+ conductance throughout development suggests that the different spiking patterns observed in RGC classes may be partially due to differences in their membrane properties.

Aging↗
PubMed1995

Distinct temporal patterns of expression of sodium channel-like immunoreactivity during the prenatal development of the monkey and cat retina.

Polyclonal and monoclonal antibodies prepared against the alpha-subunit of the voltage-gated sodium channel (alpha NaCh) were used to examine the distribution of sodium channel-like immunoreactivity during the prenatal development of the cat and rhesus monkey (Macaca mulatta) retina. At all prenatal ages studied, beginning on embryonic day 29 (E29) in the cat and E52 in the monkey, both antibodies labelled optic axons. With the polyclonal antibodies, the appearance of positive cells largely mirrored the onset of their morphological maturation. Immunoreactivity appeared first in the somata of ganglion cells, and subsequently the inner plexiform layer could be distinguished by its intense immunolabelling. A few weeks later horizontal cells displayed immunolabelling that extended to their dendrites in the developing outer plexiform layer. This was followed by immunoreactive cones, with bipolar cells labelled only postnatally. By contrast, with the monoclonal antibody some cells were found to be immunoreactive while their somata were still in the ventricular layer (E33 in cat and E52 in monkey). Many of these cells appeared to migrate to the outer portion of the prospective inner nuclear layer, where they gradually acquired the morphological appearance of bipolar cells. Transient expression of immunolabelling with monoclonal sodium channel antibody was found in the cones of the cat and cones and rods of the monkey. These results indicate that different types of alpha NaCh-like proteins are expressed in the mammalian retina at distinct developmental periods. Their presence at very early stages during development suggests that these proteins could play a specific role in the commitment and/or differentiation of specific retinal cell types.

Animals↗
PubMed1995

Development and regulation of dendritic stratification in retinal ganglion cells by glutamate-mediated afferent activity.

In the mature retina, the dendrites of retinal ganglion cells (RGCs) are segregated into either ON or OFF sublaminae of the inner plexiform layer (IPL), but early in development the dendritic processes of these cells are multistratified, ramifying throughout the IPL. We examined the time course of dendritic stratification in developing beta cells, the largest class of ganglion cells in the cat retina, by retrograde labeling of fixed tissue with Dil. Dendritic stratification begins in the central and peripheral retina by embryonic day 50, about 2 weeks before birth and is not fully completed until 5 months postnatally. A clear central-to-peripheral gradient in the incidence of stratified beta cells first becomes evident shortly after birth. This stratification process was effectively halted by short-term intraocular injections (4-11 d) of the glutamate analog 2-amino-4-phosphonobutyrate (APB), which hyperpolarizes rod bipolar cells and ON cone bipolar cells, thereby preventing the release of glutamate by these interneurons. APB treatment did not alter the somal sizes or the tangential extent of the dendrites of developing beta cells, nor did it cause abnormal loss of these neurons. The organization of the inner nuclear layer, containing the APB-sensitive bipolar cells, was also not compromised by such injections. When APB treatment was discontinued there was a rapid resumption of dendritic stratification resulting in a normal incidence of stratified RGCs. Thus, short-term APB treatment causes a delay rather than a permanent arrest of the stratification process.(ABSTRACT TRUNCATED AT 250 WORDS)

Afferent Pathways↗
PubMed1994

Neuropeptide Y immunoreactivity identifies a regularly arrayed group of amacrine cells within the cat retina.

Retinal amacrine cells can be divided into subgroups on the basis of morphological properties and chemical content. It is likely that these subgroups have specific connections and serve unique functional roles within the inner plexiform layer. In the present study we show that immunoreactivity to neuropeptide Y (NPY) identifies a group of amacrine cells (165,000-170,000) within the adult cat retina. This is the largest group of peptide-containing amacrine cells identified to date in the cat retina. These neurons have small cell bodies and are regularly spaced at all retinal eccentricities examined. The density of NPY-immunoreactive cells, as well as their regular spacing, suggests that these neurons form a specific subgroup of the amacrine cell class and are likely to serve a unique role in the transfer of visual information through the inner plexiform layer.

Animals↗
PubMed1994

Organization of pioneer retinal axons within the optic tract of the rhesus monkey.

Retinal ganglion cell axons must make a decision at the embryonic optic chiasm to grow into the appropriate optic tract. To gain insight into the cues that play a role in sorting out the crossed from the uncrossed optic axons, we investigated the sequence of their initial ingrowth in rhesus monkey embryos. Two carbocyanine dyes, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate and 4-(4-dihexadecylaminostyryl)-N-methylpyridinium iodide, were placed, respectively, into the left and right retinas to identify the course of uncrossed and crossed retinal axons through the optic chiasm and tract. Our results show that at embryonic day 36 the most advanced retinal projections are uncrossed. At this age the leading crossed axons are just reaching the chiasmatic midline, whereas the uncrossed fibers have already entered the optic tract. This indicates that the pathfinding of these pioneer uncrossed fibers does not require the presence of retinal axons from the opposite eye. At subsequent stages of development (embryonic days 40 and 42) there is a clear partial segregation of the uncrossed and crossed retinal axons within the optic tract: the uncrossed-component course is in the deeper portion of the optic tract, whereas the crossed component lies in a more superficial region. Thus, the spatial organization of retinal axons within the primordial optic tract reflects the sequential addition of the uncrossed and crossed retinal fibers. The orderly and sequential ingrowth of these pioneer retinal axons indicates that specific chiasmatic cues are expressed early in development and that such pioneer fibers may serve as guides for the later-arriving retinal fibers.

Animals↗
PubMed1993

Neuropeptide Y immunoreactivity identifies a group of gamma-type retinal ganglion cells in the cat.

Ganglion cells within the cat retina have been traditionally grouped by morphological criteria into three major classes: alpha, beta, and gamma. The gamma-type cells have been least well characterized, but the available evidence indicates that this class comprises a relatively heterogeneous population of neurons. In the present study we demonstrate that an antibody for neuropeptide Y (NPY) recognizes a subpopulation of about 2,000 gamma-type ganglion cells. The NPY-immunoreactive (IR) neurons project to the superior colliculus and to the C layers of the lateral geniculate nucleus as demonstrated by retrograde labeling with fluorescent tracers (fluorogold or rhodamine latex microspheres). Virtually all of these cells disappear following lesions of the optic nerve. The NPY-IR ganglion cells were identified as gamma cells on the basis of soma size and dendritic branching patterns. The somas of these neurons are small (8-22 microns in diameter), and each cell is characterized by sparsely branching dendritic processes, usually extending into the middle third of the inner plexiform layer, the physiologically defined ON sublamina. These neurons are distributed across the entire retina, with the highest density at the area centralis. Within local regions of the retina, however, there was no indication that the NPY-IR gamma cells are arrayed in a regular mosaic pattern. These results provide the first evidence that the gamma class of ganglion cells of the cat retina can be subdivided on the basis of immunocytochemical properties.

Animals↗
PubMed1993

Stratification of ON and OFF ganglion cell dendrites depends on glutamate-mediated afferent activity in the developing retina.

A fundamental attribute of the vertebrate visual system is the segregation of ON and OFF pathways signalling increments and decrements of light. In the mature retina, dendrites of ON- and OFF-centre retinal ganglion cells (RGCs) stratify in different sublaminae of the inner plexiform layer (IPL), and are differentially innervated by two types of bipolar cells which depolarize and hyperpolarize on exposure to light. This stratification of ON and OFF RGCs is achieved by the gradual restriction of their dendrites which ramify throughout the IPL early in development. The factors underlying this regressive event are unknown. Dendritic stratification occurs around the time that bipolar cells form synapses in the IPL, which raises the possibility that synaptic activity is involved in this process. Here we test this hypothesis by treating the developing cat retina with the glutamate analogue 2-amino-4-phosphonobutyric acid (APB), which hyperpolarizes ON cone bipolar and rod bipolar cells, thereby preventing their release of glutamate. We report that intraocular injection of APB during the period when dendritic stratification normally occurs prevents the formation of structurally segregated ON and OFF retinal pathways. These results provide evidence that glutamate-mediated afferent activity regulates the remodelling of RGC dendrites during development.

Aminobutyrates↗
PubMed1993

Prenatal development of excitability in cat retinal ganglion cells: action potentials and sodium currents.

The development of precise retinofugal projections is dependent on activity-mediated events, but as yet nothing is known about the ontogeny of excitable membrane properties in retinal ganglion cells (RGCs). In order to begin to understand how functional maturity is attained in these neurons, whole-cell patch-clamp recordings were obtained from acutely dissociated RGCs of fetal and postnatal timed-pregnant cats. Current-clamp recordings revealed a pronounced developmental increase in the proportion of RGCs capable of generating action potentials. At embryonic day 30 (E30), 5 weeks before birth and during a time when RGCs are still being generated, electrical stimulation elicited spikes in only a third of the cells. None of these neurons were capable of multiple discharges in response to maintained depolarization. The proportion of spiking neurons increased during ontogeny, such that by E55 all RGCs could be induced to generate action potentials, with the majority manifesting repetitive spiking patterns. Application of tetrodotoxin abolished spike activity of all fetal RGCs, indicating that sodium-mediated action potentials are present very early in development. At the same time, voltage-clamp recordings revealed significant ontogenetic modifications in several key properties of the sodium currents (INa). These were (1) a twofold increase in Na current densities; (2) a shift in the voltage dependence of both activation and steady state inactivation: with maturity, sodium currents activate at more negative potentials, while steady state inactivation of INa occurs at less negative potentials; and (3) a decrease in decay time constants of the Na current, at membrane potentials negative to -15 mV. These developmental changes were largely restricted to the period of axon ingrowth (E30-E38), suggesting that maturation of INa is not the limiting factor for the onset of activity-dependent restructuring of retinofugal projections.

Action Potentials↗
PubMed1992

Ontogeny of somatostatin immunoreactivity in the cat retina.

In the ganglion cell layer of the adult cat retina, subgroups of displaced amacrine cells and alpha ganglion cells are immunoreactive for somatostatin or a somatostatinlike substance. Both types of immunoreactive cells are found preferentially in inferior retina. We studied the development of somatostatin immunoreactivity in the prenatal and postnatal cat retina to determine how such unusual distributions of immunoreactive cells arise. Somatostatin-immunoreactive profiles were first observed at embryonic day (E) 30, within the inner retina in a central region that included the optic disk and the area centralis. By E36, immunoreactivity had virtually disappeared from the central retina but was present throughout the periphery. The immunoreactive profiles could not be classified morphologically because of their immaturity but were most likely retinal ganglion cells, the earliest born cells of the inner retina. Of the two types of immunoreactive cell observed in the adult, the first to be recognized morphologically was the displaced amacrine cell, at E45. These cells were virtually adultlike in morphology and number by E51, two weeks before birth. In contrast, immunoreactive alpha ganglion cells were not apparent until five days after birth and did not achieve their mature numbers and immunoreactive staining characteristics until more than a month later. From the time they could initially be recognized, both immunoreactive displaced amacrine cells and alpha cells were distributed mainly in the inferior retina. A third type of somatostatin-immunoreactive cell was transiently observed in the superior and inferior retina during prenatal and early postnatal development. These cells were characterized by granular staining in irregular shapes and few, if any, faintly stained processes. Injections of retrograde tracers into retinorecipient targets revealed that many of these cells were retinal ganglion cells. They disappeared by postnatal day 38. Our results indicate that somatostatin immunoreactivity initially follows a central-to-peripheral pattern of development, as is typical of other developmental events in the mammalian retina. They also indicate that the two types of somatostatin-immunoreactive neurons present in the adult cat retina (displaced amacrine and alpha ganglion cells) attain their mature immunocytochemical properties with very different timecourses. Finally, the observation that somatostatin immunoreactivity appears transiently in the granular-staining ganglion cells, distributed throughout the superior and inferior retina, suggests that the peptide may play a regulatory role in the development of the retina and/or retinofugal pathways.

Animals↗
PubMed1991

Substance P immunoreactivity identifies a projection from the cat's superior colliculus to the principal tectorecipient zone of the lateral posterior nucleus.

Cells in the superficial layers of the superior colliculus innervate multiple visual regions within the pulvinar-lateral posterior complex of the cat. To characterize these neurons we have examined their immunocytochemical properties in conjunction with their projection patterns. In the present study, we show that a monoclonal antibody for substance P recognizes a morphologically diverse population of neurons, which can be classified as granular, stellate, angular, and horizontal or nonhorizontal fusiform cell types. These neurons are distributed throughout the superficial layers of the colliculus, with a peak density corresponding to sublayer 2 of the stratum griseum superficiale. Injections of rhodamine latex micropheres into the pulvinar-lateral posterior complex demonstrate that a substantial proportion of these collicular cells (at least 35%) project to this region of the posterior thalamus. The overall population of substance P-containing cells, as well as the immunoreactive projection neurons, is composed of the same proportions of cell classes, with the exception that granular cells were not found to be projection neurons. A distinct wedge of substance P immunoreactivity, consisting of fiber and diffuse extracellular labeling, was discovered in the pulvinar-lateral posterior complex. This staining was demonstrated to be confined entirely within the medial division of the lateral posterior nucleus, which is considered to be the principal tectorecipient zone of the extrageniculate visual thalamus. Lesions of the superior colliculus largely abolished the substance P immunoreactivity in the ipsilateral tectorecipient zone. These results are consistent with the view that substance P plays a role in the functional organization of the principal tectothalamic pathway of the cat's extrageniculate visual system.

Animals↗
PubMed1991

Subgroup of alpha ganglion cells in the adult cat retina is immunoreactive for somatostatin.

We have previously shown that two types of cells in the ganglion cell layer of the adult cat retina are immunoreactive for somatostatin (White et al., '90). One of the types was identified by morphological criteria as a wide-field amacrine cell. The other cell type had a large, angular soma that resembled the alpha ganglion cell, but evidence was not available to identify it definitively as a ganglion cell. Both cell types were distributed preferentially in the inferior retina. In this report, we demonstrate that the two types of cell are, indeed, displaced amacrine cells and alpha ganglion cells. First, when retrograde tracers were injected into central visual targets, the immunoreactive large cells but not the displaced amacrine cells were found to be labeled. Second, after unilateral section of the optic nerve, the immunoreactive large cells disappeared from the retina on the lesioned side, but the displaced amacrine cells occurred in the same numbers in both retinae. In the periphery, the large cells ranged in diameter from 33 to 47 microns, comparable only to alpha ganglion cells (Boycott and Wässle, '74). An antiserum to parvalbumin was used to visualize the dendrites (Röhrenbeck and Wässle, '88) of somatostatin-immunoreactive large cells. Based on dendritic stratification within the inner plexiform layer (Famiglietti and Kolb, '76), the somatostatin-immunoreactive large cells were found to include both on-center cells and off-center cells, but were predominantly of the off-center type. Within a local region, they were found to be arrayed with greater regularity than the overall population of alpha ganglion cells. These results indicate that alpha ganglion cells of the cat retina can be subdivided on the basis of their immunoreactive staining for somatostatin and suggest that the diversity of ganglion cells in the cat retina may be greater than has been recognized on the basis of morphological criteria alone.

Animals↗
PubMed1991

The nasotemporal division of retinal ganglion cells with crossed and uncrossed projections in the fetal rhesus monkey.

The development of the partial decussation pattern in the primate retina was studied in fetal rhesus monkeys of known gestational ages. Retinal ganglion cells with either crossed or uncrossed projections were identified by labeling with HRP following unilateral injections of this tracer into the optic tract. At all fetal ages, very few cells (less than 0.5% of the total ganglion cell population) were found to project to the inappropriate hemisphere. The nasotemporal overlap zone, defined as the retinal region along the vertical meridian containing cells with either crossed or uncrossed projections, also appeared equivalent to that described for the adult animal. A temporal offset in the decussation pattern of large ganglion cells, similar to that of the mature retina, could be recognized as early as 50 d before birth. These results indicate that an adultlike retinal decussation pattern is evident in the fetal primate at a stage when projections from the 2 eyes are completely intermingled within retinorecipient nuclei, and prior to the onset of retinal ganglion cell loss. Moreover, the primate visual system exhibits a degree of precision in the specification of the nasotemporal division unrivaled among the mammalian species studied to date. The developmental specificity evident in the decussation pattern of the fetal rhesus monkey appears to reflect the specialized organization of this primate's retina for binocular focal vision.

Animals↗
PubMed1990

Somatostatin-immunoreactive cells in the adult cat retina.

Peptides have been found in the retinas of all mammalian species studied to date, but little is known about their localization and function in the cat. Using two mouse monoclonal antibodies directed to somatostatin 14, we have observed two sparse groups of somatostatin-immunoreactive neurons in the cat, both distributed preferentially in the inferior retina. The more numerous cell type is characterized by a small- to medium-sized soma (mean diameter = 16.3 +/- 9.0 microns; n = 186) with sparsely branching, far-reaching varicose processes that ramify mainly in the inner plexiform layer. The majority of these cells are located in the ganglion cell layer, with the remainder in the proximal inner nuclear layer and the inner plexiform layer. They are in especially high density at the retinal margin. In morphology and soma size, these cells resemble wide-field amacrine cells. The second cell type has a large, granular-staining soma (mean diameter = 29.7 +/- 14.8 microns; n = 145) with poorly stained primary processes and is found only in the ganglion cell layer. Cells of this type are most similar in their size and morphology to alpha ganglion cells. In contrast to the location of somatostatin-immunoreactive somata, a dense meshwork of immunoreactive processes was observed at all eccentricities within the inner plexiform layer, adjacent to the inner nuclear layer and to the ganglion cell layer. Labeled processes arising from the inner plexiform layer were also occasionally detected in the outer plexiform layer and the nerve fiber layer. Additional processes of unknown origin were observed in the nerve fiber layer and the optic nerve head. The extensive distribution of immunoreactive processes suggests that somatostatin-immunoreactive somata located preferentially in the inferior half of the retina have a widespread influence on neural activity.

Animals↗
PubMed1989

Callosal projection neurons in area 17 of the fetal rhesus monkey.

We have studied the distribution of callosal projection neurons in area 17 of a fetal rhesus monkey which received large injections of horseradish peroxidase into the contralateral occipital cortex. In comparison to other cortical areas, area 17 contains few callosal projection neurons. Most of these cells are confined to a region extending tangentially about 2.5 mm from the 17/18 border, although a few neurons were noted as much as 5 mm from the border. Comparing the distribution of callosal projection neurons in the fetal monkey with what has been described in newborn and adult macaques, it is apparent that although some degree of refinement in striate callosal connections may occur during in utero development, the prenatal development of callosal connections in the macaque is inherently adult-like.

Animals↗