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Biomedical subjects

A W Spira

Publications and source records attributed to A W Spira.

At least 19 recordsLinked to original sources

Computer managed instruction in a "clinical presentation-based" curriculum.

Many institutions are currently in the process of reorganizing their medical curricula from various traditional forms to a problem-based format. This method of instruction inevitably leads to a shift from traditional lecture and laboratory sessions to a more student oriented, small group session--a self-guided learning approach. This paper discusses the development and use of a system designed to provide the student with the necessary tools to access and control their learning environment.

Alberta↗

Effects of dark-rearing on the retinal dopaminergic system in the neonatal and postnatal guinea pig.

To appraise the extent of light responsivity of the retinal dopaminergic system in the early postnatal period, dopamine content (DA) of retinas of cycling-light-reared and dark-reared guinea pigs was determined. DA storage postnatally was suppressed in dark-reared animals. Their retinas responded to a brief light stimulus by rapidly increasing DOPA synthesis and DA utilization, but not to the extent of the response of cycling-light-reared animals. Dopaminergic retinal neurones depend on light stimulus for the complete development of some of their neurochemical characteristics.

Animals↗

Cellular localization of somatostatin mRNA in rat retina.

In an attempt to determine the localization of the messenger RNA (mRNA) encoding somatostatin in the rat retina, we studied Sprague-Dawley rats by in situ hybridization histochemistry using radiolabelled oligodeoxyribonucleotides complementary for rat somatostatin mRNA. Among the layers of retina, we found specific labelling in the soma of some cells in the innermost and outermost laminae of the inner nuclear layer and in the ganglion cell layer; no specific labelling was observed in the inner and outer plexiform layers or in the outer nuclear layer. These data indicate the major site of somatostain synthesis within the rat retina.

Animals↗

Ontogenesis of the electroretinogram in a precocial mammal, the guinea pig (Cavia porcellus).

1. Ontogenesis of the electroretinogram, the mass electrical response of the retina to flash light stimuli, was studied in the guinea pig (Cavia porcellus), a precocial species with visual function at birth. 2. a-Wave components, b-wave, oscillatory potentials, slow PIII, and c-wave responses to flash stimuli developed between 55 and 64 days of gestation (full term is 68-69 days). 3. a-Waves attributable to photoreceptor functions were fully mature at 60 days. 4. ERG development lagged behind the reported critical milestones in retinal development; its prenatal onset indicates that no history of light entrainment is required for initiation of a mature ERG response.

Aging↗

Histological and electron microscopic milestones in the development of the retina of a marsupial wallaby, Macropus eugenii.

Retinal differentiation in the pouch young of the wallaby Macropus eugenii was characterized microscopically and morphometrically. Mitosis occurs until early in the second month in the central retina, and until early in the fourth month, peripherally. Separation of the neuroblast layer by the outer plexiform layer did not immediately halt cell division. The retinal surface continued to expand well past the time of cessation of proliferation. Cell death in the ganglion cell layer continued through the fourth month centrally and to nearly five months in the periphery. The major period of cell death was coincident with the segregation of retinal afferents and the refinement of topography in the superior colliculus and dorsal lateral geniculate nucleus. Beginning in the third month retinal thickness, measured between the outer limiting membrane and nerve fiber layer, declined equally in peripheral and central regions. At all stages the combined thicknesses of the outer and inner nuclear layer in the retinal periphery was greater than that in the center. Together with a late thickening of the inner plexiform layer, the data are consistent with the suggestion that expansion of peripheral non-ganglion cell elements may play a role in development of center to periphery differences in ganglion cell distributions. Retinal differentiation of the wallaby follows the pattern of most mammals. The onset of development of key milestones for the acquisition of retinal function occurred in the sequence: conventional synapse formation prior to ribbon synapse formation in the inner plexiform layer, and photoreceptor outer segment differentiation prior to terminal triad synapse formation.

Animals↗

The effects of two anticonvulsants on amino acid levels in the developing rat cerebellum.

Two anticonvulsants were administered pre- and postnatally to determine their effects on putative amino acid neurotransmitter levels in the rat cerebellum. The amino acids were quantitated using precolumn fluorescence derivatization and reverse-phase high performance liquid chromatography at various postnatal intervals. Treatment with clonazepam produced an initial depression in levels of most of the amino acids analyised. By three weeks postnatal all the amino acids, with the exception of GABA, had returned to control levels. GABA levels were still depressed five weeks after the cessation of treatment. Phenobarbital treatment produced an initial elevation in the level of GABA. At three weeks postnatal, both GABA and glutamate levels were elevated and remained so at eight weeks postnatal. In conclusion, the data demonstrated that each anticonvulsant produced unique, acute and chronic alterations in the levels of the cerebellar amino acids.

Amino Acids↗

Effect of phenobarbital on Purkinje cell growth patterns in the rat cerebellum.

The effects of low level phenobarbital administration (18 postcoitus to 21 days postnatal) on Purkinje cell growth and remodeling were studied from 3 to 20 weeks postnatal). The Purkinje cell dendritic trees were analyzed both metrically and topologically using the method of vertex analysis. The total segment length, mean terminal path length, and mean vertex path length were reduced in the treated cells. The pattern of segment frequency as related to equivalent orders was abnormal in the treated cells. The Va/Vb vertex ratios and the levels of trichotomy indicated that the treated cells underwent nonrandom remodeling, unlike the control cells which exhibited dichotomous, random terminal branching. These observations confirm that phenobarbital produces distinct long-term morphologic alterations in Purkinje cells.

Animals↗

The effect of chlorpromazine on monoamine levels in the rat cerebellum and hippocampus: a developmental study.

Acute and chronic alterations of the monoamine levels in the rat cerebellum and hippocampus after late prenatal and early postnatal administration of chlorpromazine, were examined using high performance liquid chromatography with electrochemical detection. During a critical phase of development (one week postnatal), chlorpromazine produced increased levels of both serotonin and dopamine in both brain regions. At 24 weeks postnatal, or 21 weeks after cessation of drug administration, the levels of all 3 monoamines were decreased in the cerebellum and increased in the hippocampus. The early disruption in the normal monoamine levels may be related to abnormal growth patterns observed in previous studies.

Animals↗

Localization of choline acetyltransferase-like immunoreactivity in the embryonic chick retina.

Putative sites of acetylcholine synthesis in the retina of the embryonic and posthatched chick were localized immunohistochemically with antisera to choline acetyltransferase; the resultant choline acetyltransferase-like immunoreactivity (ChAT-IR) was compared to demonstrated sites of acetyltransferase (AChE) activity, and changes were followed in localization during development. The results confirmed the early and rapid course of development of the chick's retinal cholinergic system described in previous biochemical and morphological studies. Immunoreactivity was first detected at embryonic day 6.5 in cells close to the retina's vitreal surface. By 8 days it was present in cells in two juxtaposed rows; by the ninth day the two rows were separated and immunoreactivity was evident in two subliminae of the inner plexiform layer. On the tenth day distribution was like that in the posthatched chicken, in type I cholinergic cells in the inner nuclear layer and in type II cells in the ganglion cell layer (Millar et al.: Neurosci. Lett. 61:311-316, '85), and similar to that of most vertebrates. Three days before hatching, a third population of weakly immunoreactive cells (type III cells) appeared within the inner nuclear layer. The onset of localizable ChAT-IR occurred in amacrine cells and in their processes, before the period of synaptogenesis. Acetylcholinesterase activity was localized at an earlier age than ChAT-IR, and at all ages was present in more cells. The results obtained support the view that "displaced" cholinergic amacrine cells begin to differentiate at the same time and in the same retinal region as type I cholinergic cells. Separation of the two groups is a consequence of the ramification of processes of amacrine and ganglion cells rather than a result of the secondary migration of cells between layers.

Acetylcholinesterase↗

Effect of chlorpromazine on Purkinje cell growth patterns.

The phenothiazine, chlorpromazine, when administered perinatally to rats is implicated in the production of both motor and behavioural deficits. In this study the effects of chlorpromazine on dendritic growth of cerebellar Purkinje cells were examined after administering the drug to litters of Long-Evans hooded rats from 18 days postcoitus to 21 days postnatum. Various parameters of Purkinje cell growth were analysed at specific times after treatment utilizing Golgi-prepared specimens. Granule cell density was used as an estimate of parallel fibre numbers. When compared with control Purkinje cells the treated cells at 7 weeks postnatum had established longer segments and increased mean vertex path length. The total number of segments was also increased and the distribution of segments of different orders was abnormal. In addition, the spines on terminal dendritic branches were both longer and of increased density in the chlorpromazine-treated group. These observations confirm that chlorpromazine produces distinct morphological alterations in mature Purkinje cells.

Animals↗

Localization, chromatographic characterization, and development of somatostatin-like immunoreactivity in the guinea pig retina.

We have studied the anatomical localization, chromatographic properties, and development of somatostatin-like immunoreactivity (SLI) in the guinea pig retina. The majority of guinea pig retinal SLI in the adult and fetus eluted similarly to somatostatin-28 by gel filtration and high pressure liquid chromatography. This represents a higher ratio of somatostatin-28-like material to total SLI than had been observed in retinal extracts from most other animal species. Somatostatin-like-immunoreactive cells and fibers were localized using two antisera in cryostat-sectioned and flat mounted retinae. Distribution of cells and fibers differed uniquely from that in other species previously reported. Reactive perikarya were located only in the far peripheral region: in the innermost layer of the inner nuclear layer (INL), in the inner plexiform layer (IPL), and in the ganglion cell layer. Reactive fibers were prominent in the IPL and nerve fiber layer (NFL) in both the peripheral and central retina. Less frequently, processes were observed between the NFL and IPL, between the IPL and INL, and, rarely, in the outer plexiform layer and outer nuclear layer. Small numbers of reactive fibers were found in the optic nerve and disc. These observations suggest that processes of intrinsic (amacrine or associational ganglion cells) and projection neurons (true ganglion cells or efferent fibers) containing SLI are intermingled in the guinea pig retina. SLI, quantified by radioimmunoassay, was present in the developing retina as early as the 6th week of gestation (full term is 10 weeks). Immunohistochemically detected somatostatin-like-immunoreactive elements were seen first at 2 weeks before birth, coincident with the onset of the period of most rapid increase in levels of assayed SLI. Somatostatin-like-immunoreactive peptides reached two-thirds of adult levels by birth.

Animals↗

Effects of alteration in the light cycle on outer segment shedding in the fetal retina.

Shedding of outer segment discs and their phagocytosis by retinal pigment epithelial cells (RPE) in the guinea pig starts to occur during in utero development. The number of phagosomes in the fetal RPE varies in a cyclical manner somewhat analogous to that observed in mature animals. The present study addresses the question of whether the cycles in fetus and mother are independent or linked. Pregnant guinea pigs were subjected to 10 days of alteration of their lighting schedules beginning on the 46th day of gestation. Dams and fetuses were sacrificed on day 56, and large phagosomes in the RPE of each were counted. A 10-hour phase advance was employed, ie, from 0800 (lights on): 2000 (light off) in controls, to 2200 (on): 1000 (off) in experimental animals. Counts of large phagosomes in the posterior retina were compared in both control and experimental nulliparous, gravid, and fetal animals. A significant burst of shedding occurred within one hour after light onset in the experimental groups comprised of either nulliparous or pregnant animals. The latter also showed a smaller rise in phagosome numbers at the original time of onset of illumination. In contrast, no shift in the time of maximum shedding activity was observed in fetuses in the experimental regime. Their activity was similar to that present in the original 0800:2000 cycle. Whereas maternal RPE shedding cycles can be reentrained in a 10-day period, those in the 46- to 56-day fetal retina are refractory to significant change. Based on this finding, it is likely that the shedding cycles in the mother and fetus at this age are under independent control, and, therefore, that shedding in the fetus is not mediated by maternal borne factors.

Animals↗

The effects of chlorpromazine and phenobarbital on cerebellar Purkinje cells.

The morphological effects of two chemically different neuroactive drugs (chlorpromazine and phenobarbital) on developing Purkinje cells in the rat cerebellum were examined to determine the presence of cytological alterations. Therapeutic dosages of both drugs were chronically administered to separate groups of maternal rats beginning on day 18 postcoitus. Entire litters were sacrificed on postnatal days 13, 15, 18, and 21. Light microscopic quantitation of Purkinje cells demonstrated a statistically significant reduction in total numbers below control levels at all ages examined for both drugs. Pyknotic Purkinje cells, which appeared more numerous in the drug tested groups, had a paucity of synaptic contacts on both the soma and dendritic branches. Both drugs tested produced similar results within the parameters examined. These alterations have been discussed in relation to possible mechanisms and sites of action.

Animals↗

The effects of chlorpromazine and phenobarbital on vasculogenesis in the cerebellar cortex.

The morphological effects of two chemically different neuroactive drugs (chlorpromazine and phenobarbital) on vasculogenesis in rat cerebellum were examined to determine the presence of vascular alterations. Therapeutic dosages of both drugs were chronically administered to separate groups of maternal rats beginning on days 10, 13, 15, 18, and 21. In chlorpromazine-treated animals the specific length of blood vessels was most severely reduced in the Purkinje cell layer. Animals treated with phenobarbital demonstrated an initial reduction in specific length in the Purkinje cell layer but returned to control values by day 21 postnatal (p.n.). Blood vessels in the molecular and granular layers showed little change. The observed changes have been discussed in relation to possible mechanisms and their relationship to neurogenesis.

Animals↗

Phagocytosis in the fetal pigment epithelium: evidence for cyclic activity.

Photoreceptor cells in guinea pig retinas begin to shed their outer segments while developing in utero. To ascertain whether phagocytosis of outer segments in the fetus is a cyclical event like that of mature animals, large phagosomes in fetal retinal pigment epithelial cells were counted at representative times during a 24 hr period. These data were then compared with similar data obtained from the pregnant dams. Gravid animals and fetuses both showed a burst of shedding after the onset of illumination. Shedding was well initiated in the pregnant animals 30 min after the onset of illumination but was delayed for 2 to 3 hr in their fetuses. Thereafter the two patterns of phagocytosis showed substantial differences. Congruent cyclical patterns could be discerned, however, by graphing fetal counts with an advance of 10 hr on the time scale relative to that of the gravid animals. The results presented here represent the first indication of a cyclical pattern of phagocytosis in a population of fetuses during retinal development.

Animals↗

An ethanolic phosphotungstic acid (EPTA) analysis of photoreceptor and synaptic ultrastructure in the guinea pig retina.

Ethanolic phosphotungstic acid (EPTA) has been used to elucidate the structure of certain organelles contained within retinal cells not clearly discernible using conventional preparations. Both synaptic and nonsynaptic components of the guinea pig neural retina have been analyzed. Within the photoreceptor (PR) cell EPTA-stained components include the connecting cilia, their basal bodies, and the root filament system. Cross-striated fibrillar organelles, similar in appearance to the root filaments, are also observed in the nuclear region, the synaptic terminal and other parts of the PR cell. The possible structural continuity and significance of these structures are discussed. Within retinal synapses of both the inner and outer plexiform layers, ribbons and associated paramembranous specializations are stained. The photoreceptor ribbons have a trialaminar structure with filamentous, tufted borders. Synaptic cleft material and postsynaptic densities are also stained. Bipolar cell synapses in the inner plexiform layer contain stained short ribbons as well as closely associated peg-like densities extending towards the presynaptic membrane.

Animals↗

The structure and distribution of the cross-striated fibril and associated membranes in guinea pig photoreceptors.

Examination of longitudinally and transversely sectioned photoreceptor cells of the guinea pig retina revealed an aggregate of thin filaments forming a single cross-striated fibril coursing through the full length of the non-receptor portion of the cell. The fibril begins as the ciliary rootlet from the region of the basal body of the connecting cilium. From the basal body it passes between the mitochondria of the ellipsoid and along the Golgi zone of the myoid region of the inner segment, narrowing from an irregularly shaped bundle to a ribbon-shaped aggregate. The fibril separates into discrete strands, each curving along the nucleus, reuniting into a single bundle to pass down the cell's axon, and terminating deep within the synaptic terminal. The fibril is flanked by two separate membranous saccules, each continuous along nearly its full length. The fibril's extensive course in the guinea pig and its association with continuous membranes necessitates a reexamination of earlier proposals for the function of cross-striated filamentous structures in photoreceptor cells.

Animals↗