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P Witkovsky

Publications and source records attributed to P Witkovsky.

At least 73 records · Page 4Linked to original sources
PubMed1981

Synapse formation and modification between distal retinal neurons in larval and juvenile Xenopus.

A serial section analysis of photoreceptor synaptic bases was undertaken in the clawed frog Xenopus laevis. The developmental period from tadpole stage 48 through metamorphosis was studied. Horizontal cells contacted rod and cone photoreceptors at ribbon synapses; the number of such contacts per receptor base was constant for rods, but increased for cones as a function of developmental stage. In pre-metamorphic animals bipolar cells contacted receptors only through basal junctions; their number in cone bases increased dramatically during development but was unchanged in rod bases. A densitometric estimation of the cleft width of basal junctions showed that it ranged from 10 to 18 nm, but the junctions could not be divided reliably into the 'wide' and 'narrow' categories reported for other vertebrate species. Near metamorphic climax a new type of ribbon-related bipolar cell junction appeared. Gap junctions between horizontal cells and conventional synapses of horizontal cell onto bipolar cell processes were first seen in mid-larval developmental stages.

Animals↗
PubMed1981

A freeze-fracture study of synaptogenesis in the distal retina of larval Xenopus.

Synapse formation between photoreceptor, bipolar and horizontal cells of the larval Xenopus retina was studied by the freeze-fracture technique. Photoreceptors and horizontal cells were joined by ribbon synapses; photoreceptor and bipolar cells by basal junctions. Gap junctions were found between photoreceptors and between horizontal cells. Horizontal cell dendrites invaginated receptor bases before the plasma membrane of either cell showed zones of intramembrane (IMP) particle accumulation. Subsequently the receptor cell began to form a synaptic ridge where P-face IMPs aggregated at a protrusion of the surface membrane. The length of the ridge and the density of its IMPs increased between larval stages 40 and 56. Cross-fractured views of receptor cytoplasm at different larval stages showed that synaptic ribbons and synaptic vesicles developed in conjunction with the ridge. Plasmalemmal deformations suggesting sites of vesicle fusion or uptake were noted adjacent to the apex of the ridge. Horizontal cell dendritic membrane first accumulated P-face IMPs at several small regions; subsequently the IMPs became aligned over a broad membrane area. Both rod- and cone-related horizontal cell dendrites also manifested a loose patch of E-face IMPs which subsequently was transformed into a linear array. Basal junctions were characterized by a P-face IMP aggregate in the photoreceptor membrane and an E-face IMP aggregate in the bipolar cell membrane. Basal junctions appeared suddenly in a mature configuration at larval stage 42.

Animals↗
PubMed1979

Synaptic connections linking cones and horizontal cells in the retina of the pikeperch (Stizostedion vitreum).

Cones and horizontal cells of the pikeperch retina were studied with morphological and physiological techniques. Gap junctions were observed between cone pedicles and basal processes emitted by neighboring cones. Intracellular recordings showed that the light-evoked hyperpolarizing cone response was enhanced by light falling upon neighboring receptors within a radius of 50 microns. We suggest that the network of gap junctions between cones mediates the summative lateral interaction described. Three sub-classes of horizontal cells (H1, H2, H3) send dendrites to cones; H1 and H2 cells appear to contact twin cones, exclusively or preferentially, whereas H3 cells appear to synapse only with single cones. Horizontal cells of the same sub-class are joined by gap junctions between dendrites or at the lateral faces of perikarya. These unions extend over several micron 2 and as seen in transmission electron microscopy consist of patches of close apposition alternating with areas of membrane separation, folding and occasional zonulae adherents. Freeze-fracture profiles of horizontal gap junctions show localized areas of dense particle aggregation on the P-face and pits on the E-face flanked by regions of unspecialized membrane. These morphological findings provide support for the known spatial and color-coding properties of pikeperch horizontal cells.

Animals↗
PubMed1978

The formation of photoreceptor synapses in the retina of larval Xenopus.

The first appearance and early development of synapses between photoreceptors, bipolar cells and horizontal cells were studied in the retina of larval Xenopus, between stages 37/8 and 46. Synapse morphology was reconstructed from examination of serial sections. Additional data were obtained from E-PTA stained tissue, Photoreceptors initially contacted horizontal cell dendrites via ribbon synapses at the photoreceptor basal surface (stages 37/8--39). Later, horizontal cell dendrites penetrated the receptor as digitiform processes (stages 39--40) which subsequently expanded extensively within the invagination (stages 40--44). At stages 41--46, a photoreceptor ribbon generally was associated with two horizontal processes in a synaptic dyad complex; each ribbon participated in 1 or 2 such complexes. Bipolar cell dendrites made contact with receptors via superficial or invaginating basal-type junctions. Such junctions were observed first at stage 39, but increased greatly in number beginning at stage 42. Basal junctions were characterized by marked staining of the paramembranous receptor cell cytoplasm and a 9--13 nm wide cleft containing an electron-dense material. No association of bipolar cell dendrites with photoreceptor ribbons was noted. Junctional complexes presumed to be gap junctions first were observed at stage 39 between photoreceptor bases and basal processes emitted by neighbouring photoreceptors.

Animals↗
PubMed1978

Rod sensitivity and visual pigment concentration in Xenopus.

Xenopus larvae were raised on a vitamin A-free diet under constant illumination until their visual pigment content had decreased to between 8% of normal and an undetectably low level. After the intramuscular injection of 2.1 X 10(13-2.1 X 10(16) molecules of [3H]vitamin A, ocular tissue showed a rapid rate of uptake of label which reached a maximum level of incorporation by 48 h. Light-microscopic autoradiography revealed that the retinal uptake of label was concentrated within the receptor outer segments. Spectral transmissivity measurements at various times after injection were made upon intact retinas and upon digitonin extracts. They showed that visual pigment with a lambdamax of 504 nm was formed in the retina and that the amount formed was a function of incubation time and the magnitude of the dose administered. Electrophysiological measures of photoreceptor light responses were obtained from the PIII component of the electroretinogram, isolated with aspartate. The quantal flux required to elicit a criterion response was determined and related to the fraction of visual pigment present. The results showed that rod sensitivity varied linearly with the probability of quantal absorption.

Animals↗
PubMed1978

Formation, conversion, and utilization of isorhodopsin, rhodopsin, and porphyropsin by rod photoreceptors in the Xenopus retina.

The visual pigment content of rod photoreceptors in Xenopus larvae was reduced greater than 90% through a combination of vitamin A-deficient diet and constant light. Thereafter, a dose of either all-trans-retinol or 9-cis-retinal was injected intramuscularly, leading to the formation of a rhodopsin (lambdamax 504 nm) or isorhodopsin (lambdamax 487-493 nm) pigment, respectively. Electrophysiological measurements were made of the threshold and spectral sensitivity of the aspartate-isolated PIII (photoreceptoral) component of the electroretinogram. These measures established that either rhodopsin or isorhodopsin subserved visual transduction with the same efficiency as the 519 nm porphyropsin pigment encountered normally. When animals with rhodopsin or isorhodopsin were kept in darkness or placed on a cyclical lighting regimen for 8 days, retinal densitometry showed that either pigment was being converted to porphyropsin; significantly more porphyropsin was formed as a result of cyclical lighting than after complete darkness.

Animals↗
PubMed1976

Electron microscopic observations of the mesencephalic nucleus of the fifth nerve in the Selachian brain.

The mesencephalic nucleus of the trigeminal nerve (mes V) in the brain of the skate (Raja oscellata) was studied by electron microscopy. Mes V neurons are large (40-80 mum diameter) and are located in the periventricular grey matter. Their perikaryal cytoplasm is rich in Golgi apparatus, small mitochondria, rough endoplasmic reticulum, polysomes and bundles of neurofilaments. A striking feature is the presence of masses of glycogen granules, at times surrounded by membrane wrappings and lysosomal bodies. Two types of conventional synaptic contacts were made onto mes V perikarya and dendrites. One had round, agranular vesicles and usually also contained dense-cored vesicles, the other had flattened, pleomorphic, agranular vesicles and usually lacked dense-cored vesicles. Additional membrane complexes consisting of a region of gap junction flanked by sites of desmosomal attachment were observed to link neighbouring mes V neurons. Somato-somatic, dendro-somatic, axo-somatic, and dendro-dendritic junctions were noted. Except for the somato-somatic union, one or more chemical synapses were located close to the sites of gap junctions.

Animals↗
PubMed1976

Photoreceptor thresholds and visual pigment levels in normal and vitamin A-deprived Xenopus tadpoles.

1. Measurements of the aspartate-isolated PIII component of the electroretinogram (ERG) were used to estimate photoreceptor threshold in dark-adapted tadpoles of the clawed toad, Xenopus laevis raised on a normal diet. Spectral sensitivity functions established that the rod mechanism governed absolute dark thresholds from stage 40 to the end of premetamorphic development. 2. Parallel measures of rod outer-segment dimensions and visual pigment levels demonstrated a) that visual pigment concentration remained constant at all tadpole stages, and b) that the fall in threshold over the course of premetamorphic development depended exclusively on the increased probability of quantal absorption that accompanied the growth of the rod outer segments. 3. When tadpoles were obtained from vitamin A-deficient females and raised subsequently on a vitamin A-free diet, the first appearance of the ERG was delayed and its absolute threshold raised, with respect to controls. 4. Histological examination of vitamin A-deprived retinas indicated that rod outer segments retained their structural integrity in spite of up to a 75% loss of visual pigment. 5. The threshold rise which accompanied a fall in visual pigment levels, whether effected by dietary deficiency or by bleaching, was greater than could be attributed solely to a reduction in the probability of quantal absorption.

Age Factors↗
PubMed1975

Slow PIII component of the carp electroretinogram.

The slow PIII component of the electroretinogram (ERG) was studied in the isolated, aspartate-treated carp retina. Although the latter is richly populated with cones, slow PIII appeared to reflect almost exclusively the activity of rods; e.g. the spectral sensitivity of the potential paralleled closely the rod pigment curve, its operating range (i.e. the V-log I curve) was limited to 3 log units above absolute threshold, and raising background intensities to photopic levels produced saturation of the increment threshold function without evidence of a cone-mediated segment. Only after bleaching away a significant fraction of the porphyropsin was it possible to unmask a small photopic contribution to slow PIII, as evidenced by a displacement in the action spectrum to longer wavelengths. The spatial distribution of the slow PIII voltage within the retina (Faber, D.S. 1969. Ph.D. Thesis. State University of New York. Buffalo, N.Y.; Witkovsky, P.J. Nelson, and H. Ripps. 1973. J. Gen Physiol. 61:401) and its ability to survive aspartate treatment indicate that this potential arises in the Müller (glial) fiber. Additional support for this conclusion is provided by the slow rise time (several seconds) and long temporal integration (up to 40s) of the response. In many respects the properties of slow PIII resemble those of the c-wave, a pigment epithelial response also subserved by rod activity. On the other hand, the receptoral (fast PIII) and the b-wave components of the ERG behave quite differently. Unlike slow PIII, response saturation could not be induced, since both potentials are subserved by cones when the stimulus conditions exceed the limits of the scotopic range. Receptors appear to govern light adaptation at photopic background levels; both fast PIII and b-wave manifest identical incremental threshold values over this range of intensities. However, under scotopic conditions, the sensitivity of the b-wave is affected by luminous backgrounds too weak to alter fast PIII threshold, indicating a postreceptoral stage of adaptation.

Adaptation, Ocular↗