Interreceptoral junctions in the teleost retina.
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Biomedical subjects
Publications and source records attributed to P Witkovsky.
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The mass photoreceptor response of the isolated carp retina was studied after immersing the tissue in aspartate-Ringer solution. Two electro-retinogram components were isolated by differential depth recording: a fast cornea-negative wave, arising in the receptor layer, and a slow, cornea-negative wave arising at some level proximal to the photoreceptors. Only the fast component was investigated further. In complete dark adaptation, its action spectrum peaked near 540 nm and indicated input from both porphyropsin-containing rods (lambda(max) approximately 525 nm) and cones with longer wavelength sensitivity. Under photopic conditions a broad action spectrum, lambda(max) approximately 580 nm was seen. In the presence of chromatic backgrounds, the photopic curve could be fractionated into three components whose action spectra agreed reasonably well with the spectral characteristics of blue, green, and red cone pigments of the goldfish. In parallel studies, the carp rod pigment was studied in situ by transmission densitometry. The reduction in optical density after a full bleach averaged 0.28 at its lambda(max) 525 nm. In the isolated retina no regeneration of rod pigment occurred within 2 h after bleaching. The bleaching power of background fields used in adaptation experiments was determined directly. Both rods and cones generated increment threshold functions with slopes of +1 on log-log coordinates over a 3-4 log range of background intensities. Background fields which bleached less than 0.5% rod pigment nevertheless diminished photoreceptor sensitivity. The degree and rate of recovery of receptor sensitivity after exposure to a background field was a function of the total flux (I x t) of the field. Rod saturation, i.e. the abolition of rod voltages, occurred after approximately 12% of rod pigment was bleached. In light-adapted retinas bathed in normal Ringer solution, a small test flash elicited a larger response in the presence of an annular background field than when it fell upon a dark retina. The enhancement was not observed in aspartate-treated retinas.
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1. Ganglion cell discharges were evoked by extrinsic polarization of the horizontal cells in the retina of the smooth dogfish (Mustelus canis). Depolarization of the horizontal cell gave rise to a discharge similar to that evoked by a spot of light (centre type response) and hyperpolarization of the horizontal cell, a discharge similar to that by an annulus (surround type response).2. Procion dye injection established that the current-passing electrode was sometimes located in the external horizontal cell. Other possibilities, such as middle and internal horizontal cells, were neither confirmed nor excluded.3. Activation of ganglion cells by current was possible under completely dark-adapted conditions and for several log units above this level.4. Depolarizing current enhanced the ganglion cell response evoked by a light spot in the centre of its receptive field; hyperpolarizing current antagonized the response to the same flash.5. The results are consistent with the supposition that a potential change in the horizontal cell, irrespective of its polarity, or whether produced by light or current, spreads within a laminar layer (the S-space). The effect of the potential change is to modulate the response of bipolar cells and their input into the ganglion cell.
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The aim of the present study was to determine the number of cone classes in the Xenopus retina. We examined the dimensions and staining properties of cones, utilizing two monoclonal antibodies, COS-1 and OS-2, developed by Szel and Rohlich (1985). Living cones also were reacted with the plant lectins peanut agglutinin (PNA) and wheat germ agglutinin (WGA) and with a fluorescent stilbene dye, DIDS, which binds selectively to red-sensitive cones (Kleinschmidt, 1991; Kleinschmidt & Harosi, 1992a,b). Three cone populations were distinguished based on differences in size and staining properties. Eighty-eight percent of all cones were stained strongly by COS-1, PNA, and DIDS, but weakly by OS-2. The group of cones stained by COS-1 had the largest mean dimensions of outer segment length, width, and oil droplet diameter. COS-1 negative cones were divisible into two groups: a subclass of miniature cones (approximately 4% total cones) was stained strongly by OS-2, PNA, and DIDS. The balance, constituting approximately 9% total cones, were of intermediate size, were not stained by PNA and reacted weakly to OS-2 and DIDS. WGA stained all cones. Large, COS-1+ cones appear to be red-sensitive and belong to the class of anion-tunable cone pigments. We suggest that the intermediate size, COS-1 negative cones are blue-sensitive based on the finding that blue-sensitive chromatic horizontal cells connect to them preferentially (Witkovsky et al., work in progress).(ABSTRACT TRUNCATED AT 250 WORDS)
Two types of spiking neuron were identified among acutely dissociated neurons from the Xenopus retina by their responses to a depolarizing current step: single spikers and multiple spikers. In culture, multiple spikers had perikaryal diameters > 15 microns, whereas single spikers had smaller somata, 5-10 microns in diameter. Using a conventional whole-cell patch-clamp technique, both T- and L-type calcium currents were identified in multiply spiking cells whereas only an L-type current was present in singly spiking cells. The metabotropic glutamate receptor (mGluR) agonist trans-(1S-3R)-1-amino-1,3-cyclopentane-dicarboxylic acid (trans-ACPD) significantly decreased the L-type calcium current by 46 +/- 3% (mean +/- S.E.M.) in both types of cell but had only a minor effect on the T-type current in multiply spiking neurons. In the presence of 50 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), 100 microM quisqualate (a potent mGluR1/5 agonist) decreased the L-type calcium current by 47 +/- 9% but had no effect on the T-type current. The selective mGluR4/6/7 agonist (+/-) 2-amino-4-phosphonobutyric acid (L-AP4, 100 microM), and the mGluR2/3 agonist (2S,3S,4S)-alpha-(carboxycyclopropyl)glycine (L-CCG1, 100 microM) decreased the L-type calcium current by 12 +/- 3% and 14 +/- 2%, respectively. The inhibition of calcium current by trans-ACPD was reduced when the patch pipette contained the G-protein inhibitor, GDP beta S. The presence of the G-protein activator GTP gamma S in the patch pipette irreversibly reduced the L-type calcium current, but was without effect on the T-type current. Heparin applied intracellularly significantly reduced the inhibitory effect of quisqualate, indicating an involvement of the inositol triphosphate (IP3) pathway in the mGluR-induced reduction of calcium current. Replacement of internal EGTA with BAPTA significantly reduced the inhibitory effect of quisqualate. In contrast, internal application of cAMP did not prevent an inhibition of calcium current by quisqualate. Thus, the mechanism by which calcium current is inhibited by mGluR seems not to involve an intracellular cAMP cascade. Our findings indicate that activation of mGluR1/5 results in the inhibition of a high-threshold calcium current. This process is mediated by the activation of a G-protein and is consistent with inhibition occurring by an IP3-stimulated release of internal calcium.