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Localization of immunoreactive cholecystokinin precursor to amacrine cells and bipolar cells of the macaque monkey retina.

We used antisera that recognized precursors of the neuropeptide cholecystokinin extended at the carboxyl terminus in an immunocytochemical study of the macaque retina. A subpopulation of bipolar cells with long, obliquely oriented dendrites was labeled. Their axons terminated exclusively in the fifth stratum of the inner plexiform layer, where they contacted processes of amacrine and ganglion cells. Based on their morphology, these cells appeared to be the type that contacts short-wavelength cones selectively. Two types of amacrine cells were also labeled, and processes from both types formed dense plexuses in the second and fourth strata of the inner plexiform layer. The majority of their synaptic connections were with other amacrine cells, but they had more contacts with bipolar cell axons and retinal ganglion cell dendrites than any other peptidergic cells in the macaque retina. We studied extracts of macaque retina with gel-filtration chromatography and radioimmunoassays to confirm our immunohistochemical results. We found cholecystokinin octapeptide and other immunoreactive forms that were amidated at their carboxyl termini and were therefore likely to be biologically active. Unlike most other regions of the CNS, however, the retina had relatively low concentrations of amidated forms, and forms with extended carboxyl termini that are presumably their precursors were far more abundant. These findings suggest that the rate of cholecystokinin synthesis in the retina is quite high, as we would expect if the peptide were found in tonically active neurons.

Amino Acid Sequence↗

The population of bipolar cells in the rabbit retina.

The population of bipolar cells in the rabbit retina was studied using Golgi impregnation and photocatalyzed filling of single cells with dihydrorhodamine, a quantitative sampling technique. The Golgi method revealed the morphology and stratification of cells in detail. The photofilling method allowed us to estimate the frequency of the cell types. From a sample of 243 Golgi-impregnated bipolar cells and 107 photofilled cells, we identified 1 type of rod bipolar cell and 12 types of cone bipolar cells. An analysis based on retinal coverage indicates that this number of types could be contained within the number of bipolar cells known to exist. The dendrites of most cone bipolars contacted all the cones within the individual cone bipolar cell's dendritic field. Types of bipolar cell were encountered at roughly similar frequency, without any one type predominating. The rabbit retina thus contains about a dozen parallel and roughly equipotent through-pathways.

Animals↗

Recoverin negative photoreceptor cells.

Recoverin, a calcium-binding protein, is unique with respect to its cellular regulation. It is present in retinal rods, cones, cone bipolar cells, and in a rare population of cells in the ganglion cell layer. Inappropriate turn-on or turn-off of recoverin expression has been reported both in small cell lung carcinoma cells from patients with cancer-associated retinopathy (Matsusara et al. [1996] Br. J. Cancer 74:1419-1422; Adamus et al. [1998] J. Autoimmun. 11: 523-533; Ohguro et al. [1999] Invest. Ophthalmol. Vis. Sci. 40:82-89) and in cultured retinal neurons (McGinnis et al. [1999] J. Neursci. Res. 55:252-260). In a recent report using double labeling immunofluorescence microscopy methods with antibodies against either rhodopsin and recoverin or arrestin and recoverin, two unique cell phenotypes, rhodopsin-positive and recoverin-negative, and arrestin-positive, and recoverin-negative were observed in vitro. These two unique cell types could be nonphotoreceptor cells in which rhodopsin and arrestin are inappropriately turned on or they are photoreceptor cells in which the recoverin gene is inappropriately turned off. In this study, multiple antibodies were used to study, on a single-cell basis, whether the photoreceptor cell-specific marker, rhodopsin, is inappropriately expressed in nonphotoreceptor cells in our retinal neuronal culture system. We also examined the hypothesis that the two unique cell phenotypes represent the same population of cells. A triple labeling method has been established to visualize recoverin, rhodopsin, and arrestin protein expression simultaneously in cultured retinal neurons. Our data clearly and directly demonstrate that the previously described unique cell phenotypes are the same population of cells, rod photoreceptors. The existence of recoverin-negative photoreceptors demonstrates that the recoverin gene can be regulated independently of other photoreceptor cell-specific proteins and suggests that this primary cell culture may be useful as a model system for investigating the illicit expression of the recoverin gene in cancer associated retinopathy.

Animals↗

Development of On and Off retinal pathways and retinogeniculate projections.

A fundamental functional feature of the visual system, one recognized in the very first electrophysiological retinal recordings ever made, is that some cells respond to light increments (On cells) while others are activated by light decrements (Off cells). The circuitry underlying On and Off responses in the mature retina have been well-established. In particular, it is known that the dendrites of On- and Off-center retinal ganglion cells (RGCs) stratify in different sublamina of the inner plexiform layer (IPL), where they are innervated by spatially segregated On- and Off-cone bipolar cell inputs. Also, segregated into On and Off sublaminae of the IPL are the processes of starburst amacrine cells. In some species (notably ferret and mink) the retinogeniculate projections are also segregated into sublayers of the dorsal lateral geniculate nucleus (dlgn). The mature organizational features summarized above arise gradually during the course of normal development. Thus, the dendrites of immature RGCs initially ramify throughout the IPL before becoming stratified into On or Off sublamina. This developmental event is regulated by the release of glutamate by developing bipolar cells. Treating the developing retina with the glutamate analog 2-amino-4-phosphonobutyric acid (APB) has been found to prevent the stratification of RGC dendrites. In the mature retina APB binds with mGluR6 receptors expressed by On cone and rod bipolar cells which hyperpolarizes these retinal interneurons and blocks their release of glutamate. The effects of short-term APB treatment are reversible by subsequent normal visual experience, while those of long-term treatment appear to be permanent. At the time that developing RGCs are multistratified they respond to both light onset as well as light offset, suggesting that these neurons are initially functionally innervated by On as well as Off-cone bipolar cells. In the dark-adapted state, On-Off responses of immature multistratified RGCs are completely blocked by APB, while at maturity only On responses are APB-sensitive. This suggests that an APB-resistant Off pathway (possibly from rods to Off-cone bipolar cells) is formed relatively late in development, after RGCs attain their stratified state. In contrast to the activity-regulated refinement of stratified On and Off RGCs, the segregated ingrowth of On- and Off-cone bipolar cells occurs in a highly specific manner, and is not dependent on the presence of either RGCs or cholinergic amacrine cells. It is suggested that the directed ingrowth of bipolar cell axons is guided by molecular cues expressed in the extracellular matrix whose identity is yet to be established. There is also evidence that the later formation of segregated On and Off retinogeniculate projections in the ferret is regulated by an activity-dependent Hebbian type mechanism. Blockade of RGC discharges as well as NMDA receptors in the dlgn perturbs the formation of such segregated inputs. Moreover, On and Off RGCs show distinct correlated firing patterns during the developmental period when the intermingled projections of these neurons are being sorted into sign specific sublayers. Collectively, the available evidence indicates that different developmental mechanisms operate on the different components of retinal and retinogeniculate On and Off pathways to attain the segregated state characteristic of the mature visual system.

Animals↗

Spatial density and immunoreactivity of bipolar cells in the macaque monkey retina.

The anatomical substrates of spatial and color vision in the primate retina are investigated by measuring the immunoreactivity and spatial density of bipolar, amacrine and horizontal cells in the inner nuclear layer of the macaque monkey retina. Bipolar cells can be distinguished from amacrine and horizontal cells by their differential immunoreactivity to antisera against glutamate, glycine, GABA, parvalbumin, calbindin (CaBP D-28K), and the L7 protein from mouse cerebellum. The spatial density of bipolar cells is compared to the densities of photoreceptors and ganglion cells at different retinal eccentricities. In the centralmost 2 mm, cone bipolar cells outnumber ganglion cells by about 1.4:1. The density of cone bipolar cells is thus high enough to allow for input to different (parasol and midget) ganglion cell classes by different (diffuse and midget) bipolar cell classes. The density gradient of cone bipolar cells follows closely that of ganglion cells in central retina but falls less steeply in peripheral retina. This suggests that the convergence of cone signals to the receptive fields of ganglion cells in the peripheral retina occurs in the inner plexiform layer. The density of cone bipolar cells is 2.5-4 times that of cones at all eccentricities studied, implying that cone connectivity to bipolar cells remains constant throughout the retina. Different subgroups of bipolar cells are distinguished by their relative immunoreactivity to the different antisera. All rod and cone bipolar cells show moderate to strong glutamate-like immunoreactivity. The bipolar cells that show weak to moderate GABA-like immunoreactivity are also labeled with the antiserum to the L7 protein and are thus identified as rod bipolar cells. Nearly half of all cone bipolar cells showed glycine-like immunoreactivity. The results suggest that the inhibitory neurotransmitter candidates GABA and glycine are segregated respectively in rod and cone bipolar cell pathways. A diffuse, cone bipolar cell type can be identified by the anti-parvalbumin and the anti-calbindin antisera. All horizontal cells show parvalbumin-like immunoreactivity. Nearly all amacrine cells show GABA-like or glycine-like immunoreactivity; a variety of subpopulations also show immunoreactivity to one or more of the other markers used.

Animals↗

Synaptic clustering of GABA(C) receptor rho-subunits in the rat retina.

Polyclonal antibodies which recognize the rho-subunits of the GABA(C) receptor were applied to sections of the rat retina. Strong punctate immunoreactivity was found in the inner plexiform layer (IPL), which was shown by electron microscopy to represent a clustering of the GABA(C) receptors at synaptic sites. During postnatal development diffuse rho-immunoreactivity was first observed at postnatal day P3. Distinct labelling of bipolar cells appeared at P7 and punctate, synaptic labelling was observed at P10. In order to show that the rho-immunoreactive puncta coincide with the axons of bipolar cells, double immunostainings of retinal sections with an antiserum against syntaxin 3 and with the rho-antiserum were performed. The experiments showed that rho-immunoreactive puncta are preferentially located on the axon terminals of rod and cone bipolar cells. In order to determine whether GABA(C) receptor rho-subunits coassemble with GABA(A) receptor subunits, double-labelling experiments were performed with subunit specific antisera. Punctate, putative synaptic clustering was observed with all antisera applied, however, GABA(C) receptor expressing puncta did not coincide with GABA(A) receptor containing puncta. This suggests that there are no synaptic GABA receptors in the retina in which GABA(A) and GABA(C) receptor subunits are coassembled. Similar double-labelling experiments were also performed to find out whether GABA(C) receptors and glycine receptors are colocalized. They were clustered at different synapses. This suggests that synaptic GABA(C) receptors consist of rho-subunits and are not coassembled with GABA(A)- or glycine-receptor subunits.

Animals↗

Post-receptoral mechanisms of colour vision in New World primates.

Diurnal platyrrhines, both di- and trichromats, have magnocellular (M-) and parvocellular (P-) retinal ganglion cells which are morphologically very similar to those found in catarrhines. Catarrhine central P ganglion cells contact single midget bipolar cells, which contact single cones. Physiological recordings of retinal ganglion cells of dichromatic Cebus monkeys showed very similar cell properties to the catarrhine macaque, except that P ganglion cells lacked colour-opponency. We describe the presence of single-headed midget bipolar cells in the Cebus retina. These midget bipolar cells have axon terminal sizes in the same range as the dendritic tree sizes of P ganglion cells as far as 2 mm of retinal eccentricity. This result supports the view that, as in catarrhines, central P ganglion cells of platyrrhines receive input from single midget bipolar cells which in turn, receive input from single cones. This finding is consistent with the idea that a P pathway with one-to-one connectivity was present in the anthropoid ancestor before the divergence between catarrhines and platyrrhines.

Action Potentials↗

Potentiation of 'on' bipolar cell flash responses by dim background light and cGMP in dogfish retinal slices.

The high sensitivity of the vertebrate visual system results from amplification inherent in phototransduction in rods and from the amplification of rod signals on their synaptic transfer at the first synapse with 'on' bipolar cells. These cells possess a metabotropic glutamate receptor linked via a cGMP cascade to the control of cGMP-activated channels. In the study presented here, we show that very dim background light, isomerising only one rhodopsin in 1 out of 10 rods per second, potentiates 'on' bipolar cell responses to superimposed flashes. Responses to dim flashes, which were undetectable above the noise in the dark, were boosted above the increased noise level induced by the background. This potentiation could be reproduced by elevating cGMP, which increases with light, or by dialysing the cells with a non-hydrolysable cGMP analogue. Inhibition of tyrosine kinase activity also reproduced the effect and induced a speeding up of the rising phase of the flash response, similar to the action of dim background light. Conversely, inhibition of tyrosine phosphatase activity blocked the potentiation. These results suggest that cGMP promotes tyrosine-site dephosphorylation of 'on' bipolar cell cGMP-activated channels, resulting in a rise in the sensitivity to cGMP, as has recently been demonstrated for rod cGMP-activated channels. This constitutes a positive feedback mechanism such that as cGMP increases with light, the sensitivity of the channels to cGMP increases and boosts the signal above background noise. This mechanism would allow stochastic resonance to occur, facilitating single-photon detection when dark-adapted, and may therefore lead to improved discrimination.

Adaptation, Physiological↗

Response characteristics and receptive field widths of on-bipolar cells in the mouse retina.

Voltage-clamp and current-clamp recordings were made from bipolar cells in dark-adapted mouse retinal slices. Light-evoked responses fell into three groups corresponding to the rod bipolar cells, on-cone bipolar cells and off-cone bipolar cells. The morphology of the recorded cells confirmed this classification. Intensity-response relations were well fitted by a Michaelis saturation function with Hill coefficients of 1.15 +/- 0.11 (n = 6) for rod bipolar cells and 2.33 +/- 0.06 (n = 4) for cone inputs onto on-cone bipolar cells. In the absence of antagonists for GABA or glycine receptors, light-evoked synaptic currents for all cells displayed linear current-voltage relations that reversed near 0 mV, indicating that very little inhibition was activated under dark-adapted recording conditions. Saturating light stimuli evoked conductances of 0.81 +/- 0.56 nS (n = 4) in rod bipolar cells and 1.1 +/- 0.8 nS (n = 4) in on-cone bipolar cells. Receptive field widths were estimated by flashing a vertical light bar at various locations along the slice. Rod and on-cone bipolar cells had receptive field widths of 67 +/- 16 micrometer (n = 6) and 43 +/- 7 microm (n = 5), respectively. The maximum spatial resolution of an array of such cone bipolar cells was estimated to be 0.3 cycles deg-1, compared with a maximum resolution of 0.5 cycles deg-1 obtained from behavioural studies in mice. Our results suggest that this limit to spatial resolution could be imposed early in the visual system by the size of the bipolar cell receptive fields.

Animals↗

The nuclear receptor transcription factor, retinoid-related orphan receptor beta, regulates retinal progenitor proliferation.

We examined the role of retinoid-related orphan receptor (ROR)beta, a member of the nuclear receptor family of transcription factors, in retinal neurogenesis. In situ hybridization studies showed that RORbeta is expressed in retinal progenitor cells in the embryonic rat retina. Further studies demonstrated that RORbeta colocalizes with Chx10, a transcription factor thought to influence retinal progenitor proliferation (Burmeister, M., Novak, J., Liang, M-Y., Basu, S., Ploder, L., Hawes, N.L. Vidgen, D., Hoover, F., Goldman, D. , Kalnins, V.I., Roderick, T.H., Taylor, B.A., Hankin, M.H. and McInnes, R.R., 1996. Ocular retardation mouse caused by Chx10 homeobox null allele: impaired retinal progenitor proliferation and bipolar cell differentiation. Nature Genetics 12, 376-383). Northern analysis reveals that RORbeta expression is dramatically decreased in the ocular retardationJ mutant, which possesses a defect in the Chx10 gene. Overexpression of RORbeta in retinal progenitors by biolistic transfection results in an increase in the number of large cell clones. These data support a role for RORbeta in regulating retinal progenitor proliferation, possibly via the Chx10 gene.

Animals↗

Capacitance measurements in the mouse rod bipolar cell identify a pool of releasable synaptic vesicles.

The mouse is an important model system for understanding the molecular basis of neuronal signaling and diseases of synaptic communication. However, the best-characterized retinal ribbon-style synapses are those of nonmammalian vertebrates. To remedy this situation, we asked whether it would be feasible to track synaptic vesicle dynamics in the isolated mouse rod bipolar cell using time-resolved capacitance measurements. The results demonstrate that membrane depolarization triggered an increase in membrane capacitance that was Ca(2+) dependent and restricted to the synaptic compartment, consistent with exocytosis. The amplitude of the capacitance response recorded from the easily accessible soma of an intact mouse rod bipolar cell was identical to that recorded directly from the small synaptic terminal, suggesting that in the carefully selected cohort of cells presented here, axonal resistance was not a significant barrier to current flow. This supposition was supported by the analysis of passive membrane properties and a comparison of membrane capacitance measurements in cells with and without synaptic terminals and reinforced by the lack of an effect of sine-wave frequency (200-1,600 Hz) on the measured capacitance increase. The magnitude of the capacitance response increased with Ca(2+) entry until a plateau was reached at a spatially averaged intraterminal calcium of about 600 nM. We interpret this plateau, nominally 30 fF, as corresponding to a releasable pool of synaptic vesicles. The robustness of this measure suggests that capacitance measurements may be used in the mouse rod bipolar cell to compare pool size across treatment conditions.

Animals↗

Protective effect of crocin against blue light- and white light-mediated photoreceptor cell death in bovine and primate retinal primary cell culture.

PURPOSE: The present study was performed to investigate the effect of crocin on blue light- and white light-induced rod and cone death in primary retinal cell cultures. METHODS: Primary retinal cell cultures were prepared from primate and bovine retinas. Fifteen-day-old cultures were exposed to blue actinic light or to white fluorescent light for 24 hours. Cultures were treated by the addition of different concentrations of crocin for 24 hours before light exposure or for 8 hours after light exposure. Cultures kept in the dark were used as controls. Green nucleic acid stain assay was used to evaluate cell death. Rods and cones were immunolabeled with specific antibodies and counted. TUNEL labeling was used to detect fragmented DNA in fixed cells after light exposure. RESULTS: Primary retinal cell cultures contained a mixture of retinal cells enriched in photoreceptors, bipolar cells, and Müller cells. Twenty-four-hour exposure to blue and white light induced death in 70% to 80% of the photoreceptors in bovine and primate retinal cell cultures. Crocin protected the photoreceptors against blue light- or white light-mediated damage in a concentration-dependent manner with an EC50 of approximately 30 microM. TUNEL assays confirmed that crocin protected photoreceptors from light damage. CONCLUSIONS: These results show that blue and white light selectively induce rod and cone cell death in an in vitro model. Crocin protects retinal photoreceptors against light-induced cell death.

Animals↗

Epidermal growth factor is a neuronal differentiation factor for retinal stem cells in vitro.

Stem cells are a tool for in vitro elucidation of the putative role of factors on cell fate. Herein we analyze the role of epidermal growth factor (EGF) on progeny derived from retinal stem cells (RSCs). We isolated cells from neuroretinas of neonate mice. All the proliferating cells harbored the radial glia marker RC2, expressed transcription factors usually found in radial glia (Mash1, Pax6), and met the criteria of stem cells: high capacity of expansion, maintenance of an undifferentiated state, and multipotency demonstrated by clonal analysis. We analyzed the differentiation 7 days after transfer of the cells in different culture media. In absence of serum, EGF led to the expression of the neuronal marker beta-tubulin-III and acquisition of neuronal morphology in 15% of the cells. Analysis of cell proliferation by bromodeoxyuridine incorporation revealed that EGF mainly induced the formation of neurons without stimulating cell cycle progression. Moreover, a pulse of 2-hour EGF stimulation was sufficient to induce neuronal differentiation. Some neurons were committed to the retinal ganglion cell (RGC) phenotype, as revealed by the expression of retinal ganglion markers (Ath5, Brn3b, and melanopsin) and in a few cases to other retinal phenotypes (photoreceptors [PRs] and bipolar cells). We confirmed that the late RSCs were not restricted over time and that they conserved their multipotency by generating retinal phenotypes that usually appear at early (RGC) or late (PRs) developmental stages. Our results show that EGF is not only a factor controlling glial development, as previously shown, but also a potent differentiation factor for retinal neurons, at least in vitro.

Animals↗

Age-related distribution of cones and ON-bipolar cells in the rd mouse retina.

PURPOSE: Retinal dystrophic (rd) mice lose most of their rod photoreceptors within the first three weeks after birth. We determined the age-related distribution of peanut agglutinin lectin (PNA)-labeled cones during the first 12 months of age. We also investigated whether the density of ON-bipolar cells expressing L7 protein was affected by their loss of photoreceptor inputs. METHODS: rd mice were selected from a transgenic strain which expresses an L7-beta-galactosidase fusion gene localized to ON-bipolar cells. Cones were stained with PNA and ON-bipolar cells with bluo-gal (halogenated indolyl-beta-D-galactoside). Retinas were flat-mounted and observed at 1, 2, 3, 6 and 12 months of age. RESULTS: PNA-labeled cones are distributed unevenly across the retina at 1 month postnatal. Their concentration decreases first in the central and far peripheral retina, leaving a ring of labeled cells in the midperipheral region. At 3 months, a larger patch of cones remains in the supero-temporal midperipheral region and a smaller patch in the infero-nasal retina. By 6 months, few cones remain in the infero-nasal retina; by 1 year approximately 100 cones remain in the entire retina, localized to the superior midperipheral region. ON-bipolar cells appear evenly distributed at 1 month. By 2-3 months, relatively more bluo-gal staining is seen in the midperipheral regions underlying dense cone populations. At 6-12 months, bluo-gal label is distributed in a spotty pattern with little or no staining seen in areas of apparent neovascularization. CONCLUSIONS: (1) PNA-labeling of cones in the rd retina deteriorates in a distinct spatial pattern with the longest cone survival in the midperipheral superior retina. (2) ON-bipolar cells are more densely labeled in regions of high cone density during the early months of cone degeneration and, in later stages, show relative decreases in regions of apparent neovascularization.

Age Factors↗

Excitatory synaptic transmission in the inner retina: paired recordings of bipolar cells and neurons of the ganglion cell layer.

Properties of glutamatergic synaptic transmission were investigated by simultaneously voltage-clamping a pair of connected bipolar cells and cells in the ganglion cell layer (GLCs) in the newt retinal slice preparation. Activation of the Ca2+ current in a single bipolar cell was essential for evoking the glutamatergic postsynaptic current in the GLC. Depolarization for as short as 15 msec activated both NMDA and non-NMDA receptors. On the other hand, analysis of the spontaneous glutamatergic synaptic currents of GLCs revealed that these currents consisted of mainly non-NMDA receptor activation with little contribution from NMDA receptors. This suggests that non-NMDA receptors of GLCs are clustered in postsynaptic membrane regions immediately beneath the release sites of bipolar cells and that NMDA receptors have lower accessibility to the released transmitter than non-NMDA receptors. Glutamate that is spilled over from the release sites may activate the NMDA receptors. When a prolonged depolarizing pulse was applied to a bipolar cell, the response induced by non-NMDA receptors was limited greatly by their fast desensitization, whereas NMDA receptors were able to produce a maintained response. The relationship between the pulse duration applied to the bipolar cell and the integrated charge of the response evoked in the GLC was almost linear. Therefore, we propose that both non-NMDA and NMDA receptors cooperate to transfer the graded photoresponses of bipolar cells proportionally to GLCs.

Animals↗

Creatine transporter localization in developing and adult retina: importance of creatine to retinal function.

Creatine and phosphocreatine are required to maintain ATP needed for normal retinal function and development. The aim of the present study was to determine the distribution of the creatine transporter (CRT) to gain insight to how creatine is transported into the retina. An affinity-purified antibody raised against the CRT was applied to adult vertebrate retinas and to mouse retina during development. Confocal microscopy was used to identify the localization pattern as well as co-localization patterns with a range of retinal neurochemical markers. Strong labeling of the CRT was seen in the photoreceptor inner segments in all species studied and labeling of a variety of inner neuronal cells (amacrine, bipolar, and ganglion cells), the retinal nerve fibers and sites of creatine transport into the retina (retinal pigment epithelium, inner retinal blood vessels, and perivascular astrocytes). The CRT was not expressed in Müller cells of any of the species studied. The lack of labeling of Müller cells suggests that neurons are independent of this glial cell in accumulating creatine. During mouse retinal development, expression of the CRT progressively increased throughout the retina until approximately postnatal day 10, with a subsequent decrease. Comparison of the distribution patterns of the CRT in vascular and avascular vertebrate retinas and studies of the mouse retina during development indicate that creatine and phosphocreatine are important for ATP homeostasis.

Animals↗

Functional correlates of somatostatin receptor 2 overexpression in the retina of mice with genetic deletion of somatostatin receptor 1.

Somatostatin-14 (SRIF) and its receptors (sst(1-5)) are found in the mammalian retina. However, scarce information is available on the role of the somatostatinergic system in retinal physiology. We have recently used gene-knockout technology to gain insights into the function of sst(1) and sst(2) receptors in the mouse retina. The sst(1) receptor localizes to SRIF-containing amacrine cells, whereas the sst(2) receptor localizes to several retinal cell populations including rod bipolar cells (RBCs). Molecular data indicate that, in retinas with deletion of the sst(1) receptor (sst(1) KO), sst(2) receptors become overexpressed in concomitance with an increased level of retinal SRIF. To test whether this up-regulation of sst(2) receptors correlates with altered sst(2) receptor physiology, we studied the effect of sst(2) receptor activation on potassium current (I(K)) in isolated RBCs and glutamate release in retina explants. Both I(K) and glutamate release are known to be negatively modulated by sst(2) receptors in the mammalian retina. We used octreotide, a SRIF analogue, to activate selectively sst(2) receptors. Patch-clamp recordings from isolated RBCs indicated that the sst(2) receptor-mediated inhibition of I(K) was significantly larger in sst(1) KO than in control retinas. In addition, HPLC measurements of glutamate release in sst(1) KO retinal explants demonstrated that the sst(2) receptor-mediated inhibition of K(+)-evoked glutamate release was also significantly larger than in control retinas. As a whole, these findings indicate that the overexpression of sst(2) receptors in sst(1) KO retinas can be correlated to an enhanced function of sst(2) receptors. The level of expression of sst(2) receptors may therefore represent a key step in the regulation of sst(2) receptor-mediated responses, at least in the retina.

Animals↗

Immunocytochemical localization of prostaglandin E2 receptor subtypes in porcine ocular tissues. II. Nonuveal tissues.

The localization of PGE2 receptor subtypes (EP1, EP2 and EP3) in nonuveal tissues of porcine eye was investigated by immunocytochemical methods with polyclonal antibodies directed against conserved peptide sequences in the carboxyl terminal region of the receptor subtypes. EP1 was found only in corneal epithelium, while EP2 immunoreactivity was associated with corneal epithelium and endothelium and lens epithelium. EP3 receptor subtype was localized to all ocular tissues examined (except lens fiber cells), i.e. corneal epithelium and endothelium, lens epithelium, retinal photoreceptor and pigmented epithelium, Bipolar cell, horizontal cell, amacrine cell, ganglion cell and Muller cell.

Animals↗