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At least 595 records · Page 33Linked to original sources
PubMed1986

Three types of photoreceptors in the pineal and frontal organs of frogs: ultrastructure and opsin immunoreactivity.

The pineal complex in frogs (Rana esculenta, R. temporaria, R. tigrina, R. arvalis) was studied by conventional electron microscopy and postembedding rhodopsin immunoelectron microscopy. Three types of photoreceptor cells were found in both the pineal and frontal organs. In the pineal organ, most of the photoreceptors exhibited rhodopsin-immunoreactive outer segments and large inner segments with a large ellipsoid of densely packed mitochondria ("rod-like" photoreceptors). A small number of photoreceptors was rhodopsin-immunonegative ("cone-like" photoreceptors). In both Rana esculenta and R. temporaria, the latter were either supplied with an oil droplet and an ellipsoid in their inner segment, or they were electron-lucent with a small inner segment without an ellipsoid. In contrast, the frontal organ displayed many immunonegative "cone-like" outer segments and few rhodopsin-immunoreactive "rod-like" photoreceptors. In both organs, the basal processes of the photoreceptor cells were found to form ribbon-containing axonal pedicles which synapsed with the dendrites of secondary neurons. The latter rarely received any further afferences by conventional synapses. The frog pineal organ is considered a predominantly "rod-type" and the frontal organ a "cone-type" photosensory organ. The presence of three kinds of pineal/frontal photoreceptors is discussed in connection with the occurrence of different photopigments (rhodopsin/porphyropsin, iodopsin, ultraviolet and/or blue pigments) enabling the animal to discriminate by the pineal complex environmental light in various ranges of the spectrum.

Animals↗
PubMed2001

Characterization of human GRK7 as a potential cone opsin kinase.

PURPOSE: Homozygous inactivation of the mouse gene for GRK1 (G protein-coupled receptor kinase 1, or rhodopsin kinase) causes severe defects in the recovery of cone phototransduction. However, electroretinographic (ERG) analyses of human oguchi patients with defective GRK1 alleles showed normal or slightly abnormal photopic responses. It remains unclear why the loss of GRK1 yields such different phenotypes in the recovery of mouse and human cones. We examined the localization and enzyme activity of GRK7, the human ortholog of the seventh member of the GRK family, in an attempt to understand its potential role in photopic vision. METHODS: Bioinformatic approaches were used to identify the human GRK7 gene. Human and bovine GRK7 cDNAs were isolated by RT-PCR. Recombinant GRK7, expressed in insect cells, was used to phosphorylate activated rhodopsin. Antibodies raised against GRK7 peptides were used to examine the retina specific expression of GRK7 by immunoblotting and its subcellular localization by immunocytochemistry. RESULTS: The human GRK7 gene is located on chromosome 3q21, spans at least 10 Kb and consists of 4 exons. In human, GRK7 is expressed exclusively in the retina and is found in all retinal neurons, and specifically, in cone outer segments. Recombinant human GRK7 catalyzes rhodopsin phosphorylation in a light dependent manner. We provide evidence that GRK1 and GRK7 are co-expressed in human cones. In contrast, mouse GRK7 is expressed in many tissues including retina where photoreceptors apparently do not express GRK7. CONCLUSIONS: The presence of GRK7 in human, but not in mouse, cone outer segments suggests that GRK7 may function to provide the normal photopic vision reported by oguchi patients with a defective GRK1 gene. The absence of GRK7 expression in cone outer segments of mice is consistent with the notion that mouse cones rely solely on GRK1 to shutoff cone visual pigments.

Amino Acid Sequence↗
PubMed1998

Structure and developmental expression of the mouse RGR opsin gene.

PURPOSE: The aim of this study is to isolate and characterize cDNA clones and the genes that encode mouse RPE retinal G protein-coupled receptor (RGR) and to analyze expression of the RGR gene in the developing mouse retina. The conserved amino acid sequences of RGR from various mammals can be compared to the amino acid sequence motif of G protein-coupled receptors. METHODS: Mouse RGR cDNA and gene clones were isolated from a retina cDNA library and 129SV genomic DNA library, respectively. The expression of RGR in the developing C57BL/6J mouse retina was analyzed by immunohistochemical staining with a polyclonal antipeptide antibody. RESULTS: The deduced amino acid sequence of mouse RGR is 78% and 81% identical to that of bovine and human RGR, respectively. The mouse RGR gene is split into seven exons and extends about 11 kb. Two predominant mRNA transcripts, 1.9 and 1.7 kb in length, and a third, relatively faint, 5.5-kb transcript were detected in mouse eye by hybridization to a RGR cDNA probe. Frozen sections of C57BL/6J mouse retina at various stages of development were incubated with a mouse RGR antipeptide antibody. RGR immunoreactivity was first seen at postnatal day 2 (P2) in centrally located RPE cells. From day P6 to P12, there was an increase in the number and intensity of immunoreactive RPE cells in the central and mid-peripheral regions of the retina, while the most peripheral RPE cells were still negative. By day P16, the length of the RPE monolayer was immunoreactive, and staining of the central RPE cells was markedly more intense than at younger ages. CONCLUSIONS: Mouse and human RGR are highly conserved. A gradient of RGR expression in RPE extends from the central to the peripheral retina during development. In reference to the appearance of melanin-positive differentiated RPE cells, the induction of RGR expression is a relatively late event in the maturation of the retina.

Aging↗
PubMed2005

Differential regulation of melatonin synthesis genes and phototransduction genes in embryonic chicken retina and cultured retinal precursor cells.

PURPOSE: Photoreceptor differentiation involves the activation of two specific sets of genes; those encoding the proteins of the phototransduction cascade and those encoding the enzymes of the melatonin synthesis pathway, arylalkylamine N-acetyltransferase (AANAT) and hydroxyindole O-methyltransferase (HIOMT). The purpose of the present study was to examine the conditions of AANAT and HIOMT gene activation, relative to that of selected phototransduction markers (alpha-transducin and opsins), in both in vivo and in vitro differentiating photoreceptors of the chicken retina. METHODS: Neural retina RNA was obtained between embryonic day 7 (E7) and posthatch day 8 (P8) and analyzed on northern blots with cDNA probes to AANAT, HIOMT, visinin, alpha-transducin, rhodopsin, and the four cone opsins. Cell cultures were prepared from E7 chicken neural retina and incubated for two to four days in vitro, either in basal medium or in serum-supplemented medium or in medium containing an insulin-based supplement. RNA from the cultured cells was analyzed on northern blots as above. Real time RT-PCR was used to confirm in vitro changes in HIOMT and red opsin mRNA levels. The cultured cells were transfected with promoter-reporter plasmids for direct analysis of HIOMT promoter regulation by the dual luciferase method. RESULTS: The different mRNAs composing the photoreceptor phenotype appeared at E7 (visinin), E10 (alpha-transducin), E14 (HIOMT), E15 (rhodopsin, red opsin, and green opsin), E16 (AANAT), E17 (blue opsin), and E18 (violet opsin). In the early differentiating cones of the central retina, HIOMT mRNA appeared two days earlier than red opsin and green opsin mRNAs (E12 rather than E14). In cultured embryonic neural retina cells, basal medium was sufficient to activate alpha-transducin gene transcription, an insulin-based supplement was sufficient to activate HIOMT gene transcription, whereas serum was required for red opsin gene transcription after two days in vitro. All serum batches were able to activate red opsin gene transcription, whereas some of them failed to activate HIOMT gene transcription. Activation of the HIOMT gene promoter by an insulin-based supplement and by serum was confirmed after transfection of chicken embryonic neural retina cells with promoter-reporter plasmids. CONCLUSIONS: Activation of the melatonin synthesis genes in vivo takes place in a time window very close to that of early opsins. However, a 24-48 h lead of HIOMT gene expression over early opsins was clearly observed. Our in vitro experiments indicate that different exogenous signals are required to activate the different genes encoding photoreceptor specific functions. Significantly, marker genes for light sensitivity (red opsin) and for melatonin synthesis (HIOMT) appear to be activated in response to different signals.

Acetylserotonin O-Methyltransferase↗
PubMed1983

Biogenesis of blowfly photoreceptor membranes is regulated by 11-cis-retinal.

Biogenesis of photoreceptor membranes has been investigated by analyzing the rhodopsin and opsin content of microvillar photoreceptor membranes after injecting retinal isomers and radioactive amino acids into the compound eyes of carotenoid-deficient blowflies. The amount of rhodopsin in the membranes was measured photometrically in extracts of isolated rhabdoms. The opsin content of the membranes and the level of radioactive labelling of opsin were measured after separating the membrane proteins by dodecyl sulphate/polyacrylamide gel electrophoresis. In the photoreceptor membrane of carotenoid-deficient flies the rhodopsin and opsin content is less than 4% of that in normal flies where opsin may constitute about 65% of the total membrane protein. In carotenoid-deficient flies the incorporation of rhodopsin into photoreceptor membranes is triggered by 11-cis-retinal but not all-trans retinal. After injection of 11-cis-retinal the opsin content of the photoreceptor membranes increases in parallel with the rhodopsin content. Radioactive labelling of opsin reveals that 11-cis-retinal triggers an incorporation of newly synthesized opsin into photoreceptor membranes whereas all-trans retinal does not induce the assembly of opsin into photoreceptor membranes. Light-dependent incorporation of radioactive labelled opsin into membranes with a high rhodopsin content shows that not only the visual pigment chromophore but also the opsin undergoes a light-dependent turnover. The findings raise the possibility that opsin synthesis in blowfly photoreceptors and consequently the assembly of photoreceptor membranes is regulated by 11-cis-retinal.

Animals↗
PubMed1997

Spatiotemporal coordination of rod and cone photoreceptor differentiation in goldfish retina.

In this study, we have compared spatial and temporal aspects of development of new rods and cones in the adult goldfish by using a combination of bromodeoxyuridine immunocytochemistry and opsin in situ hybridization to determine the intervals between terminal mitosis (cell "birth") and expression of opsin mRNA for each photoreceptor cell type. The goldfish opsins include rod opsin and four different cone opsins: red, green, blue, and ultraviolet. In a cohort of photoreceptors born at the same time, rods expressed opsin mRNA within 3 days of cell birth, while expression of cone opsin mRNA required at least 7 days. This temporal discrepancy in differentiation, coupled with a discordance in the site of cell genesis of rods and cones, allowed opsin expression to commence in both cell types in approximately the same retinal location. Commitment to the generic cone phenotype occurred within approximately 6 days throughout the cone cohort, as indicated by expression of interphotoreceptor retinoid-binding protein (IRBP) mRNA, but expression of a specific spectral phenotype was delayed until rods differentiated nearby. Onset of expression of cone opsin mRNA followed a phenotype-specific sequence: red, then green, then blue, and finally ultraviolet; in situ hybridization with two opsin probes confirmed that individual photoreceptors expressed only one type of opsin as they differentiated. This stepwise process of cone differentiation is consistent with the hypothesis that cell-cell interactions among developing photoreceptors may coordinate selection of specific photoreceptor phenotypes.

Animals↗
PubMed2004

Unusual cone and rod properties in subterranean African mole-rats (Rodentia, Bathyergidae).

We have determined the presence of spectral cone types, and the population densities of cones and rods, in subterranean mole-rats of the rodent family Bathyergidae, for which light and vision seems of little importance. Most mammals have two spectral cone types, a majority of middle- to long-wave-sensitive (L-) cones, and a minority of short-wave-sensitive (S-)cones. We were interested to see whether the subterranean bathyergids show the same pattern. In three species, Ansell's mole-rat Cryptomys anselli, the giant mole-rat Cryptomys mechowi and the naked mole-rat Heterocephalus glaber, spectral cone types and rods were assessed immunocytochemically with opsin-specific antibodies. All three species had rod-dominated retinae but possessed significant cone populations. A quantitative assessment in C. anselli and C. mechowi revealed surprisingly low photoreceptor densities of 100 000-150 000/mm(2), and high cone proportions, approximately 10% (8000-15 000/mm(2)). In all three species, the vast majority of the cones were strongly S-opsin-immunoreactive; L-opsin immunoreactivity was much fainter. In C. anselli, approximately 20% of the cones showed exclusive S-opsin label, approximately 10% exclusive L-opsin label and approximately 70% strong S-opsin and faint L-opsin double label (potential dual-pigment cones). This is the first observation in any mammal of an S-opsin dominance and low levels of L-opsin across the entire retina. It contrasts starkly with the situation in the muroid blind mole-rat Spalax ehrenbergi, which has been reported to possess L-opsin but no S-opsin. Evidently, within rodents an adaptation to subterranean life is compatible with very different spectral cone properties.

Animals↗