Sodium-Calcium Exchanger in Cultured Human Retinal Pigment Epithelium

Mangini, Nancy J.; Haugh-Scheidt, Laura; Valle, J.; Cragoe, Edward J.; Ripps, Harris; Kennedy, Brian G.

doi:10.1006/exer.1997.0390

Cited by 16 publications

(11 citation statements)

References 55 publications

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“…To our knowledge the present study is the first to examine the localization of this Ca 2+ transporter in the human retina. Additionally, our demonstration of Na + /Ca 2+ exchanger immunoreactivity in RPE in situ and a recent study identifying the presence of a cardiac-type Na + /Ca 2+ exchanger in cultured cells [14] are the first to document its presence in human RPE. Our findings are in accordance with earlier studies presenting evidence for the presence of Na + /Ca 2+ exchanger in bovine and dogfish RPE [4].…”

Section: Discussionmentioning

confidence: 85%

“…This antiserum was produced against the canine cardiac sarcolemmal Na + /Ca 2+ exchanger, but also recognizes brain, smooth muscle and kidney Na + /Ca 2+ exchanger and crossreacts with the protein from several species including human (product literature). Recent studies show that p crossreacts with a cardiactype Na + /Ca 2+ exchanger in cultures of human RPE [14].…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Immunohistochemical localization of Na + /Ca 2+ exchanger in human retina and retinal pigment epithelium

Loeffler¹,

Mangini²

1998

Graefe's Archive for Clinical and Experimental Ophthalmology

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Section: Discussionmentioning

confidence: 85%

Section: Methodsmentioning

confidence: 99%

Immunohistochemical localization of Na + /Ca 2+ exchanger in human retina and retinal pigment epithelium

Loeffler¹,

Mangini²

1998

Graefe's Archive for Clinical and Experimental Ophthalmology

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“…Total RNA was isolated from primary cultures of human adult RPE cells with TRIzol (GIBCO) following the methods of Mangini et al (22). For single-strand cDNA synthesis, 5 g of total RNA from each sample was reverse transcribed with Superscript II reverse transcriptase (GIBCO, Gaithersburg MD) and oligo-dT priming.…”

Section: Methodsmentioning

confidence: 99%

Oxidant and antioxidant modulation of chloride channels expressed in human retinal pigment epithelium

Weng

Godley

Jin

et al. 2002

American Journal of Physiology-Cell Physiology

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little is known about the molecular nature of chloride channels in human adult RPE (haRPE) or the effects of oxidative stress on membrane conductance properties. In the present study, we assessed ClC channel and cystic fibrosis transmembrane conductance regulator (CFTR) expression and membrane chloride conductance properties in haRPE cells. ClC-5, ClC-3, ClC-2, and CFTR mRNA expression was confirmed with RT-PCR analysis, and protein expression was detected with Western blot analysis and immunofluorescence microscopy. Whole cell recordings of primary cultures of haRPE showed an outwardly rectifying chloride current that was inhibited by the oxidant H2O2. The inhibitory effects of H2O2 were reduced in cultured human RPE cells that were incubated with precursors of glutathione synthesis or that were stably transfected to overexpress glutathione S-transferase. These findings indicate a possible role for ClC channels in haRPE cells and suggest possible redox modulation of human RPE chloride conductances.immunocytochemistry; patch clamp; glutathione; glutathione S-transferase RETINAL PIGMENT EPITHELIUM (RPE) comprises part of the blood-retina barrier and functions in several processes that are vital for the preservation of sight. Among the crucial roles of this epithelium is the regulation of the volume and electrolyte composition of the subretinal space. This function is achieved largely by regulated transepithelial transport of chloride from the subretinal space to the choroid with the obligatory movement of water. Disruption of RPE chloride transport can result in the accumulation of fluid in the subretinal space and subsequent retinal detachment (3, 26).Chloride transport across the RPE is mediated in part by chloride channels that are stimulated by calcium and cAMP (Ref. 28; cf. Refs. 13,14). In recent whole cell patch-clamp recordings in SV40-transformed cultured human fetal RPE (hfRPE) cells, we (39) identified an outwardly rectifying chloride current that was stimulated by cAMP but was inhibited by the chloride channel blocker DIDS, acidic bathing solutions, or low concentrations of the oxidative agent H 2 O 2 . The molecular identity of the chloride channel(s) responsible for this current was not determined. However, these cells expressed several candidate chloride channels including cystic fibrosis transmembrane conductance receptor (CFTR) and members of the ClC chloride channel family Ref. 39). At present, little is known about the expression of ClC chloride channels in the intact human RPE.Recent evidence suggests that the ClC family of voltage-gated chloride channels may be crucial for retinal function. At least three members of this family are associated with retinal degenerations in mice. Specifically, transgenic mice that were deficient for ClC-3 (34), ClC-2 (6), or ClC-7 (17) were found to develop retinal degenerations and blindness within weeks after birth. The basis of these degenerations is not presently understood.The finding of ClC channel expression in human fetal cells and the consequences of ...

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“…Fig. 1 Mangini et al, 1997). Additionally, these cells can attain a hexagonal packing array, develop pigmentation and catalyse dome formation (cf.…”

Section: Cell Culturementioning

confidence: 97%

“…No ocular disease was reported for any donor. As described previously Mangini et al, 1997 ;Reising et al, 1996), the RPE-Bruch's membranechoroid complex was dissected into small patches (" 0n2 cm#) that were used to initiate cultures. Stock HRPE cultures were propagated in 25 cm# flasks (Falcon, Primaria) in Eagle's Minimal Essential Medium (with Earl's salts) supplemented with 10 % fetal calf serum, 5 % newborn calf serum, essential and non-essential amino acids, 4 m 1-glutamine, 1 µg ml −" amphotericin B and 10 µg ml −" gentamycin.…”

Section: Cell Culturementioning

confidence: 99%