Investigative ophthalmology & visual science · 2001
Expression of the melanin-concentrating hormone receptor in porcine and human ciliary epithelial cells.
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Hintermann E, Erb C, Talke-Messerer C, Liu R, Tanner H, Flammer J, Eberle A N
Аннотация
PURPOSE: To evaluate whether the receptors for melanin-concentrating hormone (MCH) and its functional antagonist alpha-melanocyte-stimulating hormone (alpha-MSH) are expressed in the ciliary epithelium. Furthermore, to examine whether MCH, a neuropeptide involved in fluid and electrolyte homeostasis, may influence ion flux mediated by Na,K (adenosine triphosphatase)-ATPase in a ciliary epithelial cell line.
METHODS: Expression of MCH receptors (MCH-R) and alpha-MSH receptors (MSH-R) on primary porcine ciliary pigmented epithelial (PE) cells and on a human nonpigmented ciliary epithelial (NPE) cell line, ODM-2 was investigated by radioligand binding studies and reverse transcription-polymerase chain reaction (RT-PCR). The MCH-R was further characterized by photocrosslinking. Influence of MCH on Na, K-ATPase activity was evaluated by an Rb(+) transport assay.
RESULTS: MCH-R expression was observed at both the mRNA and protein levels in PE and NPE cells. In contrast, MSH-Rs were not detectable. At the mRNA level, expression of slc-1 was shown and with crosslinking, a 44-kDa protein was labeled. MCH showed no effect on Na,K-ATPase activity of NPE cells.
CONCLUSIONS: The presence of MCH-R in ciliary epithelial cells of both human and porcine origin but the absence of MSH-Rs indicates that in these cells, MCH and alpha-MSH do not form a functionally antagonistic hormonal pair as they do in several other systems. Although effects of MCH on intestinal water and ion transport have been documented, a direct control of Na,K-ATPase activity was not detected in human NPE cells in vitro.
METHODS: Expression of MCH receptors (MCH-R) and alpha-MSH receptors (MSH-R) on primary porcine ciliary pigmented epithelial (PE) cells and on a human nonpigmented ciliary epithelial (NPE) cell line, ODM-2 was investigated by radioligand binding studies and reverse transcription-polymerase chain reaction (RT-PCR). The MCH-R was further characterized by photocrosslinking. Influence of MCH on Na, K-ATPase activity was evaluated by an Rb(+) transport assay.
RESULTS: MCH-R expression was observed at both the mRNA and protein levels in PE and NPE cells. In contrast, MSH-Rs were not detectable. At the mRNA level, expression of slc-1 was shown and with crosslinking, a 44-kDa protein was labeled. MCH showed no effect on Na,K-ATPase activity of NPE cells.
CONCLUSIONS: The presence of MCH-R in ciliary epithelial cells of both human and porcine origin but the absence of MSH-Rs indicates that in these cells, MCH and alpha-MSH do not form a functionally antagonistic hormonal pair as they do in several other systems. Although effects of MCH on intestinal water and ion transport have been documented, a direct control of Na,K-ATPase activity was not detected in human NPE cells in vitro.
Предметные рубрики MeSH
AnimalsBinding, CompetitiveCell LineCiliary BodyDNA PrimersEye ProteinsGene ExpressionHeterotrimeric GTP-Binding ProteinsHumansPigment Epithelium of EyeRNA, MessengerRadioligand AssayReceptors, Pituitary HormoneReceptors, SomatostatinReverse Transcriptase Polymerase Chain ReactionSodium-Potassium-Exchanging ATPaseSwinealpha-MSH