Expression and cellular distribution of the alpha 1 gap junction gene product in the ocular pigmented ciliary epithelium.
The expression of four different gap junction (GJ) transcripts, corresponding to the alpha 1, alpha 3, beta 1 and beta 2 gene products, has been examined in the ciliary epithelium of human and bovine eyes, and in cultures of ciliary epithelial cells. Northern blot analysis revealed that alpha 1 mRNA, 3.6-kb in size, was the predominant transcript expressed in intact tissue and in cultures of pigmented ciliary epithelial cells (PE). No transcripts from the alpha 3, beta 1 or beta 2 gap junction genes were detected in intact tissue or ciliary epithelial cells as demonstrated by Northern blotting. When the levels of alpha 1 gap junction mRNA were compared between PE and NPE in primary cultures, a striking difference was observed in the level of alpha 1 transcripts: there was about a 6 to 8-fold increase in alpha 1 levels in PE cells, relative to the NPE cells. To verify the differential level of expression of alpha 1 GJ mRNA in the two cell types, indirect immunofluorescence localization studies were performed on semithin cryostat sections of ciliary processes. These studies revealed that alpha 1 gap junctions are present at the apical and lateral borders of PE cells, i.e. at the apical plasma membranes domains of PE-NPE cells, and at the lateral plasma membrane regions of PE-PE cells borders. Further analysis by immunoblotting confirmed that the 43 kD alpha 1 gap junction protein was the major gap junction gene product in the ciliary epithelium.
Coca-Prados
M
M
Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, CT 06510.
Ghosh
S
S
Gilula
N B
NB
Kumar
N M
NM
D000818:Animals; D015152:Blotting, Northern; D002417:Cattle; D002478:Cells, Cultured; D002924:Ciliary Body / Q000378:metabolism*; D017630:Connexins; D015342:DNA Probes; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D010857:Pigment Epithelium of Eye / Q000378:metabolism*; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
D017630:Connexins; D015342:DNA Probes; D008565:Membrane Proteins; D012333:RNA, Messenger; D012313:RNA