Immortalized hypothalamic luteinizing hormone-releasing hormone neurons express a connexin 26-like protein and display functional gap junction coupling assayed by fluorescence recovery after photobleaching.
Expression of gap junction proteins was studied in the LHRH neuronal cell line, GT1-7, as a first step in defining the signalling mechanisms responsible for the pulsatile secretion of LHRH. GT1-7 cells were found to express a connexin 26-like protein that comigrated with mouse liver connexin 26 and that reacted with connexin 26-specific antibodies on Western blots. Immunofluorescent staining revealed punctate staining in a fraction of the cells, often present at points of apparent contact with neighboring cell bodies or processes. Fluorescence recovery after photobleaching analysis of 5,6-carboxyfluorescein loaded GT1-7 cells showed dye coupling among 20-30% of cells that made contact with other cells, suggesting the presence of functional gap junctions in this cell line.
Matesic
D F
DF
Department of Pediatrics and Human Development, Michigan State University, East Lansing 48824.
Germak
J A
JA
Dupont
E
E
Madhukar
B V
BV
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D000072259:Connexin 26; D017630:Connexins / Q000096:biosynthesis*; D005453:Fluorescence; D005455:Fluorescent Antibody Technique; D017629:Gap Junctions / Q000502:physiology*; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism*; D007031:Hypothalamus / Q000166:cytology / Q000378:metabolism*; D007150:Immunohistochemistry; D051379:Mice; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009474:Neurons / Q000378:metabolism*; D010452:Peptide Biosynthesis; D010455:Peptides / Q000276:immunology
D017630:Connexins; C000607043:Gjb2 protein, mouse; D009419:Nerve Tissue Proteins; D010455:Peptides; D000072259:Connexin 26; D007987:Gonadotropin-Releasing Hormone; D000469:Alkaline Phosphatase