Topographical mapping of GnRH receptors on dispersed mouse pituitary cells by backscattered electron imaging.
Mouse anterior pituitary cells cultured for 2 days were stimulated with one of three biotinylated-GnRH probes ([biotinyl-Lys6]-[D-Lys6]GnRH, [biotinyl-Ser4]-[D-Lys6]GnRH, [biotinyl-Ser4]-[D-Trp6, des-Gly10]GnRH) in the cold (4 degrees C) for 1 hr. These cells were subsequently fixed and an avidin-gold complex was conjugated to the bound GnRH. After a second fixation, the gold label was silver-enhanced for viewing with a scanning electron microscope. Gonadotropes were identified as a result of the labeling procedure, measured for size, and the number of GnRH receptor sites counted. Gonadotropes ranged from 3 to 13 microns in diameter and contained from 23.2 +/- 3.3 to 338.4 +/- 25.2 sites per cell depending upon the size of the cell and the ligand employed. The methods described should be applicable for studying the topographical distribution of a variety of cellular receptors.
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Department of Anatomy and Neurosciences, University of Texas Medical Branch, Galveston 77550.
Miller
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Parkening
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D000818:Animals; D002478:Cells, Cultured; D005260:Female; D006651:Histocytochemistry / Q000379:methods; D008024:Ligands; D051379:Mice; D008810:Mice, Inbred C57BL; D008855:Microscopy, Electron, Scanning / Q000379:methods; D010903:Pituitary Gland, Anterior / Q000737:chemistry* / Q000166:cytology / Q000648:ultrastructure; D011966:Receptors, LHRH / Q000032:analysis*; D012834:Silver
D008024:Ligands; D011966:Receptors, LHRH; D012834:Silver