Light and electron microscopic immunocytochemical analysis of antibodies directed against GnRH and its precursor in hypothalamic neurons.
A battery of antibodies directed against different portions of the precursor to gonadotropin-releasing hormone (GnRH), as well as to the mature decapeptide, were characterized immunocytochemically in two ways. Absorption experiments were used to determine the epitope recognized by each antiserum. Electron microscopic immunocytochemistry was then used to define the subcellular organelles that contained reaction product when tissue was incubated with these reagents. These latter observations helped to determine if the antibody recognized the epitope as part of the intact precursor or only after it had been cleaved from parent protein. Our results demonstrate that the GnRH precursor is routed from the rough endoplasmic reticulum through the Golgi apparatus to the secretory vesicles. Furthermore, we show that initial cleavage and processing of the GnRH precursor begin in the cell soma. These antibodies should be useful in the future in determining changes in processing of precursor in animals that differ in endocrine function.
Department of Anatomy and Cell Biology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
A J
AJ
Silverman
J W
JW
Witkin
R P
RP
Millar
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D005260:Female; D007031:Hypothalamus / Q000032:analysis* / Q000648:ultrastructure; D007106:Immune Sera / Q000276:immunology; D007150:Immunohistochemistry; D007163:Immunosorbent Techniques; D008297:Male; D008853:Microscopy; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009474:Neurons / Q000032:analysis* / Q000648:ultrastructure; D010906:Pituitary Hormone-Releasing Hormones / Q000032:analysis*; D011498:Protein Precursors / Q000032:analysis*; D011499:Protein Processing, Post-Translational; D051381:Rats; D011916:Rats, Inbred F344
D007106:Immune Sera; D010906:Pituitary Hormone-Releasing Hormones; D011498:Protein Precursors