Immunocytochemical demonstration of the binding of growth-related polypeptide hormones on chick embryonic tissues

The presence of endogenous growth-related polypeptide hormones, such as growth hormone (GH), somatomadin-C/insulin-like growth factor-1 (SM-C/IGF-1), prolactin (PRL) and Mullerian inhibiting substance (MIS) on chick embryonic tissues have been detected by electron microscopic (EM) immunocytochemistry. Antiserum against GH, anti-SM-C/IGF-1, anti-PRL and anti-MIS were used respectively as primary antibodies for immunolabeling probes by peroxidase (PO) and avidin-biotin complex (ABC)-gold ligands. Cross-reaction studies by ELISA showed negative or weak antigen-antibody interactions. Chick embryos, gonads, and Mullerian ducts (Mds) of various ages were fixed in 2.5% glutaraldehyde for 30 min. Washes in phosphate buffer were administered between each of the following incubations: (i) 2% BSA; (ii) primary antibody; (iii) biotinylated or PO-conjugated secondary antibody; (iv) avidin conjugated with gold particles. SM-C/IGF-1 bindings were negative on 1d embryonic disc, heavily stained on 2d endoderm. However, the GH bindings were found on the embryonic layers of 1d and 2d embryos, and increasing on the luminal epithelial cells of Mds during development. PRL was found in parallel with GH, but in less amount. The 10d Mds were double labeled with anti-SM-C/IGF-1-gold and anti-MIS-PO (MIS-PO), and the results showed SM-C/IGF-I negative, but MIS-PO positive bindings. This study provides the first immunocytochemical evidences for: (i) The presence of GH, SM-C/IGF-1, PRL and MIS bindings on chick embryonic tissues, and further supports their potential role as growth mediators during embryonic development. (ii) The amount of GH and MIS bindings were found correspondingly to their physiological status of Md growth or regression.(ABSTRACT TRUNCATED AT 250 WORDS)