Adenylate cyclase cytochemistry: a methodological evaluation.
A cytochemical method for the demonstration of adenylate cyclase activity has been evaluated. Enzyme activity in the epithelium of the Müllerian part of the vaginal anlage in neonatal, oestradiol-treated mice has been studied under different experimental conditions. The effects of fixation, incubation conditions and post-fixation have been studied. Variations in the amount of impurities and acid content of the glutaraldehyde do not seem to influence the enzyme activity. High Pb2+ ion concentration seems to promote unspecific staining. Under standard conditions [2 mM Pb (NO3)2' pH 7.2, and incubation temperature 30 degrees C], neither non-enzymatic nor nonsubstrate-dependent lead trapping in the tissue could be observed. The possible contribution of other enzymes utilizing ATP and AMP-P(NH)P as the formation precipitate, has been evaluated. Both ATP and AMP-P(NH)P have been used as substrates in this study. Provided the appropriate control experiments are performed, this cytochemical method is reliable for demonstration of adenylate cyclase activity.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000266:Adenylyl Imidodiphosphate / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn / Q000502:physiology; D002584:Cervix Uteri / Q000201:enzymology*; D004848:Epithelium / Q000201:enzymology; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D006651:Histocytochemistry / Q000379:methods; D007854:Lead / Q000378:metabolism; D051379:Mice; D014621:Vagina / Q000201:enzymology*
D000266:Adenylyl Imidodiphosphate; D007854:Lead; D004958:Estradiol; D000255:Adenosine Triphosphate; D000262:Adenylyl Cyclases