Adenylate cyclase cytochemistry: a methodological evaluation |
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Authors: | Stener Kvinnsland |
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Affiliation: | (1) Institute of Anatomy, University of Bergen, Bergen, Norway |
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Abstract: | Synopsis 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°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. |
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