An attempt at localizing adenylate cyclase in rat calvaria. Influence of sodium fluoride and parathyroid hormone.
To show adenylate cyclase (AC) activity in rat calvaria, it is necessary first to decalcify the specimen. In hard tissues, several enzymes (adenosine triphosphatase (ATPase), alkaline phosphatase (APase), adenylate cyclase (AC) and perhaps pyrophosphatase (PPiase) are able to degrade adenosine triphosphate (ATP). The presence of sodium fluoride (NaF) in the incubation medium reduces the quantity of precipitate formed, compared to that observed using a NaF-free incubation medium. Levamisole, used under the same conditions, gives similar results. Possibly NaF inhibits pyrophosphohydrolase and/or phosphatases which mask the AC activity. Adenylylimidophosphate (AMP-PNP), which is a specific AC substrate, confirms the results obtained with ATP. AC activity is demonstrated cytochemically in the osteoblast and preosteoblast membranes, at the junction between two osteoblasts and along the cytoplasmic processes of the osteoblast which penetrate into the osteoid matrix. The osteocytes never show a precipitate, except those which present some osteoblastic features and then only on the membrane facing the osteogenic layer. An intracellular reaction is also evident and is discussed. Parathyroid hormone (PTH) does not reveal new sites of AC activity but increases the quantity of precipitate observed.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000032:analysis*; D000818:Animals; D000831:Animals, Newborn / Q000033:anatomy & histology*; D003648:Decalcification Technique; D005459:Fluorides / Q000494:pharmacology*; D007978:Levamisole / Q000494:pharmacology*; D010006:Osteoblasts / Q000201:enzymology*; D010281:Parathyroid Hormone / Q000494:pharmacology*; D051381:Rats; D012886:Skull / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D012969:Sodium Fluoride / Q000494:pharmacology*
D010281:Parathyroid Hormone; D007978:Levamisole; D000255:Adenosine Triphosphate; D012969:Sodium Fluoride; D000262:Adenylyl Cyclases; D005459:Fluorides