Differential expression of adenylyl cyclases in the rat nephron

• Published in: Kidney international Vol. 60; no. 3; pp. 890 - 899
• Main Authors: Bek, Martin J., Zheng, Shaopeng, Xu, Jing, Yamaguchi, Ikuyo, Asico, Laureano D., Sun, Xiaoguang, Jose, Pedro A.
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01-09-2001; Nature Publishing; Elsevier Limited
• Subjects: Adenylyl Cyclases - biosynthesis; Adenylyl Cyclases - genetics; Animals; Biological and medical sciences; Blotting, Western; cell regulation; Fundamental and applied biological sciences. Psychology; Immunohistochemistry; Isoenzymes - biosynthesis; Male; nephron; Nephrons - enzymology; Rats; Rats, Inbred WKY; renal AC isoforms; Reverse Transcriptase Polymerase Chain Reaction; RNA - analysis; RNA - biosynthesis; second messenger cAMP; signal transduction; Vertebrates: urinary system; cell regulation; nephron; signal transduction; renal AC isoforms; second messenger cAMP; Animal model; Protein kinase C; Rat; Enzyme; Transferases; Rodentia; Phosphorus-oxygen lyases; Cyclic AMP; Reverse transcription; Isoforming; Lyases; Polymerase chain reaction; Adenylate cyclase; Signal transduction; Vertebrata; Mammalia; Animal; Molecular biology; G protein
• ISSN: 0085-2538; 1523-1755
• DOI: 10.1046/j.1523-1755.2001.060003890.x

Differential expression of adenylyl cyclases in the rat nephron.
Adenylyl cyclases (ACs) are a family of enzymes that catalyze the formation of the second-messenger cyclic adenosine 3′,5′-monophosphate (cAMP). At least nine isoforms of AC have been cloned. These isoforms differ in their tissue distribution and basal activity. AC isoforms also differ in their capacity to be stimulated or inhibited by G protein αi, αs and β;/γ subunits, protein kinase C, and intracellular calcium. The distribution of ACs in the kidney is only partially known, although it is known that ACs play important roles in kidney signal transduction. Several receptors are known to couple to AC, but their linkage to individual AC isoforms in the kidney is not known.
This study investigated the tissue distribution of AC isoforms along the nephron of Wistar-Kyoto rats using reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and immunoblotting.
While AC VI and IX mRNA were found in all nephron segments, there was no expression of AC VIII mRNA. ACs II through V and VII mRNA were variably found in specific nephron segments. mRNA for AC isoforms II, III, VI, VII, and IX were expressed in renal proximal tubules. All of the AC isoforms studied, except VIII, were found in glomeruli. Immunoblotting and immunohistochemistry confirmed the mRNA results. AC isoforms II, III, IV, and IX were expressed in luminal rather than in basolateral membranes. However, immunohistochemical studies were not feasible for the other isoforms that could be expressed in basolateral membranes.
Knowledge of the distribution of ACs may help establish the linkage between receptors and specific AC isoforms and define their functions.