Differential expression of adenylyl cyclases in the rat nephron.
BACKGROUND
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 alpha(i), alpha(s) and beta/gamma 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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
Knowledge of the distribution of ACs may help establish the linkage between receptors and specific AC isoforms and define their functions.
Bek
M J
MJ
Department of Pediatrics, Georgetown University Medical Center, Washington, DC 20007, USA.
Zheng
S
S
Xu
J
J
Yamaguchi
I
I
Asico
L D
LD
Sun
X G
XG
Jose
P A
PA
D000262:Adenylyl Cyclases / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000096:biosynthesis; D008297:Male; D009399:Nephrons / Q000201:enzymology*; D012313:RNA / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D011921:Rats, Inbred WKY; D020133:Reverse Transcriptase Polymerase Chain Reaction
D007527:Isoenzymes; D012313:RNA; D000262:Adenylyl Cyclases