TY - JOUR
T1 - Differential expression of adenylyl cyclases in the rat nephron
AU - Bek, Martin J.
AU - Zheng, Shaopeng
AU - Xu, Jing
AU - Yamaguchi, Ikuyo
AU - Asico, Laureano D.
AU - Sun, Xiao Guang
AU - Jose, Pedro A.
N1 - Funding Information:
This work was supported by grants from the National Institutes of Health, HL 23081, DK 52612, DK 39308, HL 58536, and a fellowship grant from the National Kidney Foundation (MB). We thank Dr. Richard T. Premont (Department of Medicine, Duke University Medical Center, Durham, NC, USA) for the generous gift of the AC IX antibody. We are indebted to Dr. Peter Andrews (Department of Cell Biology, Georgetown University Medical Center, Washington D.C., USA) for his advice and thorough review of the immunohistochemical stainings. We wish to thank Mr. Matt Higgs for reviewing our manuscript.
PY - 2001
Y1 - 2001
N2 - 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 α3i, αs and β/α 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 VIImRNA 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. Conclusion. Knowledge of the distribution of ACs may help establish the linkage between receptors and specific AC isoforms and define their functions.
AB - 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 α3i, αs and β/α 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 VIImRNA 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. Conclusion. Knowledge of the distribution of ACs may help establish the linkage between receptors and specific AC isoforms and define their functions.
KW - Cell regulation
KW - Nephron
KW - Renal AC isoforms
KW - Second messenger cAMP
KW - Signal transduction
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U2 - 10.1046/j.1523-1755.2001.060003890.x
DO - 10.1046/j.1523-1755.2001.060003890.x
M3 - Article
C2 - 11532084
AN - SCOPUS:0035720554
VL - 60
SP - 890
EP - 899
JO - Kidney International
JF - Kidney International
SN - 0085-2538
IS - 3
ER -