Down-regulation of adrenal neuronal nitric oxide synthase mRNAs and proteins after deoxycorticosterone acetate-salt treatment in rats.
The aim of this study was to evaluate the possible changes of adrenal neuronal nitrite oxide synthase (nNOS) messenger RNA (mRNA) and protein of rats after deoxycorticosterone acetate (DOCA)-salt treatment. We determined adrenal nNOS expression in 12 vehicle-treated and 13 DOCA-salt-treated rats by in situ hybridization, immunohistochemistry, and multiplex RT-PCR methods. Adrenal nNOS was also detected by Western blot in five vehicle-treated and five DOCA-salt-treated rats. The results showed that adrenal nNOS mRNA and nNOS immunoreactivities were mainly localized in the medulla and some in the regions of zona glomerulosa. DOCA-salt treatment inactivated nNOS mRNA and peptide expression prominent in the adrenal medulla and slight in the zona glomerulosa. The relative quantities of nNOS mRNA in the adrenals of the DOCA-salt-treated group was 8.8-fold decreased. At the same time, the relative quantities of steroid acute regulatory protein mRNA and phenylethanolamine N-methyltransferase mRNA in the adrenals of the DOCA-salt-treated group were significantly decreased. Western blots showed that total adrenal nNOS were 3.7-fold down-regulated after DOCA-salt treatment. Our results indicated that the down-regulation of adrenal nNOS synthesis might be associated with the inactivation of adrenal function in face of volume expansion.
Lai
Feng-Jie
FJ
Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Hsin
Ya-Chieh
YC
Huang
Shiue-Chin
SC
Cheng
Ching-Li
CL
Hsin
Shih-Chie
SC
Hsieh
Ming-Chia
MC
Shin
Shyi-Jang
SJ
D000311:Adrenal Glands / Q000201:enzymology / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D003900:Desoxycorticosterone / Q000008:administration & dosage / Q000494:pharmacology*; D004347:Drug Interactions; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism*; D010625:Phenylethanolamine N-Methyltransferase / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012965:Sodium Chloride / Q000008:administration & dosage / Q000494:pharmacology*; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D012333:RNA, Messenger; D003900:Desoxycorticosterone; D012965:Sodium Chloride; D052248:Nitric Oxide Synthase Type I; D014446:Tyrosine 3-Monooxygenase; D010625:Phenylethanolamine N-Methyltransferase