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Hypertension. 1995;25:350-355

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(Hypertension. 1995;25:350-355.)
© 1995 American Heart Association, Inc.


Articles

Testosterone Regulation of Renal {alpha}2B-Adrenergic Receptor mRNA Levels

Guodong Gong; Mark L. Johnson; William A. Pettinger

From the Department of Medicine, Creighton University Medical Center, Omaha, Neb.

Correspondence to Mark L. Johnson, PhD, Department of Medicine, Creighton University Medical Center, 601 N 30th St, Suite 6730, Omaha, NE 68131.

Abstract Androgens regulate blood pressure and renal {alpha}2-adrenergic receptors in a parallel fashion in the spontaneously hypertensive rat (SHR). The present studies investigated whether this regulation of renal {alpha}2B-adrenergic receptors occurs at the mRNA level. Male and female SHR were gonadectomized at 4 weeks of age. The gonadectomized rats were implanted with or without testosterone propionate. Sham- gonadectomized rats served as controls. Total kidney RNA was purified, and {alpha}2B-adrenergic receptor mRNA was quantified with a reverse transcriptase–polymerase chain reaction (RT-PCR) assay. The assay uses a mimic RNA added at known concentrations to the sample RNA. The mimic was constructed from the target sequence in the {alpha}2B-adrenergic receptor mRNA plus a 20-bp insertion of a random nucleotide sequence. The amount of {alpha}2B-adrenergic receptor mRNA present in each sample was obtained by determining the equivalence point between the amount of RT-PCR product formed in the target band versus the mimic band, which were resolved by gel electrophoresis. Intact males had more than two times as much {alpha}2B-adrenergic receptor mRNA as intact females. Castration of males reduced the male-female difference by more than 60%. Ovariectomy slightly increased the {alpha}2B-adrenergic receptor mRNA level compared with that of intact females. Treatment with testosterone elevated {alpha}2B-adrenergic receptor mRNA levels of gonadectomized males and females to the level of intact males. The {alpha}2B-adrenergic receptor mRNA levels correlated remarkably well with renal {alpha}2-adrenergic receptor density. We conclude that testosterone regulates renal {alpha}2B-adrenergic receptor gene expression at the mRNA level in the SHR.


Key Words: RNA, messenger • receptor, adrenergic, alpha • androgens • testosterone




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