Effect of 17beta-estradiol on mRNA expression of large- conductance, voltage-dependent, and calcium-activated potassium channel alpha and beta subunits in guinea pig.
Large-conductance, voltage- and calcium-activated potassium (MaxiK) channels play a key role in cell excitability. MaxiK channels are composed of a pore-forming alpha-subunit and a regulatory beta-subunit, of which four (beta1-4) genes have been identified. Previous findings suggested that MaxiK channel activity is regulated by estradiol. However, the underlying mechanisms have remained incompletely documented. Therefore, we used reverse transcriptase polymerase chain reaction to clone four cDNA fragments that were specific to the guinea pig alpha, beta1, beta2, and beta4 genes. Using a sensitive ribonuclease protection assay, we found that the alpha and beta4 mRNAs were the most abundant mRNAs in the brain and pituitary, whereas in the aorta, the alpha-subunit was coexpressed with the beta1-subunit. Moreover, there was a significant upregulation of the alpha- but not the beta1-subunit mRNA and the alpha-subunit protein in the aorta of the estrogenvs oil-treated ovariectomized animals. In specific brain areas including preoptic area, ventral hypothalamus, hippocampus, and amygdala, and in the pituitary, neither the alpha- nor beta4-subunit mRNAs were affected by estrogen. These findings suggest that estrogen may not affect the mRNA expression of MaxiK channels in the brain and pituitary. However, estrogen causes increased expression of MaxiK alpha in the aorta, which may explain some of the cardioprotective effects of estrogen in women.
Department of Physiology and Pharmacology, Oregon Health and Science University, Portland, OR.
Khalid
K
Jamali
Barry R
BR
Naylor
Martin J
MJ
Kelly
Oline K
OK
Rønnekleiv
D000679:Amygdala / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism; D001923:Brain Chemistry / Q000187:drug effects*; D003001:Cloning, Molecular; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D006168:Guinea Pigs; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism; D007031:Hypothalamus / Q000187:drug effects / Q000378:metabolism; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D051038:Large-Conductance Calcium-Activated Potassium Channel beta Subunits; D051036:Large-Conductance Calcium-Activated Potassium Channels; D008297:Male; D023282:Nuclease Protection Assays; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism; D024681:Potassium Channels, Calcium-Activated / Q000096:biosynthesis*; D015347:RNA Probes; D012333:RNA, Messenger / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects
D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D051038:Large-Conductance Calcium-Activated Potassium Channel beta Subunits; D051036:Large-Conductance Calcium-Activated Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D015347:RNA Probes; D012333:RNA, Messenger; D004958:Estradiol