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BOR - Papers in Press, published online ahead of print January 22, 2003.
Biol Reprod 2003, 10.1095/biolreprod.102.010454
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BIOLOGY OF REPRODUCTION 68, 2135–2141 (2003)
DOI: 10.1095/biolreprod.102.010454
© 2003 by the Society for the Study of Reproduction, Inc.

Pregnancy

Down-Regulation of the {alpha}- and ß-Subunits of the Calcium-Activated Potassium Channel in Human Myometrium with Parturition1

B. Matharoo-Ball3, M.L.J. Ashford4, S. Arulkumaran5, and R.N. Khan2,3

Academic Division of Obstetrics & Gynaecology,3 University of Nottingham, Derby City General Hospital, Derby DE22 3NE, United Kingdom Department of Pharmacology & Neuroscience,4 Ninewells Hospital Medical School, University of Dundee, Dundee DD1 9SY, United Kingdom Department of Obstetrics & Gynaecology,5 St. George's Hospital Medical School, London SW17 0RE, United Kingdom

Large-conductance, calcium-dependent potassium (BKCa) channels are implicated in maintaining uterine quiescence during pregnancy. The mechanisms whereby calcium sensitivity of the BKCa channel is dramatically removed at parturition remain unknown. The aim of the present study was to investigate whether this loss of calcium sensitivity of the BKCa channel with the onset of labor is associated with changes in the protein expression of the {alpha}- and/or ß-subunit or arises from a physical dissociation of the {alpha}-subunit from the ß-subunit. The ß-subunit is a key determinant of BKCa-channel Ca2+ sensitivity. Western blot analysis, using {alpha}- and ß-subunit-specific antibodies, detected bands of 110–125 and 36 kDa, respectively. Protein expression levels of the {alpha}-subunit in term labor myometrium were significantly reduced compared with term pregnancy without labor. Furthermore, {alpha}-subunit levels at term pregnancy were significantly increased relative to the nonpregnant state, whereas levels at preterm gestations were unchanged. Densitometric analysis demonstrated significantly decreased ß-subunit levels in term and preterm labor samples compared with term nonlabor samples. Immunoprecipitation studies revealed the presence of both the {alpha}- and ß-subunits in samples taken before or after the onset of labor. We conclude that during labor, the {alpha}-subunit is not physically uncoupled from the ß-subunit, but a decline occurs in the level of ß-subunit protein, which may underlie the loss of calcium and voltage sensitivity of the BKCa channel with labor. Furthermore, reduced ß-subunit protein in preterm labor myometrium implies that ion channels may also contribute to pathophysiological labor.

1 Funded by Tommy's, the Baby Charity, registered charity 1060508

2 Correspondence: Raheela N. Khan, Academic Division of Obstetrics & Gynaecology, Clinical Sciences Building, University of Nottingham, Derby City General Hospital, Derby DE22 3NE, United Kingdom. FAX: 1332 625634; raheela.khan{at}nottingham.ac.uk




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