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Tissue-Specific Protein Kinase C Isoform Expression in Rat Uterine Tissue

Trevania Saunders

Eric Bieber, MD

Department of Obstetrics and Gynecology, University of Chicago. Chicago, Illinois

Mark Phillippe, MD

Department of Obstetrics and Gynecology, University of Chicago, Chicago, Illinois; mphillip{at}babies.bsd.uchicago.edu

Objective: Activation of the phosphatidylinositol signaling pathway plays a key role during the generation of agonist-stimulated phasic myometrial contractions. Protein kinase C (PKC), a component of this signaling pathway, has been previously shown to produce feedback inhibition of agonist-stimulated myometrial contractions. The studies described in this report were performed to survey the tissue-specific expression of several PKC isoforms in the rat uterus.

Methods: Uterine tissue was obtained from timed pregnant and normally cycling adult female Sprague-Dawley rats. Immunohistochemical studies were performed using the Vectastian ABC immunostaining technique and PKC isoform-specific polyclonal antibodies. Western blot studies were peformed using myometrial tissue separated into cytosol and membrane fractions by differential centrifugation.

Results: These studies confirmed significant expression of the PKC-, -ß₂, -, -, and - isoforms in myometrium from pregnant and estrus rats, whereas only trace or no expression of the PKC-ß₁, -, -, and - isoforms was observed. Expression of the PKC-, -ß₂, and - isoforms decreased modestly during the latter days of gestation; in contrast, PKC- and - remained stable during this period. The immunohistochemical studies confirmed expression of the PKC-, -ß₂, -, -, and - isoforms in both circular and longitudinal smooth-muscle layers of the near-term pregnant rat uterus.

Conclusion: In summary, these studies have confirmed significant levels of expression of several isoforms of PKC in estrus and near-term pregnant rat uterine tissue, which was most prominent in the smooth-muscle cells of the myometrium.

Key Words: Immunohistochemistry • myometrium • phosphatidylinositol signaling pathway • protein kinase C • rat uterus

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