M F Thomure1, M J Gast, N Srivastava, R M Payne. 1. Department of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Abstract
OBJECTIVE: Creatine kinase (CK) isoenzymes play an important role in cellular energy transduction. Two isoenzymes of creatine kinase, ubiquitous mitochondrial creatine kinase (uMtCK) and cytosolic brain creatine kinase (BCK), are postulated to form the creatine phosphate (CP) shuttle, in which creatine serves to transport high-energy phosphate from the mitochondria to its site of utilization. Coordinate regulation of these genes is essential for efficient energy transduction. We examined human CK isoenzyme regulation in placentas during all three trimesters of gestation to define the mRNA and protein expression patterns of uMtCK and BCK and to test the CP shuttle hypothesis. METHODS: Placental samples were collected from a total of 26 patients from the first, second, and third trimesters. Total RNA and protein were prepared from each sample and quantified. Quantitative RNA analysis was performed by gel electrophoresis and dot blot techniques using isoenzyme-specific human cDNA probes for uMtCK and BCK. Protein expression of uMtCK and BCK was examined by Western blot analysis using isoenzyme-specific antibodies to uMtCK and BCK. RESULTS: Analysis of RNA demonstrated the coordinate expression of uMtCK and BCK mRNAs in human placenta, with peak expression of both in the term placentas. Western blot analysis demonstrated coordinate expression of uMtCK and BCK proteins in the first and second trimesters, but not in the term placenta. Expression levels of uMtCK and BCK proteins were not consistent with their respective mRNA levels in the term placenta. CONCLUSION: Expression of uMtCK and BCK in human placenta is highly regulated, and post-transcriptional regulation of uMtCK and BCK expression occurs in the term placenta. The coordinate regulation of uMtCK and BCK in human placenta supports the CP shuttle hypothesis. This analysis demonstrates that human placenta has high energy needs that can change rapidly; thus, a functioning CP shuttle may be important in the maintenance and termination of pregnancy.
OBJECTIVE:Creatine kinase (CK) isoenzymes play an important role in cellular energy transduction. Two isoenzymes of creatine kinase, ubiquitous mitochondrial creatine kinase (uMtCK) and cytosolic brain creatine kinase (BCK), are postulated to form the creatine phosphate (CP) shuttle, in which creatine serves to transport high-energy phosphate from the mitochondria to its site of utilization. Coordinate regulation of these genes is essential for efficient energy transduction. We examined human CK isoenzyme regulation in placentas during all three trimesters of gestation to define the mRNA and protein expression patterns of uMtCK and BCK and to test the CP shuttle hypothesis. METHODS: Placental samples were collected from a total of 26 patients from the first, second, and third trimesters. Total RNA and protein were prepared from each sample and quantified. Quantitative RNA analysis was performed by gel electrophoresis and dot blot techniques using isoenzyme-specific human cDNA probes for uMtCK and BCK. Protein expression of uMtCK and BCK was examined by Western blot analysis using isoenzyme-specific antibodies to uMtCK and BCK. RESULTS: Analysis of RNA demonstrated the coordinate expression of uMtCK and BCK mRNAs in human placenta, with peak expression of both in the term placentas. Western blot analysis demonstrated coordinate expression of uMtCK and BCK proteins in the first and second trimesters, but not in the term placenta. Expression levels of uMtCK and BCK proteins were not consistent with their respective mRNA levels in the term placenta. CONCLUSION: Expression of uMtCK and BCK in human placenta is highly regulated, and post-transcriptional regulation of uMtCK and BCK expression occurs in the term placenta. The coordinate regulation of uMtCK and BCK in human placenta supports the CP shuttle hypothesis. This analysis demonstrates that human placenta has high energy needs that can change rapidly; thus, a functioning CP shuttle may be important in the maintenance and termination of pregnancy.
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