Regulation of creatine kinase isoenzymes in human placenta during early, mid-, and late gestation

Michael F. Thomure, Michael J. Gast, Neelam Srivastava, R. Mark Payne

Research output: Contribution to journalArticle

10 Citations (Scopus)

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 cystolic brain creatine kinase (BCK), and 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 trimester 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.

Original languageEnglish (US)
Pages (from-to)322-327
Number of pages6
JournalJournal of the Society for Gynecologic Investigation
Volume3
Issue number6
DOIs
StatePublished - Nov 1 1996

Fingerprint

Mitochondrial Form Creatine Kinase
BB Form Creatine Kinase
Creatine Kinase
Placenta
Isoenzymes
Pregnancy
Phosphocreatine
Second Pregnancy Trimester
RNA
First Pregnancy Trimester
Proteins
Messenger RNA
Western Blotting
Pregnancy Maintenance
Creatine
Essential Genes
Third Pregnancy Trimester
Electrophoresis

Keywords

  • Creatine kinase
  • mitochondria
  • placenta
  • smooth muscle

ASJC Scopus subject areas

  • Obstetrics and Gynecology

Cite this

Regulation of creatine kinase isoenzymes in human placenta during early, mid-, and late gestation. / Thomure, Michael F.; Gast, Michael J.; Srivastava, Neelam; Payne, R. Mark.

In: Journal of the Society for Gynecologic Investigation, Vol. 3, No. 6, 01.11.1996, p. 322-327.

Research output: Contribution to journalArticle

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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 cystolic brain creatine kinase (BCK), and 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 trimester 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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AB - OBJECTIVE: Creatine kinase (CK) isoenzymes play an important role in cellular energy transduction. Two isoenzymes of creatine kinase, ubiquitous mitochondrial creatine kinase (uMtCK) and cystolic brain creatine kinase (BCK), and 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 trimester 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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