Skip to main content
SpringerLink
Log in

Regulation of Creatine Kinase Isoenzymes in Human Placenta During Early, Mid-, and Late Gestation

  • Original Article
  • Published:
The Journal of the Society for Gynecologic Investigation: JSGI Aims and scope Submit manuscript

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 UNA 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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bessman SP, Geiger PJ. Transport of energy in muscle: The phosphorylcreatine shuttle. Science 1981;211:448–52.

    Article  CAS  Google Scholar 

  2. Bessman SP, Carpenter CL. The creatine-creatine phosphate energy shuttle. Annu Rev Biochem 1985;54:831–62.

    Article  CAS  Google Scholar 

  3. Stallings RL, Olson E, Strauss AW, Thompson LH, Bachinski LL, Siciliano MJ. Human creatine kinase genes on chromosomes 15 and 19, and proximity of the gene for the muscle form to the genes for apolipoprotein C2 and excision repair. Am J Hum Genet 1988;43:144–51.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Billadello JJ, Kelly OP, Roman DG, Strauss AW. The complete nucleotide sequence of brain B creatine kinase mRNA: Homology in the coding and 3’ noncoding regions among species. Biochem Biophysics Commun 1986;138:392–8.

    Article  CAS  Google Scholar 

  5. Trask RV, Billadello JJ. Tissue-specific distribution and developmental regulation of M and B creatine kinase mRNAs. Biochim Biophys Acta 1990;1049:182–8.

    Article  CAS  Google Scholar 

  6. Friedman DL, Perryman MB. Compartmentation of multiple forms of creatine kinase in the distal nephron of the rat kidney. J Biol Chem 1991;266:22404–10.

    CAS  PubMed  Google Scholar 

  7. Wallimann T, Eppenberger HM. Localization and function of M-line-bound creatine kinase. M-band model and creatine phosphate shuttle. Cell Muscle Motil 1985;6:239–85.

    Article  CAS  Google Scholar 

  8. Saks VA, Rosenshtraukh LV, Smirnov VN, Chazof EI. The role of creatine phosphokinase in cellular function and metabolism. Can J Physiol Pharmacol 1978;56:691–706.

    Article  CAS  Google Scholar 

  9. Wallimann T, Walzthony D, Wegmann G, Moser H, Eppenberger HM, Barrantes FJ. Creatine kinase isoenzymes in spermatozoa. J Muscle Res Cell Motil 1986;7:25–34.

    Article  CAS  Google Scholar 

  10. Schlegel J, Wyss M, Schurch U, et al. Mitochondrial creatine kinase from cardiac muscle and brain are two distinct isoenzymes but both form octameric molecules. J Biol Chem 1991;263:16963–9.

    Google Scholar 

  11. Haas RC, Strauss AW. Separate nuclear genes encode sarcomere-specific and ubiquitous human mitochondrial creatine kinase isoenzymes. J Biol Chem 1990;265:6921–7.

    CAS  PubMed  Google Scholar 

  12. Haas RC, Korenfeld C, Zhang Z, Perryman B, Roman D, Strauss AW. Isolation and characterization of the gene and cDNA encoding human mitochondrial creatine kinase. J Biol Chem 1989;264:2890–7.

    CAS  PubMed  Google Scholar 

  13. Klein SC, Haas RC, Perryman MB, Billadello JJ, Strauss AW. Regulators’ element analysis and structural characterization of the human sarcomeric mitochondrial creatine kinase gene. J Biol Chem 1991;266:18058–65.

    CAS  PubMed  Google Scholar 

  14. Schnyder T, Engel A, Lustig A, Wallimann T. Native mitochondrial creatine kinase forms octameric structures. II. Characterization of dimers and octamers by ultracentrifugation, direct mass measurements by scanning transmission electron microscopy, and image analysis of single mitochondrial creatine kinase octamers. J Biol chem 1988;263:16954–62.

    CAS  PubMed  Google Scholar 

  15. Jacobus WE, Saks VA. Creatine kinase of heart mitochondria: Changes in its kinetic properties induced by coupling to oxidative phosphorylation. Arch Biochem Biophys 1982;219:167–78.

    Article  CAS  Google Scholar 

  16. Payne RM, Friedman DL, Grant JW, Perryman MB, Strauss AW. Creatine kinase isoenzymes are highly regulated during pregnancy in rat uterus and placenta. Am J Physiol 1993;265:E624–35.

    CAS  PubMed  Google Scholar 

  17. Lindahl B, Ahlgren M. Identification of chorion villi in abortion specimens. Obstet Gynecol 1986;67:79–81.

    CAS  PubMed  Google Scholar 

  18. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987;162:156–9.

    Article  CAS  Google Scholar 

  19. Davis LG, Dibner MD, Batter JG. Basic methods in molecular biology. 1st ed. New York: Elsevier, 1986:143–6.

    Book  Google Scholar 

  20. Dichek D, Quertermous T. Thrombin regulation of mRNA levels of tissue plasminogen activator and plasminogen activator inhibitor-1 in cultured human umbilical vein endothelial cells. Blood 1989;74:222–8.

    Article  CAS  Google Scholar 

  21. Payne RM, Haas RC, Strauss AW. Structural characterization and tissue-specific expression of the mRNAs encoding isoenzymes from two rat mitochondrial creatine kinase genes. Biochim Biophys Acta 1991;1089:352–61.

    Article  CAS  Google Scholar 

  22. Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1983;132:6–13.

    Article  CAS  Google Scholar 

  23. Clark JF, Kuznetsov AV, Khuchua Z, Veksler V, Ventura-Clapier R, Saks V. Creatine kinase function in mitochondria isolated from gravid and non-gravid guinea-pig uteri. FEBS Lett 1994;347:147–51.

    Article  CAS  Google Scholar 

  24. Ch’ng JLC, Shoemaker DL, Schimmel P, Holmes EW. Reversal of creatine kinase translational repression by 3’ untranslated sequences. Science 1990;248:1003–6.

    Article  Google Scholar 

  25. Ch’ng JL, Ibrahim B. Transcriptional and posttranscriptional mechanisms modulate creatine kinase expression during differentiation of osteoblastic cells. J Biol Chem 1994;269:2336–41.

    PubMed  Google Scholar 

  26. Jackson RJ. Cytoplasmic regulation of mRNA function: The importance of the 3’ untranslated region. Cell 1993;74:9–14.

    Article  CAS  Google Scholar 

  27. Preiss T, Lightowlers RN. Post-transcriptional regulation of tissue-specific isoforms. J Biol Chem 1993;268:10659–67.

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departments of Obstetrics and Gynecology and Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USA

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

Authors
  1. Michael F. Thomure MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael J. Gast MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Neelam Srivastava
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R. Mark Payne MD
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported in part by a Clinician Scientist Award from the American Heart Association (RMP).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Thomure, M.F., Gast, M.J., Srivastava, N. et al. Regulation of Creatine Kinase Isoenzymes in Human Placenta During Early, Mid-, and Late Gestation. Reprod. Sci. 3, 322–327 (1996). https://doi.org/10.1016/S1071-5576(96)00043-3

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1016/S1071-5576(96)00043-3

Key words

Search

Navigation