Blocking L-type Voltage-Gated Calcium Ion Channels Changes the Intensity of Protein Synthesis in Metanephric Cells

Yelena Yu. Shapovalova, Liliana A. Kutuzova, Svetlana V. Kharchenko, Svetlana A. Vasilenko

International Journal of Biomedicine. 2019;9(2):150-154.
DOI: 10.21103/Article9(2)_OA14
Originally published June 15, 2019  


The purpose of this research was to study the effect of L-type voltage-gated calcium channels (L-VGCC) blocking on mRNA content and intensity of protein synthesis in cells of the metanephros.
Materials and Methods: The study was performed on 27 outbred Wistar rats weighing 250-270 g from 5 to 7 months of age. Pregnant females were divided into 3 groups of 9 individuals each. Group1 (the control group) included intact pregnant females treated with distilled water; Group 2 (the first experimental group) included animals receiving a therapeutic dose of nifedipine; Group 3 (the second experimental group) included animals receiving a toxic dose of nifedipine. The dihydropyridine nifedipine (Sigma-Aldrich, Gillingham, UK) was used as a Ca2+ channel blocker. Nifedipine was dissolved in distilled water. The dose of nifedipine was calculated based on therapeutic and toxic doses for humans. The therapeutic dose for rats was 127 mg/kg, toxic dose – 762 mg/kg. In each group, when rats reached Day 14, 15, 16 and consistently up to 22 of gestation, they were removed from the experiment by decapitation under ether anesthesia. The embryos and fetuses were removed, subjected to external examination and quickly fixed with 10% neutral formalin entirely up to the age of 15 days. At an older age, both metanephros were removed from the fetuses by preparation and fixed with 10% neutral formalin. The intensity of protein synthesis was estimated by the content of mRNA in the cytoplasm.
Results: The development of the metanephros in rats of the control group was accompanied by a gradual decrease in the content of mRNA, indicating the activity of protein synthesis in the cell cytoplasm of epithelial and mesenchymal buds of the metanephros. In the fetuses of rats whose mothers received therapeutic and toxic doses of nifedipine, statistically significant changes in the content of mRNA in the cytoplasm of the metanephric cells were observed. These changes were most significant in the metanephric cells after administration of the toxic dose. On ED 17, an L-VGCC blockade with a therapeutic dose of nifedipine led to a paradoxical reaction of cells with an increase in protein synthesis, compared with the control group.
Conclusion: Both doses of nifedipine have a greater effect on the content of mRNA in the cytoplasm of epithelial cells, compared to mesenchymal buds.

metanephros • voltage-gated calcium channels • nifedipine • protein synthesis
  1. Kramer I. Intracellular Calcium. In: Signal Transduction. Third Edition. Elsevier; Amsterdam, The Netherlands; 2016:381–439.
  2. Catterall WA. Voltage-gated calcium channels. Cold Spring Harb Perspect Biol. 2011;3(8):a003947. doi: 10.1101/cshperspect.a003947.
  3. Finney BA, del Moral PM, Wilkinson WJ, Cayzac S, Cole M, et al. Regulation of mouse lung development by the extracellular calcium-sensing receptor, CaR. J Physiol.  2008;586:6007–6019. 
  4. Tena JJ, Neto A, de la Calle-Mustienes E, Bras-Pereira C, Casares F, Gómez-Skarmeta JL. Odd-Skipped Genes Encode Repressors That Control Kidney Development. Dev. Biol. 2007;301:518–531. doi: 10.1016/j.ydbio.2006.08.063.
  5.  Compendium 2006 - Medicinal Products. In Kovalenko VN, Viktorov AP, editors. K.: Morion; 2006. [In Russian].
  6. Spirin AS. Biosynthesis of proteins, the world of RNA and the origin of life. Herald of the Russian Academy of Sciences. 2001;71(4):320-328. [Article in Russian].
  7. Kharchenko SV. Changes in the activity of protein synthesis in embryogenesis of the lungs and kidneys of rats after the administration of paracetamol and nimesulide to pregnant females. World of Medicine and Morphology. 2009;5(3-2):185-188. [Article in Russian].
  8. Semchenko VV, Barashkova SA, Nozdrin VI. Histological technique. Omsk.: 2006; 289 p. [In Russian].
  9. Leclerc C, Webb SE, Miller A, Moreau M. An increase in intracellular Ca 2+ is involved in the differentiation of prenodic tubules in amphibians of Xenopus laevis. Dev Biol. 2008;321(2):357-67. doi: 10.1016/j.ydbio.2008.06.029.
  10. Paudel S,  Sindelar R,  Sahа. Calcium Signaling in Vertebrate Development and Its Role in Disease. Int J Mol Sci. 2018;19(11):3390 doi: 10.3390/ijms19113390
  11. Gilbert T, Leclerc C, Moreau M. Control of kidney development by calcium ions. Biochimie. 2011;93(12):2126-31. doi: 10.1016/j.biochi.2011.07.007.
  12. Hughes AD. Calcium channels in vascular smooth muscle cells. J Vasc Res.  1995;32:353–370.
  13. Ghosh D, Syed AU, Prada MP, Nystoriak MA, Santana LF, Nieves-Cintrón M, Navedo MF. Calcium Channels in Vascular Smooth Muscle. Adv Pharmacol. 2017;78:49-87. doi: 10.1016/bs.apha.2016.08.002.
  14. Lin AH, Sun H, Paudel O, Lin MJ, Sham JS. Conformation of ryanodine receptor-2 gates store-operated calcium entry in rat pulmonary arterial myocytes. Cardiovasc Res. 2016;111(1):94-104. doi: 10.1093/cvr/cvw067.
  15. Mukhina AM, Altaeva EG, Nemirovskaya TL, Shenkman BS. The role of L-type calcium channels in the accumulation of Ca2+ in soleus muscle fibers in the rat and changes in the ratio of myosin and serca isoforms in conditions of gravitational unloading. Neurosci Behav Physiol. 2008 Feb;38(2):181-8. doi: 10.1007/s11055-008-0027-x.
  16. Xu H, Sun M, Zhao X. Turing mechanism underlying a branching model for lung morphogenesis. PLoS One. 2017;12(4):e0174946. doi: 10.1371/journal.pone.0174946.
  17. Fontana JM, Khodus GR, Unnersjö-Jess D, Blom H, Aperia A, Brismar H. Spontaneous calcium activity in metanephric mesenchymal cells regulates branching morphogenesis in the embryonic kidney. FASEB J. 2019;33(3):4089-4096. doi: 10.1096/fj.201802054R.
  18. Blake J, Rosenblum ND. Renal branching morphogenesis: morphogenetic and signaling mechanisms. Semin Cell Dev Biol. 2014;36:2-12. doi: 10.1016/j.semcdb.2014.07.011.
  19. Hannezo E, Scheele CLGJ, Moad M, Drogo N, Heer R, Sampogna RV, van Rheenen J, Simons BD. A Unifying Theory of Branching Morphogenesis. Cell. 2017;171(1):242-255.e27. doi: 10.1016/j.cell.2017.08.026.
  20. Kim JM, Choi S, Lee SW, Park K. Voltage-dependent Ca2+ channels promote branching morphogenesis of salivary glands by patterning differential growth. Sci Rep. 2018;8(1):7566. doi: 10.1038/s41598-018-25957-w.
  21. Vetter MR, Gibley CW. Morphogenesis and histochemistry of the developing mouse kidney. Journal of Morphology. 2005;120(2):135-155.

Download Article
Received May 13, 2019.
Accepted June 13, 2019.
©2019 International Medical Research and Development Corporation.