Molecular characterization and regulation of the angiotensin-converting enzyme type 2/angiotensin-(1-7)/MAS receptor axis during the ovulation process in cattle

J Renin Angiotensin Aldosterone Syst. 2012 Mar;13(1):91-8. doi: 10.1177/1470320311417273. Epub 2011 Aug 8.

Abstract

The objective of this study was to characterize the profiles of Ang-(1-7), MAS receptor, ACE(2), NEP and PEP during the ovulatory process in cattle. For this study, 40 synchronized cows with follicular diameter ≥ 12 mm were ovariectomized at different time-points (0, 3, 6, 12 and 24 h) after i.m. application of gonadotropin-releasing hormone (GnRH) to induce a luteinizing hormone surge. Follicular fluid was collected for measuring Ang-(1-7) by radioimmunoassay. Theca and granulosa cells were isolated from the preovulatory follicles to evaluate the gene expression of MAS receptor, ACE(2), NEP and PEP by qRT-PCR assay. Cross-contamination between theca and granulosa cells was tested by RT-PCR to detect cytochrome P450 aromatase (CYP19A1) and 17α-hydroxylase (CYP17A1) mRNA. Ang-(1-7) levels were constant until 12 h and then increased (p < 0.05) at 24 h after GnRH. Messenger RNA expression of MAS, ACE(2), NEP and PEP was detected in theca and granulosa cells at all time-points after GnRH. In granulosa cells, ACE(2), NEP and PEP were differentially expressed after GnRH treatment (p < 0.05). In conclusion, the Ang-(1-7), MAS receptor, ACE(2), NEP and PEP profiles in preovulatory follicles indicate that Ang-(1-7) plays a role in the regulation of the ovulatory process in cattle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin I / genetics*
  • Angiotensin I / metabolism
  • Angiotensin-Converting Enzyme 2
  • Animals
  • Cattle
  • Female
  • Follicular Fluid / metabolism
  • Gene Expression Regulation*
  • Granulosa Cells / metabolism
  • Models, Animal
  • Neprilysin / genetics
  • Neprilysin / metabolism
  • Ovariectomy
  • Ovulation / genetics*
  • Peptide Fragments / genetics*
  • Peptide Fragments / metabolism
  • Peptidyl-Dipeptidase A / genetics*
  • Peptidyl-Dipeptidase A / metabolism
  • Prolyl Oligopeptidases
  • Proto-Oncogene Mas
  • Proto-Oncogene Proteins / genetics*
  • Proto-Oncogene Proteins / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Receptors, G-Protein-Coupled / genetics*
  • Receptors, G-Protein-Coupled / metabolism
  • Reproducibility of Results
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / metabolism
  • Signal Transduction / genetics*
  • Theca Cells / metabolism

Substances

  • Peptide Fragments
  • Proto-Oncogene Mas
  • Proto-Oncogene Proteins
  • RNA, Messenger
  • Receptors, G-Protein-Coupled
  • Angiotensin I
  • Peptidyl-Dipeptidase A
  • Angiotensin-Converting Enzyme 2
  • Serine Endopeptidases
  • Prolyl Oligopeptidases
  • Neprilysin
  • angiotensin I (1-7)