Localization and temporal regulation of tissue inhibitors of metalloproteinases 3 and 4 in bovine preovulatory follicles

Abstract

The matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are potential regulators of the focalized extracellular matrix degradation required for ovulation. The objectives of the present study were to determine localization and temporal regulation of TIMP-3 and TIMP-4 mRNA and protein in bovine preovulatory follicles. Ovaries containing preovulatory follicles were collected at 0, 12 and 20 h after GnRH injection for real-time PCR quantification of TIMP-3 and TIMP-4 mRNAs and immunohistochemical localization studies. Additional samples collected at 0, 6, 12, 18 and 24 h post GnRH injection were subjected to Western analysis to determine temporal changes in TIMP-3 and TIMP-4 proteins in the apex and base of preovulatory follicles. Results indicate the gonadotropin surge regulates TIMP-3 and TIMP-4 expression. TIMP-3 and TIMP-4 mRNAs increased within 12 h after GnRH injection. TIMP-3 protein was localized to granulosal and thecal layers of preovulatory follicles and adjacent ovarian stroma, whereas TIMP-4 immunoreactivity was localized to granulosal and thecal cells and ovarian blood vessels. Amounts of TIMP-3 and TIMP-4 proteins in the follicular apex peaked within 12 h post GnRH injection and subsequently declined by 24 h. However, amounts of TIMP-3 and TIMP-4 proteins in the base were not elevated after GnRH administration. Results demonstrate that mRNA and protein for both TIMP-3 and TIMP-4 are increased in bovine preovulatory follicles following the gonadotropin surge. Coordinate expression of TIMPs and MMPs may help regulate the extracellular matrix remodeling characteristic of the ovulatory process.

Ovarian Expression, Localization, and Function of Tissue Inhibitor of Metalloproteinase 3 (TIMP3) During the Periovulatory Period of the Human Menstrual Cycle1

“…In this previous study, staining intensity was highest in the theca interna and luteal cells. An elevated thecal expression of TIMP3 is also seen in other species, such as porcine [47], equine [48], and bovine [42]. This temporal progression of TIMP3 expression in the follicle may allow for the protection of the different cellular layers of the follicle from degradation by matrix metalloproteinases during specific phases of growth and maturation while ensuring the viability of those cells for luteinization as the corpus luteum forms.…”

“…increases approximately 2-fold at 12 h after GnRH injection [42]. Furthermore, it has been shown that TIMP3 mRNA is more abundant in healthy follicles [31,43]; thus, this rise in TIMP3 in the human, bovine, and rat may be required for the final stages of follicular maturation in preparation for the matrix remodeling that takes place before ovulation and expulsion of the oocyte.…”

Cocaine- and Amphetamine-Regulated Transcript Regulation of Follicle-Stimulating Hormone Signal Transduction in Bovine Granulosa Cells

“…RNA isolation, RT, and quantitative real-time RT-PCR analysis were performed as described previously (40,41) using published primer sequences for aromatase and ␤-actin (41,42) and absolute quantification using standard curve technology. Standard curves for each gene were constructed using 10-fold serial dilutions of corresponding plasmids run on the same plates as samples.…”

Expression of tissue inhibitor of metalloproteinase-4 (TIMP-4) in endometrium and placenta of rhesus monkey (Macaca mulatta) during early pregnancy

“…TIMP-4 is a secreted protein expressed at low levels in the placenta (Greene et al, 1996;Leco et al, 1997). In recent years, to conform the role of TIMP-4, many studies were carried out to study its expression, results demonstrated that TIMP-4 was present in several malignant tumors and tumor cell lines (Tunuguntla et al, 2003;Zhao et al, 2004;Jiang et al, 2001;Zhang et al, 2002), as well as in the animal follicle and ovary (Riley et al, 2001;Li et al, 2004;Simpson et al, 2003). It also has been shown that TIMP-4 was expressed in human placenta, fetal membranes, decidua (Greene et al, 1996;Riley et al, 1999), and endometrium Pilka et al, 2004).…”