Expression and Regulation of MMP1, MMP3, and MMP9 in the Chicken Ovary in Response to Gonadotropins, Sex Hormones, and TGFB11

Abstract

Matrix metalloproteinases (MMPs) are a specific class of proteolytic enzymes that play critical roles in follicular development and luteinization in mammals. However, the role of MMPs in avian ovary remains largely unknown. We found that three MMP genes (MMP1, MMP3, and MMP9) were significantly up-regulated in 23-wk-old (laying phase) chicken ovaries compared with 6-wk-old ovaries (prepubertal phase). In reproductively active chicken ovary, MMP1 expression (both mRNA and protein) remained low in prehierarchical and preovulatory follicles but increased in postovulatory follicles (POFs). Both MMP3 and MMP9 expression levels increased during follicular maturation. MMP3 reached maximal expression in the first largest follicle (F1), while MMP9 levels continued to rise in POF1 and POF2 after ovulation. Immunohistochemistry, Western blot analysis, and zymography experiments indicated that MMP1, MMP3, and MMP9 were synthesized and secreted by granulosa cells of different follicles in the chicken ovary. The mRNA expression of MMP1 and MMP3 in the granulosa cells was stimulated by follicle-stimulating hormone, luteinizing hormone, progesterone, and estrogen but not by transforming growth factor beta 1 (TGFB1). However, the mRNA of MMP9 was induced by TGFB1 but not follicle-stimulating hormone, luteinizing hormone, progesterone, or estrogen. Luciferase reporter and mutagenesis analysis indicated the AP1 and NFkappaB elements located in the promoter region from -1700 to -2400 bp were critical for both basal and TGFB1-induced MMP9 transcription. These data provide the first spatial-temporal expression analysis of MMP system in the chicken ovary.

A comprehensive transcriptomic view on the role of SMAD4 gene by RNAi-mediated knockdown in porcine follicular granulosa cells

“…Previously, MMP9 has been shown to be expressed in porcine GCs (Basini et al 2011). In chicken ovary, MMP9 mRNA levels in GCs were discovered significantly higher than in theca cells and its protein expression was found in both the conditioned medium and the GC lysate, showing GCs could be considered as important cellular origin of MMP9 protein (Zhu et al 2014). Here our data was consistent with the study by Costello et al (2009), who found that the expression of MMP9 was decreased in SMAD4 mutant embryonic stem cells and embryoid bodies, indicating that SMAD4 functional loss led to decrease the expression of MMP9.…”

Expression pattern of matrix metalloproteinases changes during folliculogenesis in the cat ovary

“…The amplification curves and melting curves of each PCR product were checked to verify the amplification efficiencies and the targeted amplicon. The CT value of each gene was normalized against the average CT of GAPDH (housekeeping gene) to generate delta CT (ΔCT; Zhu et al., ). Target gene expression levels were determined by the comparative threshold cycle (ΔCT) method (Schmittgen & Livak, ).…”

ID3 mediates the TGF‐β1‐induced suppression of matrix metalloproteinase‐1 in human granulosa cells

“…The MMP family belongs to zinc‐ and calcium‐dependent metalloproteinases and encompasses at least 28 members that can be subdivided into four broad classes: collagenases, gelatinases, stromelysins, and membrane‐type MMPs . Several members of the MMP family (e.g., MMP1, MMP2, MMP7, MMP9, and MMP13) are expressed in mammalian ovarian follicles at different developmental stages, and the expression of these MMPs is highly regulated in response to gonadotropins, sex hormones, and growth factors . For instance, MMP1 and MMP9 are expressed in granulosa cells at all follicular stages, and MMP2 is expressed in the granulosa/cumulus cells of early and antral follicles .…”