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Tuck et al. Journal of Ovarian Research (2015) 8:31 DOI 10.1186/s13048-015-0159-xRESEARCHOpen AccessExpression and localisation of c-kit PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25112874 and KITL in the adult human ovaryAstrud R Tuck1,2, Rebecca L Robker1, Robert J Norman1, Wayne D Tilley2 and Theresa E Hickey1,2*AbstractBackground: The c-kit/kit ligand (KITL) signalling axis is an essential component of ovarian folliculogenesis in mammals, but little is known about expression and localisation of its key components in the ovaries of reproductive age women. This study aimed to characterise mRNA expression of c-kit and KITL isoforms and the localisation of c-kit and KITL proteins in adult human premenopausal ovaries. Methods: This study utilised granulosa cells obtained from the preovulatory follicles of women undergoing assisted reproduction, pieces of ovarian tissue obtained from premenopausal women undergoing gynaecological surgeries and archival paraffin-embedded premenopausal ovarian tissues. Methodology included PCR for gene expression and Western blot or immunohistochemistry for protein expression. Results: Both c-kit mRNA isoforms, known as GNNK+ and GNNK-, were detected in human ovarian cortex, while KITL protein isoforms (KITL1 and KITL2) were present in ovarian cortex and human granulosa cells. Immunohistochemistry showed expression of KITL and c-kit protein in multiple cell types within follicles throughout development, from primordial follicles to large antral follicles, in addition to atretic follicles. Oocytes of all follicle stages expressed c-kit protein exclusively. Interestingly, unlike animal models, expression of both proteins displayed a less cell-type specific distribution with immunostaining present in granulosa, theca and stromal cells, suggesting that autocrine signalling occurs within the human ovary. Conclusion: The results of this study indicate that c-kit/KITL signalling also occurs in the human ovary, as established in various animal models, and may involve previously unknown autocrine signalling. Keywords: Human, Ovarian follicles, c-kit, KITL, Immunohistochemistry, PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28667899 Granulosa, Theca, PCOSBackground KITL is a cytokine growth factor secreted by granulosa cells of ovarian follicles that exerts intra-follicular paracrine signals via stimulation of the c-kit receptor, which is expressed in theca cells and oocytes [1?]. Transgenic mouse models have been utilised to demonstrate critical roles for the key ligand and receptor components of the c-kit/KITL signalling pathway during folliculogenesis ([4?]; 2004). Expression and function of these key components have also been elucidated in other animal models including rat, rabbit, chicken, goat, pig and cow [1, 2, 9?1], and these have demonstrated several key roles that are essential for the maintenance* Correspondence: [email protected] 1 Robinson Research SCR7MedChemExpress SCR7 Institute, School of Paediatrics and Reproductive Health, Adelaide, South Australia, Australia 2 Dame Roma Mitchell Cancer Research Laboratories, School of Medicine; University of Adelaide, Adelaide, South Australia, Australiaof ovarian function and fertility. These roles include primordial follicle activation, promotion of primary to secondary follicle transition, preantral follicle growth, formation of the theca layer, thecal steroidogenesis and anti-apoptosis [3, 9]. Few studies have examined the expression and function of.
