title: Localisation and endocrine control of hyaluronan synthase (HAS) 2, HAS3 and CD44 expression in sheep granulosa cells.
abstract: The aim of the present study was to investigate the hormonal regulation of hyaluronan (HA) components in sheep granulosa cells. HA components are present in the reproductive tract and have a range of physical and signalling properties related to reproductive function in several species. First, abattoir-derived ovaries of sheep were used to determine the localisation of HA synthase (HAS) 1-3 and CD44 proteins in antral follicles. Staining for HAS1-3 and CD44 proteins was most intense in the granulosa layer. Accordingly, the expression of HAS2, HAS3 and CD44 mRNA was measured in cultured granulosa cells exposed to 0-50ngmL(-1) of 17β-oestradiol and different combinations of oestradiol, gonadotropins, insulin-like growth factor (IGF)-1 and insulin for 48-96h (1ngmL(-1) FSH, 10ngmL(-1) insulin, 10ngmL(-1) IGF-1, 40ngmL(-1) E2 and 25ngmL(-1) LH.). mRNA expression was quantified by real-time polymerase chain reaction using a fold induction method. The results revealed that the hormones tested generally stimulated mRNA expression of the genes of interest in cultured granulosa cells. Specifically, oestradiol, when combined with IGF-1, insulin and FSH, stimulated HAS2 mRNA expression. Oestradiol and LH had synergistic effects in increasing HAS3 mRNA expression. In conclusion, we suggest that the hormones studied differentially regulate HAS2, HAS3 and CD44 in ovine granulosa cells in vitro. Further work is needed to address the signalling pathways involved.
authors.affiliation: Reproduction Research Group, Royal Veterinary College, Hawkshead Campus, Hawkshead Lane, Hatfield, AL9 7TA, UK.
authors.forename: R
authors.initials: R
authors.lastname: Chavoshinejad
authors.affiliation: Reproduction Research Group, Royal Veterinary College, Hawkshead Campus, Hawkshead Lane, Hatfield, AL9 7TA, UK.
authors.forename: W F A
authors.initials: WF
authors.lastname: Marei
authors.affiliation: Warwick Medical School, University of Warwick, Coventry, CV2 2DX, UK.
authors.forename: G M
authors.initials: GM
authors.lastname: Hartshorne
authors.affiliation: Reproduction Research Group, Royal Veterinary College, Hawkshead Campus, Hawkshead Lane, Hatfield, AL9 7TA, UK.
authors.forename: A A
authors.initials: AA
authors.lastname: Fouladi-Nashta
mesh_terms: D000003:Abattoirs; D000818:Animals; D002478:Cells, Cultured; D004790:Enzyme Induction; D004958:Estradiol / Q000819:agonists / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D014453:Glucuronosyltransferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006107:Granulosa Cells / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism; D000076002:Hyaluronan Synthases; D007150:Immunohistochemistry; D007328:Insulin / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D034461:Luteinization; D007986:Luteinizing Hormone / Q000819:agonists / Q000378:metabolism; D010053:Ovary / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D021381:Protein Transport; D012333:RNA, Messenger / Q000378:metabolism; D034561:Sheep, Domestic
chemical_list: D018960:Hyaluronan Receptors; D007328:Insulin; D007527:Isoenzymes; D012333:RNA, Messenger; D004958:Estradiol; D007334:Insulin-Like Growth Factor I; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D014453:Glucuronosyltransferase; D000076002:Hyaluronan Synthases