title: Characterisation of an in vitro system to study maternal communication with spermatozoa.
abstract: In vivo, gamete maturation, fertilisation and early embryonic development take place inside the oviduct. Several studies have indicated that local responses towards gametes and embryos are generated by the maternal reproductive tract. However, no defined in vitro model currently exists to allow detailed and systematic investigation of maternal communications with gametes and embryos. Therefore, we characterised an in vitro model based on the interaction of boar spermatozoa with an immortalised porcine oviduct epithelial cell line to evaluate different factors that may affect this model. The factors tested were sperm viability, source of spermatozoa, cell passage effect and the effect of reproductive and non-reproductive epithelial cells in the interaction with spermatozoa. After 24 h of co-incubation, RNA was extracted and used to synthesise cDNA for quantitative real-time PCR. Alteration in the expression of genes such as adrenomedullin, heat-shock 70-kDa protein 8 and prostaglandin E synthase was considered as the end point of this assay. The results showed that sperm viability and cell passage number had an effect on oviductal gene expression in response to spermatozoa. Oviductal cells showed significant alterations in gene expression when compared with non-reproductive epithelial cells. The simple in vitro system described here has potential application for further studies in our understanding of mechanisms involved in maternal interactions with spermatozoa.
authors.affiliation: Academic Unit of Reproductive and Developmental Medicine, University of Sheffield, Sheffield S10 2SF, UK.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Aldarmahi
authors.forename: Sarah
authors.initials: S
authors.lastname: Elliott
authors.forename: Jean
authors.initials: J
authors.lastname: Russell
authors.forename: Thomas
authors.initials: T
authors.lastname: Klonisch
authors.forename: Sabine
authors.initials: S
authors.lastname: Hombach-Klonisch
authors.forename: Alireza
authors.initials: A
authors.lastname: Fazeli
mesh_terms: D053607:Adrenomedullin / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002450:Cell Communication / Q000235:genetics; D049109:Cell Proliferation; D002470:Cell Survival; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000378:metabolism / Q000502:physiology*; D005260:Female; D005786:Gene Expression Regulation; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D019746:Intramolecular Oxidoreductases / Q000235:genetics / Q000378:metabolism; D018374:LLC-PK1 Cells; D008297:Male; D010057:Oviducts / Q000166:cytology*; D000071876:Prostaglandin-E Synthases; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013045:Species Specificity; D013094:Spermatozoa / Q000378:metabolism / Q000502:physiology*; D013552:Swine; D013997:Time Factors
chemical_list: D018840:HSP70 Heat-Shock Proteins; D012333:RNA, Messenger; D053607:Adrenomedullin; D019746:Intramolecular Oxidoreductases; D000071876:Prostaglandin-E Synthases