title: Advantages of pulsatile hormone treatment for assessing hormone-induced gene expression by cultured rat Sertoli cells.
abstract: In response to various hormonal (follicle-stimulating hormone [FSH] and testosterone [T]) and biochemical inputs, testicular Sertoli cells (Sc) produce factors that regulate spermatogenesis. A number of FSH- and T-responsive Sc-specific genes, necessary for spermatogenesis, have been identified to date. However, the hormone-induced in vitro expression pattern of most of these genes is reported to be inconsistent at various time points in primary rat Sc cultures. As a matter of convenience, cultured Sc are constantly exposed to hormones for a few hours to days in the reported literature, although Sc are exposed to pulsatile FSH and T in vivo. The major aim of the present study is to evaluate the advantage, if any, of the in vitro administration of pulsatile hormone (FSH and T in combination) treatment on gene expression of cultured Sc as compared with that of constant hormone treatment. Pulsatile treatment (a 30-min hormonal exposure every 3 h) mimicking the in vivo condition reveals a more prominent effect of hormones in augmenting gene expression as compared with constant treatment. Our results indicate that the expressions of Stem cell factor (Scf, only responsive to FSH), Claudin11 (only responsive to T) and Transferrin (both FSH- and T-responsive) mRNAs are significantly higher at 12 h upon pulsatile treatment than upon constant hormonal treatment. Maximal expression of relevant genes because of pulsatile treatment with hormones suggests that this protocol provides a more suitable premise for assessing hormone-induced gene expression in isolated Sc than one involving constant exposure to hormones.
authors.affiliation: Cellular Endocrinology Laboratory, National Institute of Immunology, ArunaAsaf Ali Marg, New Delhi, 110067, India.|Department of Zoology, Hemvati Nandan Bahuguna Garhwal University, Srinagar, Uttarakhand, India.
authors.forename: Indrashis
authors.initials: I
authors.lastname: Bhattacharya
authors.affiliation: Cellular Endocrinology Laboratory, National Institute of Immunology, ArunaAsaf Ali Marg, New Delhi, 110067, India.|The Ken & Ruth Davee Department of Neurology, Northwestern University, Chicago, Ill., USA.
authors.forename: Mukesh
authors.initials: M
authors.lastname: Gautam
authors.affiliation: Cellular Endocrinology Laboratory, National Institute of Immunology, ArunaAsaf Ali Marg, New Delhi, 110067, India.
authors.forename: Hironmoy
authors.initials: H
authors.lastname: Sarkar
authors.affiliation: Cellular Endocrinology Laboratory, National Institute of Immunology, ArunaAsaf Ali Marg, New Delhi, 110067, India.
authors.forename: Mansi
authors.initials: M
authors.lastname: Shukla
authors.affiliation: Cellular Endocrinology Laboratory, National Institute of Immunology, ArunaAsaf Ali Marg, New Delhi, 110067, India. subeer@nii.ac.in.
authors.forename: Subeer S
authors.initials: SS
authors.lastname: Majumdar
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D057167:Claudins / Q000235:genetics / Q000378:metabolism; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D008297:Male; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017208:Rats, Wistar; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism; D011962:Receptors, FSH / Q000235:genetics / Q000378:metabolism; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism*; D019089:Stem Cell Factor / Q000235:genetics / Q000378:metabolism; D013739:Testosterone / Q000494:pharmacology; D014168:Transferrin / Q000235:genetics / Q000378:metabolism
chemical_list: D057167:Claudins; C492353:Cldn11 protein, rat; D012333:RNA, Messenger; D011944:Receptors, Androgen; D011962:Receptors, FSH; D019089:Stem Cell Factor; D014168:Transferrin; D013739:Testosterone; D005640:Follicle Stimulating Hormone
keywords: Follicle-stimulating hormone; Gene expression; Sertoli cells; Spermatogenesis; Testosterone