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M Loir

Publications and source records attributed to M Loir.

14 recordsLinked to original sources
PubMed1992

Evidence for binding and action of growth hormone in trout testis.

Growth hormone (GH) binding to testis tissue and GH action on trout testicular cells were studied in vitro. Labeled salmon GH (sGH) was able to bind to a trout testis membrane preparation. Binding sites showed high affinity (Ka = 1-2 x 10(9) M-1) and low capacity (11 fmol/g fresh tissue) for 125I-sGH. Salmon GH and bovine GH, but not salmon gonadotropin, could compete with 125I-sGH for site occupancy. The binding characteristics were similar to those of trout liver GH receptors that we previously described. Salmon GH (0.1 and 1 microgram/ml) and bovine GH (10 micrograms/ml) could modulate steroidogenesis in cultured testicular cells: 17 alpha-hydroxy, 20 beta-dihydroprogesterone (17 alpha 20 beta OHP) accumulation in culture medium was stimulated by GH addition, and this effect increased with duration of culture and/or stimulation; 11-ketotestosterone accumulation tended to be inhibited in the presence of GH at the beginning of culture. These effects were dependent on GH concentration and were observed both in the absence and presence of gonadotropin. The amplitude of the sGH effect varied between experiments, probably according to the physiological state of the cells used. In vivo, GH and 17 alpha 20 beta OHP plasma levels increased at the beginning of spermiation (sperm production) and decreased at the end of spermiation. This relationship suggests that, at the end of the reproductive cycle, high GH levels are associated with the production of 17 alpha 20 beta OHP, a progestin necessary for efficient spawning in this species. We conclude that GH may play a role in testicular physiology, at least at certain stages of spermatogenesis.

Animals↗
PubMed1991

Nuclear transition protein 1 from ram elongating spermatids. Mass spectrometric characterization, primary structure and phosphorylation sites of two variants.

The ram transition protein 1 (TP1) is present in spermatid cell nuclei in the nonphosphorylated, monophosphorylated and diphosphorylated forms. Its primary structure was determined by automated Edman degradation of S-carboxamidomethylated protein and of peptides generated by cleavage with thermolysin and endoproteinase Lys-C. The ram TP1 is a small basic protein of 54 residues and structurally very close to other mammalian TP1. The mass spectrometric data obtained from the protein and its fragments reveal that ram TP1 is indeed a mixture (approximately 5:1) of two structural variants (Mr 6346 and 6300). These variants differ only by the nature of the residue at position 27 (Cys in the major variant and Gly in the minor variant). The study of phosphorylation sites has shown that four different serine residues could be phosphorylated in the monophosphorylated TP1, at positions 8, 35, 36 or 39. From previous physical studies, it has been postulated that the Tyr32 surrounded by two highly conserved basic clusters was responsible for the destabilization of chromatin by intercalation of its phenol ring between the bases of double-stranded DNA. The presence of three phosphorylatable serine residues in the very conserved sequence 29-42 is another argument for the involvement of this region in the interaction with DNA.

Amino Acid Sequence↗
PubMed1990

Trout steroidogenic testicular cells in primary culture. I. Changes in free and conjugated androgen and progestagen secretions: effects of gonadotropin, serum, and lipoproteins.

Isolated trout steroidogenic testicular cells were cultured for 10-15 days, either mixed with other round cells or after enrichment in interstitial cells. Free and conjugated progestagen and androgen secretions were assayed using specific radioimmunoassays (RIA). Free progesterone, 17 alpha-hydroxyprogesterone (17 alpha-OH-P), 17 alpha-hydroxy,20 beta-dihydroprogesterone (17 alpha,20 beta-OH-P), androstenedione, testosterone (T), and 11-ketotestosterone (11KT) were produced by testicular cells prepared from testes in spermatogenesis and mature testes. Discrete amounts of dehydroepiandrosterone (DHA) and of estradiol were secreted by mixed testicular cells prepared from mature testes, but no estradiol was detected in interstitial cell media. Conjugated androgens were produced by interstitial cells. While the production of progestagens by cells from spermatogenetic and mature testes either remained constant or increased throughout culture duration, those of free and conjugated androgens progressively decreased to low values whatever the components added to the medium. When salmon gonadotropin (s-GtH) was present permanently, androgen (free and conjugated) and progestagen secretions were stimulated for 3 to 4 days. When GtH was present discontinuously (1 day in every 3 days), the sensitivity of the cells was maintained for at least 7 days. While the GtH-stimulated/basal ratio was high for androgens, it was rather low for 17 alpha 20 beta-OH-P as compared to the values obtained with testis fragments. Trout serum (5%) stimulated the secretion of free and conjugated T and 11KT when testes were mature, but not when they were in spermatogenesis, while it stimulated 17 alpha 20 beta-OH-P secretion at the two stages. Total trout lipoproteins (125-500 micrograms/ml) stimulated 17 alpha 20 beta-OH-P secretion by cells from spermatogenetic testes, but not 11KT secretion.

17-alpha-Hydroxyprogesterone↗
PubMed1990

Trout steroidogenic testicular cells in primary culture. II. Steroidogenic activity of interstitial cells, Sertoli cells, and spermatozoa.

Somatic cells (interstitial cells and Sertoli cells) were prepared either as single cells or in clusters, from spermatogenic and mature trout testes, according to Loir (1988), and cultured for 10-14 days. Sertoli cells are 3 beta-HSD negative when prepared from testes resuming spermatogenesis and from mature testes, but they are 3 beta-HSD positive in spermatogenic testes. Progesterone, 17 alpha-hydroxyprogesterone (17 alpha-OH-P), and free androgens are secreted by interstitial cells, 11-ketotestosterone (11KT) being the predominating steroid produced immediately after seeding. These cells also produce high levels of glucuronated androgens. At least in mature spermiating testes they do not secrete estradiol. After isolation, interstitial cells would lose most of their ability to secrete 17 alpha-hydroxy,20 beta-dihydroprogesterone (17 alpha 20 beta-OH-P) but they would recover it later. Testicular spermatozoa, which convert 17 alpha-OH-P independently of s-GtH, constitute a second source of this progestagen. In addition, our results suggest that Sertoli cells could be able to secrete 17 alpha-OH-P and also progesterone. A possible participation of the intralobular production of the former progestagen to the local regulation of germ cell maturation is evoked.

17-alpha-Hydroxyprogesterone↗
PubMed1977

Steroid release in vitro by two luteal cell types in the corpus luteum of the pregnant sow.

Corpora lutea from sows at 30, 60 and 90 days of gestation were dissociated enzymically, and the components of the resulting cell suspension were separated by sedimentation at unit gravity. Two luteal cell populations of 30-50 micronm diameter and 15-20 micronm diameter were obtained and superfused for up to 18 h with Dulbecco's modified Eagle medium, the cells being supported in a column in a matrix of Biogel. Fractions were collected every 30 min and assayed for progesterone and oestradiol-17beta. At 30 and 60 days of gestation the large luteal cells produced progesterone at an initial rate of approximately 100 ng/h/10(5) cells, which decreased to half this rate at 90 days. The smaller cells also released progesterone into the medium at approximately 15-20 ng/h/10(5) cells at all stages of gestation. At 30 days of gestation, neither cell type released significant amounts of oestradiol-17beta, but from 60 days onwards ,significant and increasing quantities were measured in the superfusates from the larger cells. Both cell types were perfused with porcine LH at the three stages of gestation, and both showed an immediate response in terms of progesterone release which decreased in magnitude with increasing age of gestation. The response of the smaller cells was greater than that of the larger cells.

Animals↗
PubMed1976

Seasonal variations in the response of the testis and LH levels to hemicastration of adult rams.

The weight and histology of the testis and plasma LH levels were analysed after hemicastration of adult Ile-de-France rams in spring or in autumn. After hemicastration, the remaining testes were significantly heavier than those of entire animals measured at the same time of year. At 4 or 6 months after hemicastration performed in spring, the remaining testes were hypertrophied by nearly 40% as compared to the testes of entire sexually active animals, assessed in autumn. The variations of intertubular tissue volume, total seminiferous tubule length, stem cell stocks, daily production of round spermatids, and cellular volume of primary spermatocytes paralleled the variations in testis weight. The annual decrease of the area of the Sertoli cell nuclei and of the yield of meiosis and beginning of spermiogenesis during the non-breeding season was prevented by hemicastration performed in autumn. Plasma LH levels were consistently elevated till autumn after hemicastration performed in spring. A positive and significant correlation was observed between LH levels and yields of spermatogonial divisions.

Animals↗