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M D Griswold

Publications and source records attributed to M D Griswold.

At least 109 records · Page 6Linked to original sources
PubMed1978

Metabolic responses of Sertoli cells in culture to various concentrations of follicle stimulating hormone and cholera toxin.

The concentration of cholera toxin required for half-maximal stimulation of cAMP production by Sertoli cell enriched cultures (4.48 X 10(2) microgram/ml) is greater than that required for half-maximal stimulation of 17beta-estradiol synthesis from testosterone (2.34 X 10(-4) microgram/ml), [3H]thymidine incorporation into DNA (1.48 X 10(-5) microgram/ml), or androgen binding protein production (2.43 X 10(-6) microgram/ml). The same relative dose response hierarchy was obtained with respect to stimulation of Sertoli cells with follicle stimulating hormone (FSH) preparations. Again, highest concentrations were required to elicit maximal cAMP production. The data are discussed in relation to an apparent paradox: If cAMP is the mediating 'second messenger' following stimulation by FSH or cholera toxin, why should highest concentrations of these agents be required to elicit 50% of maximal cAMP levels?

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PubMed1978

Paradox of the dose response to polypeptide hormones.

Highest concentrations of follicle-stimulating hormone (FSH) are required to elicit maximal increases in cAMP formation by isolated Sertoli cells in culture, and progressively lower concentrations are required to increase three late responses maximally (estradiol synthesis from testosterone, DNA synthesis, and synthesis of androgen-binding protein, respectively). Identical dose-response curve relations are observed when cholera toxin is used as the stimulatory agent. Results do not rule out the possibility that cAMP could mediate the various responses of Sertoli cells to FSH.

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PubMed1976

FSH stimulation of DNA synthesis in Sertoli cells in culture.

The incorporation of [2H]thymidine into nuclear DNA was investigated in cultured Sertoli cells prepared from testes of 20-day-old rats. Addition of follicle-stimulating hormone (FSH) or dibutyryl cyclic, 3',5'-adenosine monophosphate (DBCAMP) to the culture medium greatly increased incorporation, expressed either as total amounts of [3H]thymidine incorporated per mug DNA or as the percentage of Sertoli cells with labeled nuclear DNA. No stimulation was observed in cells cultured in the presence of testosterone, insulin or cyclic 3',5'-GMP (cGMP). Light and electron microscopic autoradiographic analysis was employed to establish the identity of Sertoli cells having labeled nuclear DNA. Contaminating spermatogonia, which also took up labeled [3H]thymidine, were excluded from cell counts. In addition, Sertoli cells prepared from testes of irradiated 20-day-old germinal cell depleted rats were also observed to incorporate more [3H]thymidine into nuclear DNA when cultured in a chemically defined medium in the presence of FSH. DNA synthesis was abolished by prior treatment of cells with cytosine arabinoside. In separate experiments, the incorporation of [3H]thymidine into DNA of peritubular myoid cells was shown to be independent of FSH or dbcAMP.

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PubMed1972

Temperature-dependent intracellular distribution of thyroxine in amphibian liver.

Rana catesbeiana tadpoles were injected with [(14)C]thyroxine, and the subcellular distribution of the labeled hormone was determined. At 25 degrees the amount of isotope found in the liver was maximal after 1-2 hr and then rapidly decreased to a relatively constant value. A large percentage of the hormone was found associated with the purified nuclei isolated 24 hr after injection of [(14)C]thyroxine. Injection of [(14)C]thyroxine into tadpoles maintained at 5 degrees resulted in a much slower but constant accumulation of isotope in the liver, with virtually no movement of thyroxine into the cell nucleus. Thyroxine was bound very tightly to the chromatin fraction of the nucleus, but extraction and chromatography revealed no chemical modification of the thyroxine itself. These results suggest the presence of two temperature-dependent processes: one concerned with the transport of thyroxine into the liver cell and a second concerned with the transport of the intracellular thyroxine into the cell nucleus. It is proposed that the latter process is involved in the low-temperature inhibition of thyroxine-induced metamorphosis.

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PubMedDate not supplied

Similarity of responses of cultured Sertoli cells to cholera toxin and FSH.

The effects of cholera toxin on the responses of cultured Sertoli cells were compared with those elicited by follicle-stimulating hormone (FSH), and N6O2'-dibutyryl-3',5'-cyclic AMP (bu2cAMP). Addition of FSH or cholera toxin increased cAMP levels. Subsequently there was greater rates of conversion of testosterone to 17beta-estradiol, formation of androgen-binding protein (ABP), and incorporation of [3H]thymidine into DNA by Sertoli cells prepared from testes of immature rats and cultured in the presence of either FSH or cholera toxin. Addition of bu2-cAMP also resulted in enhanced rates of formation of ABP, synthesis of 17beta-estradiol and synthesis of DNA. Cholera toxin and bu2-cAMP elicited changes in morphology of cultured Sertoli cells indistinguishable from those following FSH addition. It is concluded that elevated intracellular cAMP levels can duplicate known actions of FSH on cultured Sertoli cells, but the possible obligatory role of cAMP in mediating FSH actions remains to be evaluated.

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PubMedDate not supplied

Germ cell transplantation and the study of testicular function.

Spermatogonial stem cell transplantation is a novel technique in which donor testicular cells are transferred into recipient testes. A population of germ cells from a transgenic or mutant donor is introduced into the seminiferous tubules of recipient testes by microinjection. Following injections, spermatogonial stem cells can colonize the recipient testis, initiate spermatogenesis and produce sperm capable of fertilization. This technique will allow scientists to: (1) investigate fundamental aspects of spermatogenesis; (2) provide a method to regenerate spermatogenesis in infertile individuals; and (3) genetically manipulate spermatogonial stem cells to develop transgenic animals.

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PubMedDate not supplied

A fragment of prosaposin (SGP-1) from rooster sperm promotes sperm-egg binding and improves fertility in chickens.

A protein isolated from the supernatant of cryopreserved rooster sperm was found to increase the capability of cryopreserved rooster sperm to bind in vitro to the perivitelline membrane of a chicken egg and substantially raise fertility after artificial insemination (AI). That activity was partially purified and termed universal primary sperm-egg binding protein (UPSEBP). Insufficient protein remained from 6 x 10(11) sperm, despite retention of bioactivity, to allow sequencing. We deduced that the protein may be related to prosaposin (also termed SGP-1, for sulfated glycoprotein-1), and we used published amino acid sequences of prosaposin as a guide for synthesis of peptides. Certain peptides were found to increase in vitro sperm-egg binding and increase fertility of frozen-thawed or fresh rooster sperm, in a manner similar to semipurified UPSEBP. Active epitopes were in a 60 amino acid sequence, reflecting the intervening sequence between saposins A and B, plus short extensions into saposins A and B. Highest activity was found when this synthetic peptide was oxidized to form a disulfide bond between terminal cysteines. Antibody against a synthetic peptide consisting of 58 of these 60 amino acids bound to a 7-9 kilodalton protein in UPSEBP. Collectively, the data support the conclusion that UPSEBP is a fragment of prosaposin. Because prosaposin is in semen in humans and animals, these observations have broad implications for possible cause and therapy of one type of subfertility.

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PubMedDate not supplied

E2F and GATA-1 are required for the Sertoli cell-specific promoter activity of the follicle-stimulating hormone receptor gene.

The follicle-stimulating hormone receptor (FSHR) gene is expressed in Sertoli cells in males and in granulosa cells in females. Cis-acting sequences and associated binding factors responsible for the transcription of the TATA-less FSHR gene in Sertoli cells were analyzed with dimethylsulfate (DMS) footprinting assays and electrophoretic mobility shift assay (EMSA). In vivo footprints in the core promoter using nuclear proteins from Sertoli cells identified several protected sequences, including an inverted GATA (TATC, -88/-85), and an E2F (TTTCGCG, -45/-39) motif. EMSA showed the presence of one or more sequence-specific proteins interacting with these potential regulatory elements. Antibody-supershift assays as well as competition assays further revealed that testis-specific GATA-1 recognized the inverted GATA element. The functional role of the potential cis-acting elements was analyzed by transient transfection assays with and without mutations of the putative elements. The mutational analysis indicated that the GATA and E2F elements were each required for optimal promoter activity. The effects of each of the promoter elements was examined in transfections in which mutations were made in each of the known regulatory sites, including the E box, GATA, and E2F sites in various combinations. All of these sites contribute to the maximum promoter activity such that mutations of the E box, GATA, and E2F sites eliminated nearly all promoter activity.

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PubMedDate not supplied

The testicular iron shuttle: a "nurse" function of the Sertoli cells.

The techniques of cell culture and molecular biology first revealed that Sertoli cells synthesize transferrin. Accumulated biological information led to a plausible model for the role of testicular transferrin in an iron shuttle system designed to transport ferric ions around the cellular tight junctions to the germ cells inside the blood-testis barrier. Experiments done in culture and in vivo have supported many aspects of this model. A mutant mouse model that lacks the ability to synthesize transferrin is defective in spermatogenesis and may help to delineate the nature of the iron requirement by germ cells. The levels of seminal transferrin, possibly of Sertoli cell origin, are proportional to sperm production in humans and cattle and may be an effective indicator of Sertoli cell function. The testicular iron shuttle thus represents an important "nurse cell" function of the Sertoli cells.

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