scholarly journals Culture patterns and sorting of rat Sertoli cell secretory proteins

1988 ◽  
Vol 89 (2) ◽  
pp. 175-188
Author(s):  
H. Ueda ◽  
L.L. Tres ◽  
A.L. Kierszenbaum
Keyword(s):  
Epithelial Cells ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
Secretory Proteins ◽  
Spermatogenic Cells ◽  
Squamous Cells ◽  
Nylon Mesh ◽  
Continuous Sheet ◽  
Peritubular Cells ◽  
Cells Cultured

A cocultivation chamber and two types of permeable substrates have been used to study: (1) the culture patterns of rat Sertoli and peritubular cells, and Sertoli cells cocultured with spermatogenic cells or peritubular cells; and (2) the polarized secretion of Sertoli cell-specific proteins transferrin, S70 and S45-S35 heterodimeric protein. Substrates included a nylon mesh (with openings of 100 micron) coated with extracellular matrix (ECM) material and an uncoated microporous filter (with pores of 0.45 micron). Sertoli cells cultured on ECM-coated nylon mesh organized a continuous sheet of multilayered epithelial cells essentially devoid of spermatogenic cells while peritubular cells formed a layer of squamous cells. Sertoli cells cultured on uncoated microporous substrate formed a continuous sheet of cuboidal epithelial cells with numerous basal cytoplasmic processes projecting into the substrate and abundant apically located spermatogenic cells, while peritubular cells organized one or two layers of loose squamous cells. [35S]methionine-labelled secretory proteins resolved by two-dimensional polyacrylamide gel electrophoresis and autoradiography displayed cell-specific patterns that were slightly influenced by the type of substrate. Sertoli cells cocultured with peritubular cells on uncoated microporous substrate under conditions that enabled separation of apical and basal surfaces, secreted proteins in a polarized fashion. While transferrin was released bidirectionally, S45-S35 heterodimeric protein was released apically. S70 was detected in both apical and basal compartments. We conclude from these studies that: (1) the number of spermatogenic cells decreases when Sertoli-spermatogenic cell cocultures are prepared on ECM-coated nylon substrate; and (2) Sertoli cells in coculture with spermatogenic or peritubular cells on uncoated microporous substrate, organize continuous sheets displaying polarized protein secretion.

scholarly journals Thyroid hormone effects on androgen receptor messenger RNA expression in rat Sertoli and peritubular cells

1998 ◽  
Vol 156 (1) ◽  
pp. 43-50 ◽  
Author(s):  
NK Arambepola ◽  
D Bunick ◽  
PS Cooke
Keyword(s):  
Androgen Receptor ◽  
Thyroid Hormone ◽  
Mrna Expression ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
Mrna Levels ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Peritubular Cells

Postnatal Sertoli cell maturation is characterized by a pronounced rise in androgen receptor (AR) expression, which increases several fold between birth and adulthood. Since both 3,3',5-triiodothyronine (T3) and FSH regulate Sertoli cell proliferation and differentiation, we have determined the effects of T3 and FSH on AR mRNA expression in cultured Sertoli cells from 5-day-old rats. These cultures contain 5-9% peritubular cells, which also express AR mRNA. To insure that the observed T3 responses did not result from peritubular cells, we examined T3 effects on AR mRNA expression in cultured 20-day-old Sertoli cells (which contain minimal peritubular contamination) and peritubular cells, and measured thyroid hormone receptor (TR) mRNA expression in both of these cell types. Sertoli cells from 5- and 20-day-old rat testes were grown in serum-free medium alone (controls) or with ovine FSH (100 ng/ml) and/or T3 (100 nM) for 4 days. Peritubular cells purified from 20-day-old rat testes were grown in serum-containing medium for 8 days. These cells were split 1:4, and grown an additional 8 days, the last 4 days in serum-free medium with or without T3. TR and AR mRNA levels in all cultures were determined by Northern blotting. AR mRNA levels in 5- and 20-day-old cultured Sertoli cells were significantly (P < 0.05) increased by both T3 and FSH alone. Furthermore, AR mRNA levels in Sertoli cells treated with T3 and FSH were greater than with either alone. TR mRNA expression was detected in cultured peritubular cells, but TR mRNA levels in these cells were only approximately 30% of that seen in 20-day-old cultured Sertoli cells. In contrast to Sertoli cells, T3 did not affect peritubular AR mRNA expression. These results indicate that T3 is an important regulator of the postnatal Sertoli cell AR mRNA increase. The additive effects of maximally stimulatory doses of FSH and T3 suggest these hormones work through different mechanisms to increase AR mRNA. TR mRNA expression in peritubular cells indicates these cells may be direct T3 targets, though the function of T3 in these cells is unknown.

scholarly journals Disappearance of Vimentin in Sertoli Cells: A Mono(2-ethylhexyl) Phthalate Effect

2007 ◽  
Vol 26 (4) ◽  
pp. 289-296 ◽  
Author(s):  
Tat Wei Tay ◽  
Bibin Bintang Andriana ◽  
Maki Ishii ◽  
Naoki Tsunekawa ◽  
Yoshiakira Kanai ◽  
...  
Keyword(s):  
Sertoli Cell ◽  
Cell Cultures ◽  
Sertoli Cells ◽  
Annexin V ◽  
Fluorescein Isothiocyanate ◽  
Blue Staining ◽  
Spermatogenic Cells ◽  
Gradual Disappearance ◽  
Ethylhexyl Phthalate

The effects of mono(2-ethylhexyl) phthalate (MEHP) on 21-day-old C57Bl/6N mice and their Sertoli cell cultures were studied. Mice were given a single dose of 800 mg/kg MEHP by oral gavage and sacrificed 24 h later. At the same time, testes were harvested from another batch of mice for Sertoli cell cultures. Cultures were subsequently exposed to 0, 1, and 100 nmol/ml MEHP for 0, 3, 6, 12, and 24 h. An antivimentin antibody was used to detect intermediate filament changes in Sertoli cells. Meanwhile, detection of preapoptotic signals and presence of apoptotic cells were done using annexin V–FITC (fluorescein isothiocyanate) and TUNEL (deoxynucleotidyltransferase-mediated dUTP nick end labeling) analyses, respectively. In vivo results showed a correlation between the increase in TUNEL-positive cells and the vimentin disruption in treated mice. Toluidine blue staining of the Sertoli cell cultures showed the increased number and size of vacuoles in Sertoli cell cytoplasm. Vimentin immunohistochemistry showed gradual disappearance of vimentin in Sertoli cell cultures as time and dose increased. Some Sertoli cells were found to be annexin V–FITC positive, but no TUNEL-positive cells were found. Taken together, these results show that the appearance of vacuoles and the vimentin disappearance caused by MEHP in the Sertoli cells are related with each other and can be observed in relation to time. This can be used as an indicator of the loss of mechanical support for spermatogenic cells, which in the end causes apoptosis of spermatogenic cells.

scholarly journals A HISTOLOGICAL AND CHEMICAL STUDY OF THE FATTY MATTER OF NORMAL AND CRYPTORCHID TESTES

1911 ◽  
Vol 13 (3) ◽  
pp. 355-364 ◽  
Author(s):  
Frederic M. Hanes ◽  
Jacob Rosenbloom
Keyword(s):  
Sertoli Cell ◽  
Sertoli Cells ◽  
Dry Weight ◽  
Chemical Study ◽  
Excessive Amount ◽  
Fatty Material ◽  
Spermatogenic Cells ◽  
Fatty Matter

1. About 19 per cent. of the dry weight of the normal pig testicle is fatty matter. Histologically this fat occurs largely in the cells of the seminal tubules, and especially in the Sertoli cells. 2. During spermatogenesis the fat of the Sertoli cell passes inward for the nutrition of the spermatids and spermatozoa. During this passage its character is altered from a neutral fat to a lipoid. 3. About 30 per cent. of the dry weight of the cryptorchid pig testicle is fatty material. Histologically this fat lies within the seminal tubules, partially filling the Sertoli cells. The spermatogenic cells have completely disappeared. 4. We conclude that the presence of such an excessive amount of fat in the cryptorchid testicle is due to the absence of the spermatogenic cells which normally utilize during their development the fat furnished by the Sertoli cells.

GONADOTROPHINS, TESTOSTERONE AND SPERMATOGENESIS IN NEONATALLY IRRADIATED MALE RATS: EVIDENCE FOR A ROLE OF THE SERTOLI CELL IN FOLLICLE-STIMULATING HORMONE FEEDBACK

1977 ◽  
Vol 75 (2) ◽  
pp. 209-219 ◽  
Author(s):  
F. H. DE JONG ◽  
R. M. SHARPE
Keyword(s):  
Sertoli Cell ◽  
Sertoli Cells ◽  
Age Groups ◽  
Male Rats ◽  
Male Rat ◽  
Postnatal Life ◽  
Spermatogenic Cells ◽  
Normal Numbers ◽  
Testicular Cell

Peripheral concentrations of FSH in the male rat seem to be regulated in part by a protein hormone, inhibin, which originates from the testes. In an attempt to ascertain which type of testicular cell secretes inhibin, groups of male rats were irradiated prenatally or on days 4, 6 or 8 of postnatal life, and killed at 21, 51 or 81 days of age together with castrated and intact controls. The concentrations of FSH and LH in the pituitary gland, and FSH, LH and testosterone in the plasma were estimated for each animal, and the numbers of each class of intratubular cell in the testes were calculated. Rats irradiated neonatally had fewer Sertoli cells than controls at all ages studied, while the numbers of Sertoli cells in rats irradiated prenatally were higher than those in controls on day 21. The number of spermatogenic cells was usually decreased in rats irradiated postnatally. In the rats irradiated prenatally normal numbers of spermatogenic cells were found at day 51. Numbers of spermatogenic cells were significantly correlated with the number of Sertoli cells at the ages of 51 and 81 days. The concentration of FSH in the plasma usually increased in the postnatally irradiated animals on days 21 and 51, but not on day 81; prenatal irradiation did not result in altered levels of FSH at any age. Peripheral levels of LH and testosterone were not affected by irradiation. The concentration of FSH in the plasma was negatively correlated with the number of Sertoli cells in all age groups, whereas significant correlations between the level of FSH and the number of spermatogenic cells were only found at days 51 and 81. It is concluded from these data that the Sertoli cell is the most likely source of inhibin.

scholarly journals Pachytene spermatocyte and round spermatid binding to Sertoli cells in vitro

1988 ◽  
Vol 90 (1) ◽  
pp. 105-114
Author(s):  
G.C. Enders ◽  
C.F. Millette
Keyword(s):  
Sertoli Cell ◽  
Sertoli Cells ◽  
Round Spermatid ◽  
Pachytene Spermatocyte ◽  
Assay System ◽  
Spermatogenic Cells ◽  
Vitro Assay ◽  
Cell Monolayers ◽  
Pachytene Spermatocytes

Spermatogenic cells differentiate in vivo while in continuous contact with the Sertoli cell. During differentiation, the spermatogenic cells and Sertoli cells form a number of morphologically distinct stage-specific adhesions. We describe an in vitro assay system for studying the adhesion of spermatogenic cells to Sertoli cell monolayers. Mixed populations of spermatogenic cells or enriched fractions of pachytene spermatocytes and round spermatids were labelled with the vital dye, fluorescein diacetate, prior to their addition to Sertoli cell monolayers so that the adhesion of viable spermatogenic cells could be quantified. Using this assay system, the number of pachytene spermatocyte and round spermatid binding sites on the Sertoli cell monolayer were similar, but the kinetics of binding were different. Pachytene spermatocytes were able to inhibit significantly round spermatid binding, while round spermatids did not significantly inhibit pachytene spermatocyte binding. After coculture for 24–48 h, spermatocytes form junctional structures with Sertoli cells that are similar to desmosome-like junctions. These results suggest that pachytene spermatocytes and round spermatids bind to Sertoli cells by different mechanisms.

scholarly journals Proliferative Phase Sertoli Cells Display a Developmentally Regulated Response to Activin in Vitro

Endocrinology ◽  
2003 ◽  
Vol 144 (2) ◽  
pp. 474-483 ◽  
Author(s):  
Jeremy J. Buzzard ◽  
Paul G. Farnworth ◽  
David M. de Kretser ◽  
Anne E. O’Connor ◽  
Nigel G. Wreford ◽  
...  
Keyword(s):  
Sertoli Cell ◽  
Sertoli Cells ◽  
Activin A ◽  
Paracrine Factor ◽  
Binding Studies ◽  
Developmentally Regulated ◽  
Rat Pups ◽  
Peritubular Cells

We have used cultures of highly purified, proliferating rat Sertoli cells collected from d 3, 6, and 9 rat pups to investigate the role of activin A on Sertoli cell division. These studies demonstrate that activin A acts directly on d 6 and 9, but not d 3, Sertoli cells to induce proliferation, both alone and synergistically with FSH. In addition to stimulating proliferation, activin A induces secretion of inhibins A and B as determined by specific ELISAs. We demonstrate that the synergy between activin A and FSH is not due to local actions of secreted inhibin or follistatin. We have used real-time fluorometric RT-PCR to demonstrate that activin regulates expression of activin receptor and follistatin mRNA by Sertoli cells. Saturation binding studies using 125I-activin A indicate that synergy between activin and FSH may be due to increased numbers of activin receptors on the Sertoli cell. Finally, we show that activin A was secreted at high levels by cultured peritubular cells but was undetectable in high purity proliferating Sertoli cell cultures, suggesting that activin A functions as a paracrine factor during postnatal testis development.

Endocrine control of testicular somatic and premeiotic germ cell development in the immature testis of the primate Macaca mulatta

1995 ◽  
Vol 133 (2) ◽  
pp. 235-247 ◽  
Author(s):  
Stefan Schlatt ◽  
Muhammad Arslan ◽  
Gerhard F Weinbauer ◽  
Hermann M Behre ◽  
Eberhard Nieschlag
Keyword(s):  
Sertoli Cell ◽  
Macaca Mulatta ◽  
Sertoli Cells ◽  
Leydig Cells ◽  
Combined Treatment ◽  
Morphological Differentiation ◽  
Hormone Treatment ◽  
Endocrine Control ◽  
Peritubular Cells ◽  
Spermatogonial Proliferation

Schlatt S, Arslan M, Weinbauer GF, Behre HM, Nieschlag E. Endocrine control of testicular somatic and premeiotic germ cell development in the immature testis of the primate Macaca mulatta. Eur J Endocrinol 1995:133:235–47. ISSN 0804–4643 Four groups (N = 3 per group) of juvenile rhesus monkeys (Macaca mulatta, 14–20 months old) received either vehicle or highly purified human follicle-stimulating hormone (FSH; 10 IU kg−1 day−1), human chorionic gonadotropin (hCG; 250 IU every alternate day) or both hormones for a period of 4 weeks. Testicular volume and weight increased more than twofold after single and more than sixfold after combined hormone treatment. Serum and intratesticular testosterone were at supraphysiological levels in hCG-treated animals and rose even more after combined treatment; a minor elevation of intratesticular testosterone was also observed after FSH treatment. Serum inhibin was elevated after hCG or FSH treatment and increased more than twofold during the first 3 weeks of combined treatment. Semiquantitative analysis of cell numbers showed a statistically non-significant increase in Sertoli cells and Ad- and Ap-spermatogonia after single hormone treatment. Combined treatment induced a further increase in the number of spermatogonia. Leydig cells were only encountered after hCG treatment; their number was more than threefold higher after combined treatment compared with hCG alone. Follicle-stimulating hormone stimulated Sertoli cell and Ap spermatogonia proliferation but did not induce morphological differentiation of Sertoli cells, peritubular cells or Leydig cells. Human CG treatment, however, induced Sertoli cell proliferation and morphological differentiation. It had effects on spermatogonial proliferation but induced differentiation of peritubular cells. Combined treatment initiated the greatest morphological and functional differentiation of Sertoli cells, peritubular cells, Leydig cells and spermatogonia. Flow cytometric analysis confirms an increase of mitotically active cells. The observations show that FSH and testosterone can induce Sertoli cell proliferation. Morphological differentiation of Sertoli cells may be mediated indirectly by environmental and paracrine stimuli released from peritubular cells, whose differentiation is androgen dependent. Leydig cells are stimulated mainly by hCG. Our present and previous data lead us to propose that FSH contributes to the final number and activity of Leydig cells, which secrete immunoreactive inhibin in response to hCG. Spermatogonial proliferation is under the control of FSH, whereas the survival of germ cells is dependent on Sertoli cell function. The observed rise in the number of mitotically inactive Ad-spermatogonia can be explained by a transformation of Apspermatogonia into resting Ad-spermatogonia. E Nieschlag. Institute of Reproductive Medicine of the University, Steinfurter Str. 107, D-48149 Münster, Germany

scholarly journals Electron Microscopic Observations on Mitochondria of Rat Testes Fixed in Potassium Permanganate

1960 ◽  
Vol 7 (2) ◽  
pp. 311-314 ◽  
Author(s):  
William Zebrun ◽  
Hilton H. Mollenhauer
Keyword(s):  
Epithelial Cells ◽  
Potassium Permanganate ◽  
Apparent Density ◽  
Sertoli Cells ◽  
Structural Difference ◽  
Cell Types ◽  
Morphological Investigation ◽  
Spermatogenic Cells ◽  
Mitochondrial Membranes ◽  
Electron Microscopic

A morphological investigation of mitochondria within the seminal epithelial cells of rat testes fixed in potassium permanganate reveals differences in electron opacity between the internal mitochondrial membranes of spermatogenic cells and those of Sertoli cells. Some interpretations of the apparent density differences are briefly discussed. It is concluded that the different effects of permanganate fixation upon the mitochondria of these cell types may reflect a significant structural difference between them.

Sign in / Sign up

Export Citation Format

Share Document