CONVERSION OF PROGESTERONE TO ANDROGENS BY NON-FLAGELLATE GERMINAL CELLS ISOLATED FROM SEMINIFEROUS TUBULES OF RAT TESTIS

1974 ◽  
Vol 60 (2) ◽  
pp. 269-NP ◽  
Author(s):  
H. J. GALENA ◽  
C. TERNER
Keyword(s):  
Significant Contribution ◽  
Isotonic Saline ◽  
Interstitial Cells ◽  
Seminiferous Tubules ◽  
Distilled Water ◽  
Rat Testis ◽  
Germinal Cells

SUMMARY A method is described for the isolation of non-flagellate germinal cells of the testis. The interstitial cells were removed by submersion of teased seminiferous tubules in distilled water. The interstitial cells exposed to water burst while the germinal cells inside the tubules remained intact. The tubules were then homogenized in isotonic saline and the non-flagellate germinal cells (spermatocytes and young spermatids) were isolated by centrifugation and filtration through a layer of Sephadex G-25 gel. On incubation with progesterone these cells produced 17α-hydroxyprogesterone, androstenedione, and testosterone. The rate of conversion of progesterone to testosterone in vitro was 0·20 μg/h/109 germinal cells. These results suggest that the non-flagellate germinal cells of the testis may make a significant contribution to the production of androgens.

scholarly journals The Effect of Cryptorchidism on the Quantitative Histology, Histochemistry and Hydrolytic Enzyme Activity of the Rat Testis

1978 ◽  
Vol 31 (1) ◽  
pp. 53 ◽  
Author(s):  
AW Blackshaw ◽  
PF Massey
Keyword(s):  
Enzyme Activity ◽  
Meiotic Prophase ◽  
Hydrolytic Enzyme ◽  
Seminiferous Tubules ◽  
Quantitative Histology ◽  
Rat Testis ◽  
Germinal Cells ◽  
Enzyme Patterns

Cryptorchidism of the mature rat testis led to degeneration of the seminiferous tubules and changes in enzyme patterns and activities. Spermatogenic stages 1--4, containing pachytene primary spermatocytes in late meiotic prophase, and stage 5, containing recently formed round spermatids, were damaged by 48 h. Within 96 h stages showed a loss of germinal cells into the lumen and this was almost complete by 192 h.

scholarly journals Molecular Cloning and Expression of a Functionally Different Alternative Splice Variant of Prointerleukin-1α from the Rat Testis*

Endocrinology ◽  
2000 ◽  
Vol 141 (12) ◽  
pp. 4413-4418 ◽  
Author(s):  
Taranum Sultana ◽  
Konstantin Svechnikov ◽  
Günther Weber ◽  
Olle Söder
Keyword(s):  
Alternative Splice ◽  
Splice Variant ◽  
Messenger Rna ◽  
Constitutive Expression ◽  
Cloning And Expression ◽  
Seminiferous Tubules ◽  
Alternative Splice Variant ◽  
Rat Testis ◽  
Dose Dependent

Abstract We report here the characterization of an alternative splice variant of prointerleukin-1α (proIL-1α), constitutively expressed by the normal adult rat testis. In addition to the classical 32K proIL-1α (32proIL-1α) messenger RNA, the testis produced a shorter variant encoding a putative protein of 24K (24proIL-1α). In situ hybridization demonstrated constitutive expression of the splice transcript in the seminiferous tubules. This alternative complementary DNA lacked the fifth exon, harboring the calpain cleavage site essential for generation of mature 17K IL-1α. This was verified by calpain treatment, producing the expected cleavage products of recombinant 32proIL-1α, but not of 24proIL-1α. Similarly, expression in COS-7 cells demonstrated processing of 32proIL-1α to the mature 17K form and secretion, whereas 24proIL-1α remained unprocessed. Both 32proIL-1α and 24proIL-1α showed a dose-dependent stimulatory effect in a thymocyte proliferation assay, although at lower potency than mature 17K IL-1α. In contrast, when tested on hCG-stimulated Leydig cells in vitro, a dose-dependent inhibition of testosterone production was obtained with mature 17K IL-1α and at a lower potency with 32proIL-1α, whereas 24proIL-1α was inactive. In conclusion, the three IL-1 bioactive proteins described here contribute to IL-1 protein heterogeneity and may serve as constitutive paracrine mediators in the testis.

Ethan-1,2-dimethanesulphonate reduces testicular oxytocin content and seminiferous tubule movements in the rat

1987 ◽  
Vol 112 (2) ◽  
pp. 311-NP ◽  
Author(s):  
H. D. Nicholson ◽  
R. T. S. Worley ◽  
S. E. F. Guldenaar ◽  
B. T. Pickering
Keyword(s):  
Leydig Cell ◽  
Seminiferous Tubule ◽  
Leydig Cells ◽  
Contractile Activity ◽  
Histological Study ◽  
Interstitial Cells ◽  
Seminiferous Tubules ◽  
Adult Rats ◽  
Adult Rat

ABSTRACT An oxytocin-like peptide is present in the interstitial cells of the testis, and testicular concentrations of oxytocin have been shown to increase seminiferous tubule movements in vitro. We have used the drug ethan-1,2-dimethanesulphonate (EDS), which depletes the Leydig cell population of the adult rat testis, to examine further the relationships between the Leydig cell, testicular oxytocin and tubular movements. Adult rats were injected i.p. with a single dose of EDS (75 mg/kg) or of vehicle (25% dimethyl sulphoxide). Histological study 3 and 10 days after treatment with EDS showed a reduction in the number of interstitial cells, and levels of oxytocin immunoreactivity were undetectable by radioimmunoassay. Immunostaining revealed very few oxytocin-reactive cells. Spontaneous contractile activity of the seminiferous tubules in vitro was also dramatically reduced, but could be restored by the addition of oxytocin to the medium. Four weeks after EDS treatment, the interstitial cells were similar to those in the control animals both in number and in immunostaining; immunoassayable oxytocin was present and tubular movements were normal. The EDS effect, seen at 3 and 10 days, was not altered by daily treatment with testosterone. However, repopulation of the testes with oxytocin-immunoreactive cells was not seen until 6 weeks in the testosterone-treated animals. We suggest that the Leydig cells are the main source of oxytocin immunoreactivity in the testis and that this oxytocin is involved in modulating seminiferous tubule movements and the resultant sperm transport. The results also imply that testosterone does not play a major role in controlling tubular activity in the mature rat. J. Endocr. (1987) 112, 311–316

Growth and Observations of Chinese Hamster Seminiferous Epithelium in Vitro

1970 ◽  
Vol 6 (1) ◽  
pp. 195-205
Author(s):  
D. J. ELLINGSON ◽  
K. T. S. YAO
Keyword(s):  
Cell Formation ◽  
Chinese Hamster ◽  
Time Lapse ◽  
Seminiferous Tubules ◽  
Chinese Hamsters ◽  
Cell Association ◽  
Nuclear Rotation ◽  
Dialysis Membranes ◽  
Germinal Cells

Seminiferous tubules from 1- to 3.5-month-old Chinese hamsters were cultivated under dialysis membranes in Rose chambers. The growth and development of the germinal cells was followed daily with phase-contrast microscopy and time-lapse cinemicrography. Spermatogonia lived for 2 or 3 weeks and underwent frequent mitoses. Spermatocytes in metaphase at culture initiation completed their meiotic division. These cells remained healthy for 3-4 days. Such phenomena as germinal cell/Sertoli cell association, nuclear rotation, multinucleated cell formation and spermatid formation were studied and photographed.

scholarly journals The expression and regulation of Bcl-2-related ovarian killer (Bok) mRNA in the developing and adult rat testis

2001 ◽  
pp. 771-778 ◽  
Author(s):  
JS Suominen ◽  
W Yan ◽  
J Toppari ◽  
A Kaipia
Keyword(s):  
Mrna Expression ◽  
Northern Hybridization ◽  
Seminiferous Tubules ◽  
Testicular Development ◽  
Rat Testis ◽  
Adult Rat ◽  
Before And After ◽  
Hybridization Intensity ◽  
Pachytene Spermatocytes

OBJECTIVE: To study the role of Bcl-2-related ovarian killer (Bok) in the regulation of apoptosis in the testis of developing and adult rat. METHODS: Bok mRNA expression was analyzed by Northern hybridization before and after culturing rat seminiferous tubules in vitro. Seminiferous tubules were cultured with different hormones and growth factors. Changes in the expression level of Bok mRNA during testicular development was analyzed by Northern hybridization. Localization of Bok mRNA was verified by in situ hybridization. RESULTS: Bok mRNA was highly expressed in the rat testis, varying during development. Highest expression levels were found in immature rats. Highest hybridization intensity appeared to be in spermatogonia, pachytene spermatocytes and Sertoli cells. Treatment with FSH was able to inhibit spontaneous increase of Bok mRNA expression that occurred in the defined stages of the rat seminiferous epithelium. CONCLUSIONS: FSH protects germ cells from apoptosis and this protective effect may at least partly be due to the inhibition of Bok gene expression. The amount of apoptosis varies during testicular development and highest expression of Bok mRNA occurs at the time of apoptosis, suggesting a possible role for Bok in its regulation.

Coordinated expression of UT-A and UT-B urea transporters in rat testis

2002 ◽  
Vol 282 (6) ◽  
pp. C1492-C1501 ◽  
Author(s):  
R. A. Fenton ◽  
G. J. Cooper ◽  
I. D. Morris ◽  
C. P. Smith
Keyword(s):  
Sertoli Cell ◽  
Seminiferous Tubule ◽  
Northern Analysis ◽  
Seminiferous Tubules ◽  
Cell Nuclei ◽  
Rat Testis ◽  
Selective Permeability ◽  
Coordinated Expression ◽  
Stage Viii

The blood-seminiferous tubule barrier is responsible for maintaining the unique microenvironment conducive to spermatogenesis. A key feature of the blood-testis barrier is selective permeability to solutes and water transport, conferred by the Sertoli cells of the seminiferous tubules (SMTs). Movement of fluid into the lumen of the seminiferous tubule is crucial to spermatogenesis. By Northern analysis, we have shown that 4.0-, 3.3-, 2.8-, and ∼1.7-kb UT-A mRNA transcripts and a 3.8-kb UT-B mRNA transcript are detected within rat testis. Western analysis revealed the expression of both characterized and novel UT-A and UT-B proteins within the testis. Immunolocalization studies determined that UT-A and UT-B protein expression are coordinated with the developmental stage of the SMT. UT-A proteins were detected in Sertoli cell nuclei at all stages of tubule development and in residual bodies of stage VIII tubules. UT-B protein was expressed on Sertoli cell membranes of stage II–III tubules. Using in vitro perfusion, we determined that a phloretin-inhibitable urea pathway exists across the SMTs of rat testis and conclude that UT-B is likely to participate in this pathway.

scholarly journals Induction of tolerance in vitro by autologous murine testicular cells.

1980 ◽  
Vol 151 (4) ◽  
pp. 827-838 ◽  
Author(s):  
U Hurtenbach ◽  
F Morgenstern ◽  
D Bennett
Keyword(s):  
T Cells ◽  
Germ Cells ◽  
Lymphocyte Proliferation ◽  
Interstitial Cells ◽  
Seminiferous Tubules ◽  
Testicular Cells ◽  
Self Tolerance ◽  
Lymphoid Cell Population ◽  
Reactive Lymphocytes

We have investigated the regulation of self tolerance in mice by examining lymphocyte reactivity in vitro against two subpopulations of autologous testicular cells: germ cells that were derived from the seminiferous tubules, and interstitial somatic cells. In the presence of germ cells, lymphocyte proliferation was strongly reduced. In contrast, somatic interstitial cells stimulated lymphocyte proliferation. In both cases, reactive lymphocytes were mostly T cells. Suppressor T cells activated by autologous germ cells were nonspecific and capable of inhibiting lymphocyte proliferation against autologous and allogeneic somatic testicular cells as well as against allogeneic spleen cells. Suppression was abrogated after treatment of the responder lymphocytes with anti-Ly-2.2 serum plus complement. Lymphocyte proliferation by autologous interstitial cells was considerably reduced, but not completely abolished, by complement-dependent lysis with anti-Thy-1.2 serum. This may indicate the participation in proliferation of a lymphoid cell population other than T cells.

Mechanism of action of the factor(s) secreted by rat seminiferous tubules and inhibiting interstitial cell testosterone production in vitro

1988 ◽  
Vol 119 (3) ◽  
pp. 427-434 ◽  
Author(s):  
V. Syed ◽  
S. A. Khan ◽  
M. Lindh ◽  
E. M. Ritzén
Keyword(s):  
Molecular Weight ◽  
Adenylate Cyclase ◽  
Regulatory Protein ◽  
Interstitial Cells ◽  
Seminiferous Tubules ◽  
Specific Factor ◽  
Gel Chromatography ◽  
Rat Tissues ◽  
Testosterone Production

Abstract. Rat seminiferous tubules secrete a factor which inhibits LH-dependent steroidogenesis by interstitial cells. The inhibitory activity was found to be specific for the testes, as cytosols from other rat tissues such as the kidney, heart, spleen, liver and epididymis had no significant effect on testosterone production by interstitial cells. Preliminary characterization by Ultrogel AcA 44 gel chromatography demonstrated that the active substance in SMST has a molecular weight between 40–50 kD. Spent medium from incubation of seminiferous tubules (SMST) caused a dose-dependent inhibition of LH-or cholera toxin-stimulated in vitro testosterone production by rat interstitial cells. However, SMST failed to inhibit forskolin-stimulated steroidogenesis. The effect of SMST was not altered by pre-incubating the cells with the sulfhydryl reagent, N-ethylmaleimide. Considering the proposed mode of action of these modulators of adenylate cyclase activity, the present studies suggest that a high molecular weight testis specific factor acts through the guanine nucleotide-binding stimulatory regulatory protein of the adenylate cyclase complex to inhibit LH-dependent testosterone production by Leydig cells.

In vitro development of seminiferous tubules from immature rat testis: ultrastructural and morphometric studies

1984 ◽  
Vol 50 (2) ◽  
pp. 191-194
Author(s):  
J. L. Gelly ◽  
J. L. Delongeas ◽  
E. Barrat ◽  
R. Hatier ◽  
G. Grignon
Keyword(s):  
Seminiferous Tubules ◽  
Immature Rat ◽  
In Vitro Development ◽  
Rat Testis ◽  
Vitro Development
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