scholarly journals The Non-hormonal Male Contraceptive Adjudin Exerts its Effects via MAPs and Signaling Proteins mTORC1/rpS6 and FAK-Y407

Endocrinology ◽  
2020 ◽  
Vol 162 (1) ◽  
Author(s):  
Lingling Wang ◽  
Ming Yan ◽  
Huitao Li ◽  
Siwen Wu ◽  
Renshan Ge ◽  
...  
Keyword(s):  
Male Infertility ◽  
Germ Cell ◽  
Cell Function ◽  
Adherens Junction ◽  
Protein S ◽  
Signaling Proteins ◽  
Adult Rats ◽  
Male Contraceptive ◽  
Protein Levels ◽  
Ectoplasmic Specialization

Abstract Adjudin, 1-(2,4-dichlorobenzyl)-1H-indazole-3-carbohydrazide (formerly called AF-2364), is a nonhormonal male contraceptive, since it effectively induces reversible male infertility without perturbing the serum concentrations of follicle stimulating hormone (FSH), testosterone, and inhibin B based on studies in rats and rabbits. Adjudin was shown to exert its effects preferentially by perturbing the testis-specific actin-rich adherens junction (AJ) at the Sertoli–spermatid interface known as apical ectoplasmic specialization (apical ES), thereby effectively inducing spermatid exfoliation. Adjudin did not perturb germ cell development nor germ cell function. Also, it had no effects on Sertoli cell–cell AJ called basal ectoplasmic specialization (basal ES), which, together with tight junction constitute the blood-testis barrier (BTB), unless an acute dose of adjudin was used. Adjudin also did not perturb the population of spermatogonial stem cells nor Sertoli cells in the testis. However, the downstream signaling protein(s) utilized by adjudin to induce transient male infertility remains unexplored. Herein, using adult rats treated with adjudin and monitored changes in the phenotypes across the seminiferous epithelium between 6 and 96 h in parallel with the steady-state protein levels of an array of signaling and cytoskeletal regulatory proteins, recently shown to be involved in apical ES, basal ES and BTB function. It was shown that adjudin exerts its contraceptive effects through changes in microtubule associated proteins (MAPs) and signaling proteins mTORC1/rpS6 and p-FAK-Y407. These findings are important to not only study adjudin-mediated male infertility but also the biology of spermatogenesis.

scholarly journals Dynamin II interacts with the cadherin- and occludin-based protein complexes at the blood–testis barrier in adult rat testes

2006 ◽  
Vol 191 (3) ◽  
pp. 571-586 ◽  
Author(s):  
Pearl P Y Lie ◽  
Weiliang Xia ◽  
Claire Q F Wang ◽  
Dolores D Mruk ◽  
Helen H N Yan ◽  
...  
Keyword(s):  
Germ Cell ◽  
Protein Complexes ◽  
Adherens Junction ◽  
Cell Movement ◽  
Cell Loss ◽  
Adult Rat ◽  
Ectoplasmic Specialization ◽  
Stage Viii ◽  
Blood Testis Barrier

In adult rat testes, blood–testis barrier (BTB) restructuring facilitates the migration of preleptotene spermatocytes from the basal to the adluminal compartment that occurs at stage VIII of the epithelial cycle. Structural proteins at the BTB must utilize an efficient mechanism (e.g. endocytosis) to facilitate its transient ‘opening’. Dynamin II, a large GTPase known to be involved in endocytosis, was shown to be a product of Sertoli and germ cells in the testis. It was also localized to the BTB, as well as the apical ectoplasmic specialization (apical ES), during virtually all stages of the epithelial cycle. By co-immunoprecipitation, dynamin II was shown to associate with occludin, N-cadherin, zonula occludens-1 (ZO-1), β-catenin, junctional adhesion molecule-A, and p130Cas, but not nectin-3. An in vivo model in rats previously characterized for studying adherens junction (AJ) dynamics in the testes by adjudin (formerly called AF-2364, 1-(2,4-dichlorobenzyl)-1H-indazole-3-car-hohydrizide) treatment was used in our studies. At the time of germ cell loss from the seminiferous epithelium as a result of adjudin-induced AJ restructuring without disrupting the BTB integrity, a significant decline in the steady-state dynamin II protein level was detected. This change was associated with a concomitant increase in the levels of two protein complexes at the BTB, namely occludin/ZO-1 and N-cadherin/β-catenin. Interestingly, these changes were also accompanied by a significant increase in the structural interaction of dynamin II with β-catenin and ZO-1. β-Catenin and ZO-1 are adaptors that structurally link the cadherin- and occludin-based protein complexes together at the BTB in an ‘engaged’state to reinforce the barrier function in normal testes. However, β-catenin and ZO-1 were ‘disengaged’ from each other but bound to dynamin II during adjudin-induced AJ restructuring in the testis. The data reported herein suggest that dynamin II may assist the ‘disengagement’ of β-catenin from ZO-1 during BTB restructuring. Thus, this may permit the occludin/ZO-1 complexes to maintain the BTB integrity when the cadherin/catenin complexes are dissociated to facilitate germ cell movement.

scholarly journals Semen testis expressed protein 101 and spermatid-specific thioredoxin reductase 3 levels may be biomarkers in infertile male

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Fatma Zehra Erbayram ◽  
Esma Menevse ◽  
Duygu Dursunoglu
Keyword(s):  
Male Infertility ◽  
Biochemical Parameters ◽  
Sperm Concentration ◽  
Thioredoxin Reductase ◽  
Clinical Approach ◽  
Elisa Method ◽  
Infertile Male ◽  
Protein Levels ◽  
Group 2 ◽  
Group 1

Abstract Objectives We aimed to determine the differences between normozoospermic and oligozoospermic individuals according to levels of spermatid-specific thioredoxin reductase 3 (SPTRXR3/STRX3/TXNDC8/TXNRD3) and testis expressed protein 101 (TEX-101), and to evaluate the correlations between spermiogram data and biochemical parameters. Methods The study was carried out at the Andrology Laboratory of Medicine Faculty of Selcuk University. Two groups were designed: Group 1: Normozoospermia (n=40, sperm concentration ≥ 15 million/mL), Group 2: Oligozoospermia; (n=40, sperm concentration < 15 million/mL). Seminal plasma SPTRXR3 and TEX-101 levels were analyzed with ELISA method. Spermiogram analysis was evaluated according to WHO 2010 Kruger criteria. Results TEX-101 protein levels were significantly different in normozoospermia (2.12 ± 0.08 ng/mL) compared to oligozoospermia (1.55 ± 0.04 ng/mL). SPTRXR3 levels (6.98 ± 0.46 ng/mL) were higher in oligozoospermia than normozoospermia (3.07 ± 0.35 ng/mL). Both TEX-101 and SPTRXR3 levels were correlated statistically with most of the spermiogram parameters. Conclusions High SPTRXR3 and low TEX-101 levels may be a biomarker in evaluation of male infertility. The relations between spermiogram parameters indicates that results present a new clinical approach in biology of oligozoospermic male.

Impaired Testicular Function in Patients With Carcinoma-In-Situ of the Testis

1999 ◽  
Vol 17 (1) ◽  
pp. 173-173 ◽  
Author(s):  
Peter Meidahl Petersen ◽  
Aleksander Giwercman ◽  
Steen W. Hansen ◽  
Jørgen G. Berthelsen ◽  
Gedske Daugaard ◽  
...  
Keyword(s):  
Testicular Cancer ◽  
Germ Cell ◽  
Leydig Cell ◽  
Carcinoma In Situ ◽  
Cell Function ◽  
Semen Quality ◽  
Sperm Concentration ◽  
Testicular Dysfunction ◽  
Leydig Cell Function

PURPOSE: To elucidate the biologic association between germ cell neoplasia and testicular dysfunction, through investigation of Leydig cell function and semen quality in men with carcinoma-in-situ (CIS) of the testis. PATIENTS AND METHODS: We examined two groups of men, unilaterally orchidectomized for testicular cancer. Biopsy of the contralateral testis had showed CIS in a group of 24 patients and no evidence of CIS in the other group of 30 patients. Semen quality and serum levels of testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were compared in these two groups of men after orchidectomy but before further treatment for testicular cancer. RESULTS: Significantly higher LH levels (median, 8.1 IU/L v 4.8 IU/L; P < .001) and generally lower testosterone levels (median, 12.5 nmol/L v 15.5 nmol/L; P = .13) were found in the CIS group. The proportion of patients with Leydig cell dysfunction was higher in the group of patients with CIS (11 of 24) than in the group of patients without (two of 30) (P = .01). Sperm concentration and total sperm count were significantly lower (P < .001) in patients with CIS (median, 0.03 × 106/mL and 0.10 × 106, respectively) than in patients without (median, 9.1 × 106/mL and 32 × 106, respectively), whereas the levels of FSH were significantly higher (P < .001) in the former group of men (median, 19.6 IU/L v 9.0 IU/L). CONCLUSION: Not only spermatogenesis but also Leydig cell function is impaired in testes with CIS. This impairment could be due to common factors in the pathogenesis of germ cell neoplasm and testicular dysfunction. Alternatively, CIS cells may have a negative impact on Leydig cell function.

Functional and molecular characterization of NHE3 expression during ontogeny in rat jejunal epithelium

1997 ◽  
Vol 273 (6) ◽  
pp. C1937-C1946 ◽  
Author(s):  
James F. Collins ◽  
Hua Xu ◽  
Pawel R. Kiela ◽  
Jiamin Zeng ◽  
Fayez K. Ghishan
Keyword(s):  
Northern Blot ◽  
Polyclonal Antibodies ◽  
Membrane Vesicle ◽  
Age Groups ◽  
Fold Increase ◽  
Jejunal Mucosa ◽  
Mrna Levels ◽  
Adult Rats ◽  
Intestinal Brush Border Membrane ◽  
Protein Levels

Ontogenic changes occur in intestinal brush-border membrane vesicle (BBMV) Na+/H+exchange activity. The present studies were designed to investigate ontogenic changes in Na+/H+exchanger (NHE) isoform 3 in rat jejunum. pH-dependent Na+ uptake was assayed in four age groups of rats in the presence of 0, 50, or 800 μM HOE-694, a specific NHE inhibitor with differential sensitivities for NHE2 [inhibition constant ( K i) = 5 μM in PS120 fibroblasts] and NHE3 ( K i = 650 μM). Results showed that NHE2 and NHE3 contribute to basal BBMV uptake at all ages. Uptake levels were highest in 6-wk-old rats, lower in adult rats, and lowest in 2-wk-old (suckling) and 3-wk-old (weanling) rats. NHE3 contribution ranged from 92% at 6 wk of age to 59% at 2 and 3 wk of age. NHE3 inhibition by 800 μM HOE-694 was 38–45%. Statistical analysis showed that HOE-694 had a significant effect at both concentrations at all ages and that differences were present between all ages except 2- and 3-wk rats (at all HOE-694 concentrations). Northern blot analyses of jejunal mucosa showed lowest NHE3 mRNA levels in 2-wk animals and higher levels in all other age groups. Polyclonal antibodies were developed against an NHE3 COOH-terminal fusion protein, and antiserum was characterized with NHE3-transfected PS120 cells and by immunohistochemistry. Western blot analyses showed lowest protein levels in 2-wk animals and higher levels in the other ages. Suckling rats were subcutaneously injected with methylprednisone (MP) for 2 days and killed 1 day later. Northern blot analyses showed a twofold increase in NHE3 mRNA expression with MP treatment. Immunoblot analyses showed a 2.5-fold increase in NHE3 immunoreactive protein levels with MP injection. Overall, these data suggest that NHE3 is regulated during ontogeny and that ontogenic changes are most apparent around the time of weaning. Furthermore, the data suggest that NHE3 is regulated at transcriptional and posttranscriptional levels during mammalian intestinal development.

scholarly journals The Hypoxic Testicle: Physiology and Pathophysiology

2012 ◽  
Vol 2012 ◽  
pp. 1-15 ◽  
Author(s):  
Juan G. Reyes ◽  
Jorge G. Farias ◽  
Sebastián Henríquez-Olavarrieta ◽  
Eva Madrid ◽  
Mario Parraga ◽  
...  
Keyword(s):  
Germ Cell ◽  
Testicular Torsion ◽  
Oxygen Supply ◽  
Cell Function ◽  
Cell Damage ◽  
The Body ◽  
Body Core Temperature ◽  
Seminiferous Tubules ◽  
Low Oxygen ◽  
Complex Biological Process

Mammalian spermatogenesis is a complex biological process occurring in the seminiferous tubules in the testis. This process represents a delicate balance between cell proliferation, differentiation, and apoptosis. In most mammals, the testicles are kept in the scrotum 2 to 7°C below body core temperature, and the spermatogenic process proceeds with a blood and oxygen supply that is fairly independent of changes in other vascular beds in the body. Despite this apparently well-controlled local environment, pathologies such as varicocele or testicular torsion and environmental exposure to low oxygen (hypoxia) can result in changes in blood flow, nutrients, and oxygen supply along with an increased local temperature that may induce adverse effects on Leydig cell function and spermatogenesis. These conditions may lead to male subfertility or infertility. Our literature analyses and our own results suggest that conditions such as germ cell apoptosis and DNA damage are common features in hypoxia and varicocele and testicular torsion. Furthermore, oxidative damage seems to be present in these conditions during the initiation stages of germ cell damage and apoptosis. Other mechanisms like membrane-bound metalloproteinases and phospholipase A2 activation could also be part of the pathophysiological consequences of testicular hypoxia.

scholarly journals Phase Transitioned Nuclear Oskar Promotes Cell Division of Drosophila Primordial Germ Cells

2018 ◽  
Author(s):  
Kathryn E. Kistler ◽  
Tatjana Trcek ◽  
Thomas R. Hurd ◽  
Ruoyu Chen ◽  
Feng-Xia Liang ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Fate ◽  
Germ Cells ◽  
Cell Function ◽  
Primordial Germ Cells ◽  
Nuclear Localization Sequence ◽  
Cell Systems ◽  
Germ Granules ◽  
Localization Sequence ◽  
Transcriptional Gene Regulation

ABSTRACTGerm granules are non-membranous ribonucleoprotein granules deemed the hubs for post-transcriptional gene regulation and functionally linked to germ cell fate across species. Little is known about the physical properties of germ granules and how these relate to germ cell function. Here we study two types of germ granules in the Drosophila embryo: cytoplasmic germ granules that instruct primordial germ cells (PGCs) formation and nuclear germ granules within early PGCs with unknown function. We show that cytoplasmic and nuclear germ granules are phase transitioned condensates nucleated by Oskar protein that display liquid as well as hydrogel-like properties. Focusing on nuclear granules, we find that Oskar drives their formation in heterologous cell systems. Multiple, independent Oskar protein domains synergize to promote granule phase separation. Deletion of Oskar’s nuclear localization sequence specifically ablates nuclear granules in cell systems. In the embryo, nuclear germ granules promote germ cell divisions thereby increasing PGC number for the next generation.

Male infertility. More than a statistical risk factor for Germ Cell Tumors?

2000 ◽  
Vol 33 (2) ◽  
pp. 135-139
Author(s):  
H. Büttner ◽  
P. Fornara ◽  
J. Sandmann ◽  
C. Doehn ◽  
D. Jocham
Keyword(s):  
Risk Factor ◽  
Male Infertility ◽  
Germ Cell ◽  
Germ Cell Tumors ◽  
Statistical Risk

The murine seminiferous epithelial cycle is pre-figured in the Sertoli cells of the embryonic testis

Development ◽  
2002 ◽  
Vol 129 (3) ◽  
pp. 635-647 ◽  
Author(s):  
Paula M. Timmons ◽  
Peter W. J. Rigby ◽  
Françoise Poirier
Keyword(s):  
Germ Cell ◽  
Sertoli Cells ◽  
Cell Function ◽  
Seminiferous Epithelium ◽  
Developmental Origins ◽  
Sperm Production ◽  
Functional Heterogeneity ◽  
Adult Testis ◽  
Germ Cell Apoptosis ◽  
Cell Lineages

The seminiferous epithelial cycle and spermatogenic wave are conserved features of vertebrate spermatogenic organisation that reflect the need for the rigorous maintenance of sperm production. Although the cycle and the wave of the adult seminiferous epithelium have been well characterised, particularly in rodent species, their developmental origins are unknown. We show that the Sertoli cells of the pre-pubertal mouse, including those of the germ cell-deficient XXSxra mutant, exhibit coordinated, cyclical patterns of gene expression, presaging the situation in the adult testis, where Sertoli cell function is coupled to the spermatogenic cycle. In the case of the galectin 1 gene (Lgals1), localised differential expression in the Sertoli cells can be traced back to neonatal and embryonic stages, making this the earliest known molecular marker of functional heterogeneity in mammalian testis cords. In addition, the timing of germ cell apoptosis in normal pre-pubertal testes is linked to the temporal cycle of the Sertoli cells. These data show that the cycle and wave of the murine seminiferous epithelium originate at a much earlier stage in development than was previously known, and that their maintenance in the early postnatal cords depends exclusively on the somatic cell lineages.

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