231. Sertoli cells de-differentiate in men after chronic gonadotrophin suppression

2008 ◽  
Vol 20 (9) ◽  
pp. 31
Author(s):  
G. A. Tarulli ◽  
P. G. Stanton ◽  
K. Loveland ◽  
E. Rajpert De Meyts ◽  
R. I. McLachlan ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Tight Junction ◽  
Testicular Cancer ◽  
Sertoli Cells ◽  
Junction Protein ◽  
Associated Proteins ◽  
Protein Localisation ◽  
Differentiation Status ◽  
Normal Men ◽  
State 1

Recent studies suggest that adult Sertoli cells are not part of a homogenous and terminally differentiated population in phenotypes infertility1–4. The aims of this study were to compare the differentiation status of Sertoli cells from normal men undergoing gonadotrophin suppression by hormonal-based contraception (androgen plus progestin), as well as in pre-malignant and malignant testicular cancer. Confocal microscopy was performed to assess the expression of markers of cell proliferation (PCNA) and differentiation (androgen receptor) in Sertoli cells in all tissues. As additional markers of differentiation, Sertoli cell tight junction (claudin-11, JAM-A) and associated proteins (ZO-1) were assessed in men with testicular cancer. Samples from five different men were assessed in each group. In normal men, Sertoli cells exhibited a differentiated phenotype (i.e. PCNA negative, androgen receptor positive). However, after gonadotrophin suppression, 1.7 ± 0.6% of Sertoli cells exhibited intense PCNA reactivity and a reduction in androgen receptor immunoreactivity, demonstrating an undifferentiated phenotype. PCNA-positive Sertoli cells were never observed in pre-malignancy, and were only rarely observed in malignant testicular cancer, indicating a potential change in differentiation. Tight junction protein localisation was disrupted in pre-malignant cancer, with a reduction in JAM-A reactivity in Sertoli cells from pre-malignancy and strong JAM-A reactivity in malignant cancer; suggesting a potential role for JAM-A expression in the progression of testicular cancer. We conclude that Sertoli cells are not a homogenous and terminally differentiated population in men and their differentiation is modifiable by hormones and in the disease state. (1) Meachem et al. 2005, Biol Reprod. 72:1187–93 (2) Tarulli et al. 2006, Biol Reprod. 74:798–806 (3) Donner et al. 2004, APMIS. 112:79–88 (4) Brehm et al. 2006, Anat Embryol (Berl). 211(3):223–36

scholarly journals Tetrapeptides Modelled to the Androgen Binding Site of ZIP9 Stimulate Expression of Tight Junction Proteins and Tight Junction Formation in Sertoli Cells

Biology ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 55
Author(s):  
Marie-Louise Möller ◽  
Ahmed Bulldan ◽  
Georgios Scheiner-Bobis
Keyword(s):  
Androgen Receptor ◽  
Tight Junction ◽  
Binding Site ◽  
Sertoli Cells ◽  
Alternative Pathway ◽  
Tight Junction Formation ◽  
Associated Proteins ◽  
A Cell ◽  
Androgen Binding

Androgens stimulate the expression of tight junction (TJ) proteins and the formation of the blood–testis barrier (BTB). Interactions of testosterone with the zinc transporter ZIP9 stimulate the expression of TJ-forming proteins and promote TJ formation in Sertoli cells. In order to investigate androgenic effects mediated by ZIP9 but not by the nuclear androgen receptor (AR), the effects of three tetrapeptides fitting the androgen binding site of ZIP9 were compared with those induced by testosterone in a Sertoli cell line expressing ZIP9 but not the AR. Three tetrapeptides and testosterone displaced testosterone-BSA-FITC from the surface of 93RS2 cells and stimulated the non-classical testosterone signaling pathway that includes the activation of Erk1/2 kinases and transcription factors CREB and ATF-1. The expression of the TJ-associated proteins ZO-1 and claudin-5 was triggered as was the re-distribution of claudin-1 from the cytosol to the membrane and nucleus. Furthermore, TJ formation was stimulated, indicated by increased transepithelial electrical resistance. Silencing ZIP9 expression by siRNA prevented all of these responses. These results are consistent with an alternative pathway for testosterone action at the BTB that does not involve the nuclear AR and highlight the significant role of ZIP9 as a cell-surface androgen receptor that stimulates TJ formation.

scholarly journals Regulation of testicular tight junctions by gonadotrophins in the adult Djungarian hamster in vivo

Reproduction ◽  
2008 ◽  
Vol 135 (6) ◽  
pp. 867-877 ◽  
Author(s):  
Gerard A Tarulli ◽  
Sarah J Meachem ◽  
Stefan Schlatt ◽  
Peter G Stanton
Keyword(s):  
Tight Junction ◽  
Adaptor Protein ◽  
Tight Junction Protein ◽  
Mrna Levels ◽  
Tight Junction Proteins ◽  
Djungarian Hamster ◽  
Junction Protein ◽  
Protein Localisation ◽  
Junction Proteins

This study aimed to assess the effect of gonadotrophin suppression and FSH replacement on testicular tight junction dynamics and blood–testis barrier (BTB) organisation in vivo, utilising the seasonal breeding Djungarian hamster. Confocal immunohistology was used to assess the cellular organisation of tight junction proteins and real-time PCR to quantify tight junction mRNA. The effect of tight junction protein organisation on the BTB permeability was also investigated using a biotin-linked tracer. Tight junction protein (claudin-3, junctional adhesion molecule (JAM)-A and occludin) localisation was present but disorganised after gonadotrophin suppression, while mRNA levels (claudin-11, claudin-3 and occludin) were significantly (two- to threefold) increased. By contrast, both protein localisation and mRNA levels for the adaptor protein zona occludens-1 decreased after gonadotrophin suppression. FSH replacement induced a rapid reorganisation of tight junction protein localisation. The functionality of the BTB (as inferred by biotin tracer permeation) was found to be strongly associated with the organisation and localisation of claudin-11. Surprisingly, JAM-A was also recognised on spermatogonia, suggesting an additional novel role for this protein in trans-epithelial migration of germ cells across the BTB. It is concluded that gonadotrophin regulation of tight junction proteins forming the BTB occurs primarily at the level of protein organisation and not gene transcription in this species, and that immunolocalisation of the organised tight junction protein claudin-11 correlates with BTB functionality.

scholarly journals Tight and Adherens Junctions in the Ovine Uterus: Differential Regulation by Pregnancy and Progesterone

Endocrinology ◽  
2007 ◽  
Vol 148 (8) ◽  
pp. 3922-3931 ◽  
Author(s):  
M. Carey Satterfield ◽  
Kathrin A. Dunlap ◽  
Kanako Hayashi ◽  
Robert C. Burghardt ◽  
Thomas E. Spencer ◽  
...  
Keyword(s):  
Tight Junction ◽  
Adherens Junctions ◽  
Luminal Epithelium ◽  
Tissue Fluid ◽  
Tight Junction Formation ◽  
Junction Protein ◽  
Conceptus Development ◽  
Associated Proteins ◽  
Continuous Presence ◽  
D 12

In species with noninvasive implantation by conceptus trophectoderm, fetal/maternal communications occur across the endometrial epithelia. The present studies identified changes in junctional complexes in the ovine endometrium that regulate paracellular trafficking of water, ions, and other molecules, and the secretory capacity of the uterine epithelia. Distinct temporal and spatial alterations in occludin, tight junction protein 2, and claudin 1–4 proteins were observed in the endometrium of cyclic and early pregnant ewes. Dynamic changes in tight junction formation were characterized by an abundance of tight junction proteins on d 10 of the estrous cycle and pregnancy that substantially decreased by d 12. Early progesterone administration advanced conceptus development on d 9 and 12 that was associated with loss of tight-junction-associated proteins. Pregnancy increased tight-junction-associated proteins between d 14–16. Cadherin 1 and β-catenin, which form adherens junctions, were abundant in the endometrial glands, but decreased after d 10 of pregnancy in the luminal epithelium and then increased by d 16 with the onset of implantation. Results support the ideas that progesterone elicits transient decreases in tight and adherens junctions in the endometrial luminal epithelium between d 10–12 that increases selective serum and tissue fluid transudation to enhance blastocyst elongation, which is subsequently followed by an increase in tight and adherens junctions between d 14–16 that may be required for attachment and adherence of the trophectoderm for implantation. The continuous presence of tight and adherens junctions in the uterine glands would allow for vectorial secretion of trophic substances required for conceptus elongation and survival.

scholarly journals The tight junction protein complex undergoes rapid and continuous molecular remodeling at steady state

2008 ◽  
Vol 181 (4) ◽  
pp. 683-695 ◽  
Author(s):  
Le Shen ◽  
Christopher R. Weber ◽  
Jerrold R. Turner
Keyword(s):  
Tight Junction ◽  
Diffusion Constant ◽  
Tight Junction Protein ◽  
Dependent Manner ◽  
Zonula Occludens ◽  
Determine Function ◽  
Junction Protein ◽  
Associated Proteins ◽  
Energy Dependent ◽  
Darby Canine Kidney

The tight junction defines epithelial organization. Structurally, the tight junction is comprised of transmembrane and membrane-associated proteins that are thought to assemble into stable complexes to determine function. In this study, we measure tight junction protein dynamics in live confluent Madin–Darby canine kidney monolayers using fluorescence recovery after photobleaching and related methods. Mathematical modeling shows that the majority of claudin-1 (76 ± 5%) is stably localized at the tight junction. In contrast, the majority of occludin (71 ± 3%) diffuses rapidly within the tight junction with a diffusion constant of 0.011 μm2s−1. Zonula occludens-1 molecules are also highly dynamic in this region, but, rather than diffusing within the plane of the membrane, 69 ± 5% exchange between membrane and intracellular pools in an energy-dependent manner. These data demonstrate that the tight junction undergoes constant remodeling and suggest that this dynamic behavior may contribute to tight junction assembly and regulation.

116. Contribution of claudin-11 to the inter-Sertoli cell tight junction, in vitro

2005 ◽  
Vol 17 (9) ◽  
pp. 72
Author(s):  
M. J. McCabe ◽  
P. G. Stanton
Keyword(s):  
Tight Junction ◽  
Mrna Expression ◽  
Sertoli Cell ◽  
Adhesion Molecule ◽  
Sertoli Cells ◽  
Adherens Junction ◽  
Adherens Junction Protein ◽  
Junction Protein

The inter-Sertoli cell tight junction (TJ) forms the blood testis barrier (BTB) between Sertoli cells and is composed of three major transmembrane proteins: claudin-11, occludin and junctional adhesion molecule. Formation of the BTB occurs during puberty associating with an increase in circulating gonadotrophins. Claudin-11 and occludin are hormonally regulated in vitro although their importance to the function of the TJ is unknown. The aim of this study was to investigate the contribution of claudin-11 to the inter-Sertoli cell TJ in vitro by blocking gene expression using RNA interference. Two claudin-11-specific siRNA fragments were designed for this purpose. Sertoli cells in primary culture formed stable TJs within 5 days as measured by transepithelial electrical resistance (TER). The addition of siRNA for 2 days resulted in a significant (P < 0.01) 55% (mean, SD, n = 4 cultures) decrease in TER along with a major reduction in claudin-11 localisation to the TJ as assessed by immunocytochemistry. The specificity of the siRNA was shown by the presence of extensive immunostaining of occludin and of the adherens junction protein β-catenin in the same treatments. Similarly, claudin-11 mRNA expression significantly (P < 0.01) decreased by 71% (mean, SD, n = 3 cultures) in response to both claudin-11 siRNA fragments. Occludin mRNA expression was not affected. It is concluded that claudin-11 contributes at least 55% to the function of the rat Sertoli cell TJ in vitro. It is hypothesised that the remaining 45% of TJ function can be attributed to other integral proteins, such as occludin and junctional adhesion molecule. It is expected that claudin-11 and other TJ proteins play a pivotal role in the function of the BTB in vivo with potential implications in fertility and contraception.

scholarly journals Androgen-Dependent Sertoli Cell Tight Junction Remodeling Is Mediated by Multiple Tight Junction Components

2014 ◽  
Vol 28 (7) ◽  
pp. 1055-1072 ◽  
Author(s):  
Papia Chakraborty ◽  
F. William Buaas ◽  
Manju Sharma ◽  
Benjamin E. Smith ◽  
Anne R. Greenlee ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Tight Junction ◽  
Tight Junctions ◽  
Sertoli Cell ◽  
Chromatin Immunoprecipitation ◽  
Target Genes ◽  
Altered Expression ◽  
Junction Protein ◽  
Epithelial Compartment ◽  
Androgen Regulation

Sertoli cell tight junctions (SCTJs) of the seminiferous epithelium create a specialized microenvironment in the testis to aid differentiation of spermatocytes and spermatids from spermatogonial stem cells. SCTJs must be chronically broken and rebuilt with high fidelity to allow the transmigration of preleptotene spermatocytes from the basal to adluminal epithelial compartment. Impairment of androgen signaling in Sertoli cells perturbs SCTJ remodeling. Claudin (CLDN) 3, a tight junction component under androgen regulation, localizes to newly forming SCTJs and is absent in Sertoli cell androgen receptor knockout (SCARKO) mice. We show here that Cldn3-null mice do not phenocopy SCARKO mice: Cldn3−/− mice are fertile, show uninterrupted spermatogenesis, and exhibit fully functional SCTJs based on imaging and small molecule tracer analyses, suggesting that other androgen-regulated genes must contribute to the SCARKO phenotype. To further investigate the SCTJ phenotype observed in SCARKO mutants, we generated a new SCARKO model and extensively analyzed the expression of other tight junction components. In addition to Cldn3, we identified altered expression of several other SCTJ molecules, including down-regulation of Cldn13 and a noncanonical tight junction protein 2 isoform (Tjp2iso3). Chromatin immunoprecipitation was used to demonstrate direct androgen receptor binding to regions of these target genes. Furthermore, we demonstrated that CLDN13 is a constituent of SCTJs and that TJP2iso3 colocalizes with tricellulin, a constituent of tricellular junctions, underscoring the importance of androgen signaling in the regulation of both bicellular and tricellular Sertoli cell tight junctions.

scholarly journals Nalp Signalling is Required in Sertoli Cells for Tight-Junction Protein Interaction

2015 ◽  
Vol 42 (1) ◽  
pp. 12-17
Author(s):  
S. Hayrabedyan ◽  
K. Todorova
Keyword(s):  
Innate Immunity ◽  
Tight Junction ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
Cell Biology ◽  
Cell Types ◽  
Protective Role ◽  
Receptor Ligands ◽  
Junction Protein ◽  
Sertoli Cell Line

Summary The present study aims to investigate the NALP3 system and its influence on occludin in Sertoli cells, utilising primary murine cells and adult Sertoli cell line as models. Its main goals are the Sertoli cell biology with possible implications on male reproductive functions. Primary and adult Sertoli cells were transfected with NAPL3 siRNA and treated with NOD1 (ie-DAP) and NOD2 (MDP) receptor ligands. There was positive occludin expression levels on transcript (RT-qPCR) and protein (FCS and Immunofluorescence) levels for both cell types. The innate immunity and tight-junction pathways integration serve a protective role for both testis immune barrier and spermatogenesis compartmentalisation maintained by the very same barrier. This integration also points the way for mechanistic research of the disturbances inflicted during an inflammatory response in testis niche.

scholarly journals RAB13 regulates Sertoli cell permeability barrier dynamics through protein kinase A

2013 ◽  
Vol 50 (3) ◽  
pp. 305-318 ◽  
Author(s):  
Wenhui Su ◽  
Xinchun Liu
Keyword(s):  
Protein Kinase ◽  
Tight Junction ◽  
Protein Kinase A ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
Cell Permeability ◽  
Permeability Barrier ◽  
Rab Gtpases ◽  
Junction Protein ◽  
Kinase A

In mammalian testes, the blood–testis barrier (BTB), created by specialized junctions between Sertoli cells near the basement membrane of the seminiferous epithelium, provides an indispensable immune-privileged microenvironment for spermatid development. However, the BTB must experience restructuring during the epithelial cycle to facilitate the transit of preleptotene spermatocytes upon the testosterone-induced new TJ fibrils forming behind these cells, which is intimately related to the extensive dynamics of junction protein complexes between Sertoli cells. As key regulators of protein traffic, Rab GTPases participate in delivery of proteins between distinct cellular sites and cross talk with proteins that constitute tight junction and adherens junction. Using primarily cultured Sertoli cellsin vitrowith an established tight junction permeability barrier that mimics the BTBin vivo, RAB13 was shown to decrease during the testosterone-induced TJ integrity enhancement, accompanied with an increment in protein kinase A (PKA) activity. Furthermore, knockdown ofRab13was found to resemble the effect of testosterone on Sertoli cell TJ permeability by reinforcing filamentous actin and occludin distribution at the cell–cell interface and promoting the direct interaction between ZO-1 and occludin. Interestingly, the effects of testosterone andRab13knockdown on Sertoli cell epithelium were revealed to be antagonized by PKA activity inhibition. In summary, RAB13 serves as a regulatory component in the assembly and restructuring of the TJ fibrils between adjacent Sertoli cells.

scholarly journals Mechanisms involved in AMPK-mediated deposition of tight junction components to the plasma membrane

2020 ◽  
Vol 318 (3) ◽  
pp. C486-C501
Author(s):  
Jingshing Wu ◽  
Pascal Rowart ◽  
Francois Jouret ◽  
Brandon M. Gassaway ◽  
Vanathy Rajendran ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Tight Junction ◽  
Adherens Junction ◽  
Bound State ◽  
Ampk Activation ◽  
Zonula Occludens ◽  
Adherens Junction Protein ◽  
Junction Protein ◽  
Associated Proteins ◽  
Amp Activated Protein Kinase

AMP-activated protein kinase (AMPK) activation promotes early stages of epithelial junction assembly. AMPK activation in MDCK renal epithelial cells facilitates localization of the junction-associated proteins aPKCζ and Par3 to the plasma membrane and promotes conversion of Cdc42, a key regulator of epithelial polarization and junction assembly, to its active GTP bound state. Furthermore, Par3 is an important regulator of AMPK-mediated aPKCζ localization. Both aPKCζ and Par3 serve as intermediates in AMPK-mediated junction assembly, with inhibition of aPKCζ activity or Par3 knockdown disrupting AMPK’s ability to facilitate zonula occludens (ZO-1) localization. AMPK phosphorylates the adherens junction protein afadin and regulates its interaction with the tight-junction protein zonula occludens-1. Afadin is phosphorylated at two critical sites, S228 (residing within an aPKCζ consensus site) and S1102 (residing within an AMPK consensus site), that are differentially regulated during junction assembly and that exert different effects on the process. Expression of phospho-defective mutants (S228A and S1102A) perturbed ZO-1 localization to the plasma membrane during AMPK-induced junction assembly. Expression of S228A increased the ZO-1/afadin interaction, while S1102A reduced this interaction during extracellular calcium-induced junction assembly. Inhibition of aPKCζ activity also increased the ZO-1/afadin interaction. Taken together, these data suggest that aPKCζ phosphorylation of afadin terminates the ZO-1/afadin interaction and thus permits the later stages of junction assembly.

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