Effects of cyclic AMP and phorbol ester on transepithelial electrical resistance of Sertoli cell monolayers in two-compartment culture

Janecki, Andrzej; Jakubowiak, Andrzej; Steinberger, Anna

doi:10.1016/0303-7207(91)90009-h

Cited by 48 publications

(39 citation statements)

References 22 publications

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“…In the current study, we have used a well-established in vitro model of Sertoli cells in bicameral culture (Hadley et al 1987, Handelsman et al 1989, Onoda et al 1990, Janecki et al 1991a, Djakiew & Onoda 1993 to study the effects of androgens and FSH on TJs. TJ function in this culture system is readily monitored by the determination of transepithelial electrical resistance (TER; Janecki et al 1991a, 1991b, Chung et al 1999, Fanning et al 1999, Li et al 2001, Lui et al 2003a, Siu et al 2003. Testosterone and FSH, either alone or in combination, are known to increase rat Sertoli cell TER two to threefold in this model (Janecki et al 1991a, 1991b, Steinberger & Klinefelter 1993 indicating that Sertoli cell TJ function can be regulated by gonadotrophins.…”

Section: Introductionmentioning

confidence: 99%

“…TJ function in this culture system is readily monitored by the determination of transepithelial electrical resistance (TER; Janecki et al 1991a, 1991b, Chung et al 1999, Fanning et al 1999, Li et al 2001, Lui et al 2003a, Siu et al 2003. Testosterone and FSH, either alone or in combination, are known to increase rat Sertoli cell TER two to threefold in this model (Janecki et al 1991a, 1991b, Steinberger & Klinefelter 1993 indicating that Sertoli cell TJ function can be regulated by gonadotrophins. Hence, the aim of this study was to further elucidate the hormonal regulation of key TJ components (claudin-11, claudin-3, occludin) by correlating the effects of steroid (T, DHT and estradiol (E2)) treatment and FSH with both the functional capacity of TJs (by monitoring TER), and with TJ protein mRNA expression and immunolocalisation.…”

Section: Introductionmentioning

confidence: 99%

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Claudin-11 expression and localisation is regulated by androgens in rat Sertoli cells in vitro

Kaitu’u-Lino¹,

Sluka²,

Foo³

et al. 2007

Reproduction

125

111

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Claudin-11 and occludin are protein components in tight junctions (TJs) between Sertoli cells which are important for the maintenance of the blood-testis barrier. Barrier formation occurs during puberty, with evidence suggesting hormonal regulation of both claudin-11 and occludin. This study aimed to investigate the regulation of claudin-11 and occludin mRNA expression by testosterone (T) and FSH and their immunolocalisation at rat Sertoli cell TJs in vitro, and to correlate any steroid regulation with the functional capacity of TJs. Sertoli cells formed functional TJs within 3 days as assessed by transepithelial electrical resistance (TER). Both T and dihydrotestosterone significantly (P!0.01) increased TER twofold and claudin-11 mRNA two-to threefold within 3 days. FSH partially stimulated TER and claudin-11 mRNA, but estradiol had no effect. T also promoted claudin-11 localisation into extensive inter-cellular contacts. In contrast to claudin-11, T and FSH did not change occludin mRNA expression, however, T promoted localisation of occludin at cell contacts in a similar manner to claudin-11. Addition of flutamide to T-stimulated cells caused a twofold decrease in both TER and claudin-11 mRNA expression, and resulted in the loss of both proteins from cell contacts. This effect was reversible following flutamide removal. It is concluded that androgens i) co-regulate claudin-11 mRNA expression and TER, implicating claudin-11 in TJ formation and ii) promote the localisation of claudin-11 and occludin at Sertoli cell contacts. Hence, the ability of androgens to maintain spermatogenesis in vivo is partly via their effects on TJ proteins and regulation of the blood-testis barrier.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Claudin-11 expression and localisation is regulated by androgens in rat Sertoli cells in vitro

Kaitu’u-Lino¹,

Sluka²,

Foo³

et al. 2007

Reproduction

125

111

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“…Both follicle-stimulating hormone (FSH) and testosterone increase the function of rat Sertoli cell TJs in vitro (Janecki et al 1991, Kaitu'u-Lino et al 2007 in part by up-regulating claudin-11 mRNA and protein (Kaitu'u-Lino et al 2007), although FSH has also been found to down-regulate claudin-11 mRNA expression in cultured mouse Sertoli cells (Hellani et al 2000). In vivo, selective knockout of the Sertoli cell androgen receptor in mice results in a limited reduction in claudin-11 protein (Tan et al 2005(Tan et al 2005, but a substantial tenfold decrease in claudin-3 gene transcription (Meng et al 2005).…”

Section: Introductionmentioning

confidence: 99%

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

Tarulli¹,

Meachem²,

Schlatt³

et al. 2008

Reproduction

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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.

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“…A widely used in vitro system was used in this study for isolation and culturing of Sertoli cells as described (Byers et al 1986, Janecki et al 1991a,b, 1992, Okanlawon & Dym 1996, Grima et al 1998, Lui et al 2001. Sertoli cells were isolated from 20-day-old male Sprague Dawley rats (CMU) and cultured in serumfree DMEM/F12 (Gibco) at 35 8C in a humidified atmosphere of 5% CO 2 with epidermal growth factor, insulin, transferrin, and bacitracin as described .…”

Section: Primary Sertoli Cell Culturesmentioning

confidence: 99%

RAB13 regulates Sertoli cell permeability barrier dynamics through protein kinase A

Su¹,

Liu

2013

Journal of Molecular Endocrinology

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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 cells in vitro with an established tight junction permeability barrier that mimics the BTB in 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 of Rab13 was 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 and Rab13 knockdown 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.

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Effects of cyclic AMP and phorbol ester on transepithelial electrical resistance of Sertoli cell monolayers in two-compartment culture

Cited by 48 publications

References 22 publications

Claudin-11 expression and localisation is regulated by androgens in rat Sertoli cells in vitro

Claudin-11 expression and localisation is regulated by androgens in rat Sertoli cells in vitro

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

RAB13 regulates Sertoli cell permeability barrier dynamics through protein kinase A

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