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Article: Transforming growth factor-β3 perturbs the inter-Sertoli tight junction permeability barrier in vitro possibly mediated via its effects on occludin, zonula occludens-1, and claudin-11

TitleTransforming growth factor-β3 perturbs the inter-Sertoli tight junction permeability barrier in vitro possibly mediated via its effects on occludin, zonula occludens-1, and claudin-11
Authors
Issue Date2001
PublisherThe Endocrine Society. The Journal's web site is located at http://endo.endojournals.org
Citation
Endocrinology, 2001, v. 142 n. 5, p. 1865-1877 How to Cite?
AbstractThroughout spermatogenesis, inter-Sertoli tight junctions (TJs) that create the blood-testis barrier in the rat must be disassembled and reassembled to permit the timely passage of preleptotene spermatocytes from the basal to the adluminal compartment of the seminiferous epithelium. However, the mechanism(s) and the participating molecules that regulate this event are largely unknown. Although there is no in vitro model to study the event and regulation of inter-Sertoli TJ disassembly, primary cultures of Sertoli cells in vitro can be used to study junction assembly. In this study, we sought to investigate whether cytokines are involved in the inter-Sertoli TJ assembly in vitro. Sertoli cells isolated from 20-day-old rats were cultured at a density of 0.5-1.2 × 10 6 cells/cm 2 on Matrigel-coated dishes or bicameral units for 8-9 days. The steady-state messenger RNA levels of basic fibroblast growth factor (bFGF), transforming growth factor (TGF)-β2, and TGF-β3 at different time points were assessed by semiquantitative RT-PCR. In selected experiments, the assembly of inter-Sertoli TJs was monitored by transepithelial electrical resistance measurement. It was found that there was no change in the expression of basic fibroblast growth factor throughout the entire culture period. However, there was a 2-fold reduction in the expression of TGF-β2 and TGF-β3 at the time inter-Sertoli TJs were being assembled. On days 5-8, after the inter-Sertoli TJs had been assembled, the Sertoli cell steady-state messenger RNA levels of TGF-β2 and TGF-β3 increased by as much as 3- and 6-fold, respectively, when compared with Sertoli cells on days 1-3 when TJs were being assembled. Also, it was found that recombinant TGF-β3 added to Sertoli cells cultured in vitro at 1.2 × 10 6 cells/cm 2 on Matrigel-coated bicameral units perturbed the inter-Sertoli TJ permeability barrier dose-dependently. Moreover, the presence of TGF-β3 also inhibited the transient and/or basal expression of several TJ-associated proteins, which include occludin, zonula occludens-1, and claudin-11 when inter-Sertoli TJs were being assembled in vitro. These results suggest that TGF-β plays a crucial role in regulating the complicated biochemical events of junction assembly in the testis.
Persistent Identifierhttp://hdl.handle.net/10722/178743
ISSN
2015 Impact Factor: 4.159
2015 SCImago Journal Rankings: 2.363
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLui, WYen_US
dc.contributor.authorLee, WMen_US
dc.contributor.authorCheng, CYen_US
dc.date.accessioned2012-12-19T09:49:26Z-
dc.date.available2012-12-19T09:49:26Z-
dc.date.issued2001en_US
dc.identifier.citationEndocrinology, 2001, v. 142 n. 5, p. 1865-1877en_US
dc.identifier.issn0013-7227en_US
dc.identifier.urihttp://hdl.handle.net/10722/178743-
dc.description.abstractThroughout spermatogenesis, inter-Sertoli tight junctions (TJs) that create the blood-testis barrier in the rat must be disassembled and reassembled to permit the timely passage of preleptotene spermatocytes from the basal to the adluminal compartment of the seminiferous epithelium. However, the mechanism(s) and the participating molecules that regulate this event are largely unknown. Although there is no in vitro model to study the event and regulation of inter-Sertoli TJ disassembly, primary cultures of Sertoli cells in vitro can be used to study junction assembly. In this study, we sought to investigate whether cytokines are involved in the inter-Sertoli TJ assembly in vitro. Sertoli cells isolated from 20-day-old rats were cultured at a density of 0.5-1.2 × 10 6 cells/cm 2 on Matrigel-coated dishes or bicameral units for 8-9 days. The steady-state messenger RNA levels of basic fibroblast growth factor (bFGF), transforming growth factor (TGF)-β2, and TGF-β3 at different time points were assessed by semiquantitative RT-PCR. In selected experiments, the assembly of inter-Sertoli TJs was monitored by transepithelial electrical resistance measurement. It was found that there was no change in the expression of basic fibroblast growth factor throughout the entire culture period. However, there was a 2-fold reduction in the expression of TGF-β2 and TGF-β3 at the time inter-Sertoli TJs were being assembled. On days 5-8, after the inter-Sertoli TJs had been assembled, the Sertoli cell steady-state messenger RNA levels of TGF-β2 and TGF-β3 increased by as much as 3- and 6-fold, respectively, when compared with Sertoli cells on days 1-3 when TJs were being assembled. Also, it was found that recombinant TGF-β3 added to Sertoli cells cultured in vitro at 1.2 × 10 6 cells/cm 2 on Matrigel-coated bicameral units perturbed the inter-Sertoli TJ permeability barrier dose-dependently. Moreover, the presence of TGF-β3 also inhibited the transient and/or basal expression of several TJ-associated proteins, which include occludin, zonula occludens-1, and claudin-11 when inter-Sertoli TJs were being assembled in vitro. These results suggest that TGF-β plays a crucial role in regulating the complicated biochemical events of junction assembly in the testis.en_US
dc.languageengen_US
dc.publisherThe Endocrine Society. The Journal's web site is located at http://endo.endojournals.orgen_US
dc.relation.ispartofEndocrinologyen_US
dc.rightsEndocrinology. Copyright © The Endocrine Society.-
dc.subject.meshAmino Acid Sequenceen_US
dc.subject.meshAnimalsen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshMaleen_US
dc.subject.meshMembrane Proteins - Chemistry - Genetics - Physiologyen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshNerve Tissue Proteinsen_US
dc.subject.meshPermeabilityen_US
dc.subject.meshPhosphoproteins - Physiologyen_US
dc.subject.meshRatsen_US
dc.subject.meshRats, Sprague-Dawleyen_US
dc.subject.meshRecombinant Proteins - Pharmacologyen_US
dc.subject.meshSertoli Cells - Metabolism - Ultrastructureen_US
dc.subject.meshTight Junctions - Drug Effects - Metabolismen_US
dc.subject.meshTransforming Growth Factor Beta - Pharmacologyen_US
dc.titleTransforming growth factor-β3 perturbs the inter-Sertoli tight junction permeability barrier in vitro possibly mediated via its effects on occludin, zonula occludens-1, and claudin-11en_US
dc.typeArticleen_US
dc.identifier.emailLui, WY: wylui@hku.hken_US
dc.identifier.emailLee, WM: hrszlwm@hku.hken_US
dc.identifier.authorityLui, WY=rp00756en_US
dc.identifier.authorityLee, WM=rp00728en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1210/en.142.5.1865en_US
dc.identifier.pmid11316752-
dc.identifier.scopuseid_2-s2.0-0035047036en_US
dc.identifier.hkuros56838-
dc.identifier.hkuros100782-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0035047036&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume142en_US
dc.identifier.issue5en_US
dc.identifier.spage1865en_US
dc.identifier.epage1877en_US
dc.identifier.isiWOS:000168434500025-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridLui, WY=35220192400en_US
dc.identifier.scopusauthoridLee, WM=24799156600en_US
dc.identifier.scopusauthoridCheng, CY=7404797787en_US

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