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Article: Study on the formation of specialized inter-Sertoli cell junctions in vitro

TitleStudy on the formation of specialized inter-Sertoli cell junctions in vitro
Authors
Issue Date1999
PublisherJohn Wiley & Sons, Inc. The Journal's web site is located at http://www3.interscience.wiley.com/cgi-bin/jhome/31010
Citation
Journal Of Cellular Physiology, 1999, v. 181 n. 2, p. 258-272 How to Cite?
AbstractAn in vitro culture system using Sertoli cells was employed to assess the expression of component genes pertinent to occluding junctions (OJ) (such as zonula occludens-1, ZO-1), anchoring junctions (AJ) (such as N-cadherin and β-catenin), and communicating gap junctions (GJ) (such as connexin 33, Cx33) when they are being formed in vitro. Freshly isolated Sertoli cells from 20-day-old rats with a purity of greater than 90% were cultured either at low- (2.5 x 104 cells/cm2) or high-cell density (0.6 x 106 cells/cm2) on Matrigel-coated dishes for 7 days in vitro to allow the establishment of specialized junctions. In low cell density Sertoli cell cultures, specialized OJ such as tight junctions did not form during the entire culture period when assessed by the transepithelial electrical resistance (TER). In high cell density cultures, there was an increase in ZO-1 expression in days 1 to 3 preceding the establishment of tight junctions by day 4. When Sertoli cells were cultured at both cell densities, there was a transient increase in Sertoli cell N-cadherin expression, which peaked by days 4-5, suggesting the time course for the establishment of AJ may overlap with the OJ. A significant increase in the expression of Sertoli cell β-catenin was also detected by days 5-7 in the high but not low cell density cultures. The expression of Cx33 was also enhanced at days 4-5 in both high and low density cultures. These results suggest that OJ, AJ, and GJ are formed between Sertoli cells in high density cultures, whereas OJ cannot be formed in low density cultures. A full-length cDNA clone coding for rat testicular β- catenin was also isolated. The deduced amino acid sequence of rat β-catenin yielded a 781 amino acid polypeptide which displayed a 99.9% identity with the mouse homolog. Conditioned medium of germ cells induced a dose-dependent stimulation on Sertoli cell β-catenin expression, suggesting germ cells may affect the N-cadherin/β-catenin-mediated signal transduction pathway. In summary, this study illustrates several target genes can be used as molecular markers to monitor the inter-Sertoli junction formation. This system should be applicable to screen new male contraceptives in vitro targeted at the interference of junction formation by disrupting the timely expression of genes necessary for junction establishment and/or maintenance.
Persistent Identifierhttp://hdl.handle.net/10722/178659
ISSN
2015 Impact Factor: 4.155
2015 SCImago Journal Rankings: 1.842
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorChung, SSWen_US
dc.contributor.authorLee, WMen_US
dc.contributor.authorCheng, CYen_US
dc.date.accessioned2012-12-19T09:49:00Z-
dc.date.available2012-12-19T09:49:00Z-
dc.date.issued1999en_US
dc.identifier.citationJournal Of Cellular Physiology, 1999, v. 181 n. 2, p. 258-272en_US
dc.identifier.issn0021-9541en_US
dc.identifier.urihttp://hdl.handle.net/10722/178659-
dc.description.abstractAn in vitro culture system using Sertoli cells was employed to assess the expression of component genes pertinent to occluding junctions (OJ) (such as zonula occludens-1, ZO-1), anchoring junctions (AJ) (such as N-cadherin and β-catenin), and communicating gap junctions (GJ) (such as connexin 33, Cx33) when they are being formed in vitro. Freshly isolated Sertoli cells from 20-day-old rats with a purity of greater than 90% were cultured either at low- (2.5 x 104 cells/cm2) or high-cell density (0.6 x 106 cells/cm2) on Matrigel-coated dishes for 7 days in vitro to allow the establishment of specialized junctions. In low cell density Sertoli cell cultures, specialized OJ such as tight junctions did not form during the entire culture period when assessed by the transepithelial electrical resistance (TER). In high cell density cultures, there was an increase in ZO-1 expression in days 1 to 3 preceding the establishment of tight junctions by day 4. When Sertoli cells were cultured at both cell densities, there was a transient increase in Sertoli cell N-cadherin expression, which peaked by days 4-5, suggesting the time course for the establishment of AJ may overlap with the OJ. A significant increase in the expression of Sertoli cell β-catenin was also detected by days 5-7 in the high but not low cell density cultures. The expression of Cx33 was also enhanced at days 4-5 in both high and low density cultures. These results suggest that OJ, AJ, and GJ are formed between Sertoli cells in high density cultures, whereas OJ cannot be formed in low density cultures. A full-length cDNA clone coding for rat testicular β- catenin was also isolated. The deduced amino acid sequence of rat β-catenin yielded a 781 amino acid polypeptide which displayed a 99.9% identity with the mouse homolog. Conditioned medium of germ cells induced a dose-dependent stimulation on Sertoli cell β-catenin expression, suggesting germ cells may affect the N-cadherin/β-catenin-mediated signal transduction pathway. In summary, this study illustrates several target genes can be used as molecular markers to monitor the inter-Sertoli junction formation. This system should be applicable to screen new male contraceptives in vitro targeted at the interference of junction formation by disrupting the timely expression of genes necessary for junction establishment and/or maintenance.en_US
dc.languageengen_US
dc.publisherJohn Wiley & Sons, Inc. The Journal's web site is located at http://www3.interscience.wiley.com/cgi-bin/jhome/31010en_US
dc.relation.ispartofJournal of Cellular Physiologyen_US
dc.subject.meshAmino Acid Sequenceen_US
dc.subject.meshAnimalsen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCadherins - Geneticsen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshCloning, Molecularen_US
dc.subject.meshConnexins - Geneticsen_US
dc.subject.meshCytoskeletal Proteins - Biosynthesis - Chemistry - Geneticsen_US
dc.subject.meshGene Expression Regulationen_US
dc.subject.meshIntercellular Junctions - Physiologyen_US
dc.subject.meshMaleen_US
dc.subject.meshMembrane Proteins - Geneticsen_US
dc.subject.meshMiceen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshPhosphoproteins - Geneticsen_US
dc.subject.meshRatsen_US
dc.subject.meshRats, Sprague-Dawleyen_US
dc.subject.meshRecombinant Proteins - Biosynthesis - Chemistryen_US
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen_US
dc.subject.meshSequence Alignmenten_US
dc.subject.meshSequence Homology, Amino Aciden_US
dc.subject.meshSertoli Cells - Physiology - Ultrastructureen_US
dc.subject.meshSpermatozoa - Physiologyen_US
dc.subject.meshTrans-Activatorsen_US
dc.subject.meshTranscription, Geneticen_US
dc.subject.meshBeta Cateninen_US
dc.titleStudy on the formation of specialized inter-Sertoli cell junctions in vitroen_US
dc.typeArticleen_US
dc.identifier.emailLee, WM: hrszlwm@hku.hken_US
dc.identifier.authorityLee, WM=rp00728en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1002/(SICI)1097-4652(199911)181:2<258::AID-JCP8>3.0.CO;2-Qen_US
dc.identifier.pmid10497305-
dc.identifier.scopuseid_2-s2.0-0032876725en_US
dc.identifier.hkuros52466-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0032876725&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume181en_US
dc.identifier.issue2en_US
dc.identifier.spage258en_US
dc.identifier.epage272en_US
dc.identifier.isiWOS:000082935300008-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridChung, SSW=35104555300en_US
dc.identifier.scopusauthoridLee, WM=24799156600en_US
dc.identifier.scopusauthoridCheng, CY=7404797787en_US

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