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Article: Role of Tissue Inhibitor of Metalloproteases-1 in Junction Dynamics in the Testis

TitleRole of Tissue Inhibitor of Metalloproteases-1 in Junction Dynamics in the Testis
Authors
KeywordsAdherens junction
Germ cell
Protease inhibitor
Sertoli cell
Tight junction
Issue Date2003
PublisherAmerican Society of Andrology. The Journal's web site is located at http://www.andrologyjournal.org
Citation
Journal Of Andrology, 2003, v. 24 n. 4, p. 510-523 How to Cite?
AbstractUsing multiple high-performance liquid chromatography steps, we have identified and purified a polypeptide to apparent homogeneity from primary Sertoli cell conditioned culture medium that consisted of 2 molecular variants of 31 and 29 kDa when electrophoresed on a sodium dodecyl sulfate-polyacrylamide gel run under reducing conditions. Partial N-terminal amino acid sequence analysis of these 2 proteins revealed a sequence of NH 2-IKMA-KMLKGFDAVGNATG, which is homologous to tissue inhibitor of metalloproteases-1 (TIMP-1). Studies by semiquantitative reverse transcription-polymerase chain reaction using a primer pair specific to rat TIMP-1 demonstrated that both Sertoli and germ cells express TIMP-1. During maturation, the steady-state TIMP-1 mRNA level in the testis increased significantly from 40 to 60 days of age, which suggests its role in the restructuring of the epithelium during spermiation. This increase in testicular TIMP-1 expression was apparently not due to the increase in germ cell number, because TIMP-1 expression decreased approximately fivefold in germ cells isolated from testes of aging rats. Using Sertoli cells cultured at low (0.05 × 106 cells/cm2) and high (0.5 × 10 6 cells/cm2) densities, it was found that TIMP-1 expression increased transiently but significantly during junction assembly. A similar induction of TIMP-1 mRNA was also detected in Sertoli-germ cell cocultures during germ cell adhesion onto Sertoli cells. More important, the inclusion of either α2-macroglobulin (a protease inhibitor produced by Sertoli cells) or aprotinin (a serine protease inhibitor) into an in vitro germ cell adhesion assay facilitated the attachment of fluorescently labeled germ cells onto the Sertoli cell epithelium when compared to control, which suggests that the assembly of adherens junctions may involve protease inhibitors.
Persistent Identifierhttp://hdl.handle.net/10722/84837
ISSN
2014 Impact Factor: 2.473
2015 SCImago Journal Rankings: 0.867
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorMruk, DDen_HK
dc.contributor.authorSiu, MKYen_HK
dc.contributor.authorConway, AMen_HK
dc.contributor.authorLee, NPYen_HK
dc.contributor.authorLau, ASNen_HK
dc.contributor.authorCheng, CYen_HK
dc.date.accessioned2010-09-06T08:57:42Z-
dc.date.available2010-09-06T08:57:42Z-
dc.date.issued2003en_HK
dc.identifier.citationJournal Of Andrology, 2003, v. 24 n. 4, p. 510-523en_HK
dc.identifier.issn0196-3635en_HK
dc.identifier.urihttp://hdl.handle.net/10722/84837-
dc.description.abstractUsing multiple high-performance liquid chromatography steps, we have identified and purified a polypeptide to apparent homogeneity from primary Sertoli cell conditioned culture medium that consisted of 2 molecular variants of 31 and 29 kDa when electrophoresed on a sodium dodecyl sulfate-polyacrylamide gel run under reducing conditions. Partial N-terminal amino acid sequence analysis of these 2 proteins revealed a sequence of NH 2-IKMA-KMLKGFDAVGNATG, which is homologous to tissue inhibitor of metalloproteases-1 (TIMP-1). Studies by semiquantitative reverse transcription-polymerase chain reaction using a primer pair specific to rat TIMP-1 demonstrated that both Sertoli and germ cells express TIMP-1. During maturation, the steady-state TIMP-1 mRNA level in the testis increased significantly from 40 to 60 days of age, which suggests its role in the restructuring of the epithelium during spermiation. This increase in testicular TIMP-1 expression was apparently not due to the increase in germ cell number, because TIMP-1 expression decreased approximately fivefold in germ cells isolated from testes of aging rats. Using Sertoli cells cultured at low (0.05 × 106 cells/cm2) and high (0.5 × 10 6 cells/cm2) densities, it was found that TIMP-1 expression increased transiently but significantly during junction assembly. A similar induction of TIMP-1 mRNA was also detected in Sertoli-germ cell cocultures during germ cell adhesion onto Sertoli cells. More important, the inclusion of either α2-macroglobulin (a protease inhibitor produced by Sertoli cells) or aprotinin (a serine protease inhibitor) into an in vitro germ cell adhesion assay facilitated the attachment of fluorescently labeled germ cells onto the Sertoli cell epithelium when compared to control, which suggests that the assembly of adherens junctions may involve protease inhibitors.en_HK
dc.languageengen_HK
dc.publisherAmerican Society of Andrology. The Journal's web site is located at http://www.andrologyjournal.orgen_HK
dc.relation.ispartofJournal of Andrologyen_HK
dc.subjectAdherens junctionen_HK
dc.subjectGerm cellen_HK
dc.subjectProtease inhibitoren_HK
dc.subjectSertoli cellen_HK
dc.subjectTight junctionen_HK
dc.titleRole of Tissue Inhibitor of Metalloproteases-1 in Junction Dynamics in the Testisen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0196-3635&volume=24&issue=4&spage=210&epage=523&date=2003&atitle=Role+of+tissue+inhibitor+of+metalloproteases-1+in+junction+dynamics+in+the+testis.en_HK
dc.identifier.emailSiu, MKY: mkysiu@hkucc.hku.hken_HK
dc.identifier.emailLee, NPY: nikkilee@hku.hken_HK
dc.identifier.authoritySiu, MKY=rp00275en_HK
dc.identifier.authorityLee, NPY=rp00263en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.pmid12826691-
dc.identifier.scopuseid_2-s2.0-1342319723en_HK
dc.identifier.hkuros132875en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-1342319723&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume24en_HK
dc.identifier.issue4en_HK
dc.identifier.spage510en_HK
dc.identifier.epage523en_HK
dc.identifier.isiWOS:000183920500010-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridMruk, DD=6701823934en_HK
dc.identifier.scopusauthoridSiu, MKY=24924018400en_HK
dc.identifier.scopusauthoridConway, AM=36862532100en_HK
dc.identifier.scopusauthoridLee, NPY=7402722690en_HK
dc.identifier.scopusauthoridLau, ASN=7202626263en_HK
dc.identifier.scopusauthoridCheng, CY=7404797787en_HK

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