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Article: Interactions of proteases and protease inhibitors in sertoli-germ cell cocultures preceding the formation of specialized sertoli-germ cell junctions In Vitro

TitleInteractions of proteases and protease inhibitors in sertoli-germ cell cocultures preceding the formation of specialized sertoli-germ cell junctions In Vitro
Authors
KeywordsCathepsin D
Cathepsin L
Cathepsin S
Cell migration
Intercellular junction
Testis
Issue Date1997
PublisherAmerican Society of Andrology. The Journal's web site is located at http://www.andrologyjournal.org
Citation
Journal Of Andrology, 1997, v. 18 n. 6, p. 612-622 How to Cite?
AbstractThe biochemical mechanism(s) by which germ cells can form specialized junctions with Sertoli cells in the seminiferous epithelium at various stages of the spermatogenic cycle is unknown. This study sought to examine the biochemical changes that are involved when germ cells are cocultured with Sertoli cells in vitro preceding the establishment of specialized Sertoli- germ cell junctions. While isolated germ cells were allowed to attach to Sertoli cells, media from both the apical and basal compartments of bicameral units were collected to assess serine and cysteine protease activity. The expression of selected serine and cysteine proteases and their corresponding inhibitors in these Sertoli-germ cell cocultures was also examined by RT- PCR. Using an [125I]-collagen film assay, a transient but significant increase in serine protease activity was noted in both the apical and basal compartments when germ cells began to settle onto the Sertoli cell monolayer preceding the formation of intercellular junctions. A specific tryptase (RNK- Tryp 2, a serine protease formerly cloned from a rat granular lymphocyte leukemia cell line, RNK-16, cDNA expression library) was shown to be expressed exclusively by Sertoli cells and not germ cells. Furthermore, Sertoli cell tryptase expression as well as urokinase plasminogen activator (u-PA, also a serine protease) increased significantly when germ cells were adhering to Sertoli cells. The decline in total serine protease activity when Sertoli-germ cell junctions were being formed was accompanied by a concomitant increase in α2-macroglobulin (α2-MG, a nonspecific protease inhibitor) expression. No significant changes in cysteine protease activity in either the apical or basal compartment were noted. However, there was a transient but significant increase in cathepsin L expression when germ cells were adhering to Sertoli cells preceding cell junction formation. The subsequent reduction in cathepsin L expression after this transient increase was accompanied by a concomitant increase in cystatin C expression. These results suggest that proteases and their corresponding inhibitors are working synergistically and are likely to be involved in the adherence of germ cells to Sertoli cells and the subsequent formation of intercellular junctions.
Persistent Identifierhttp://hdl.handle.net/10722/49431
ISSN
2014 Impact Factor: 2.473
2015 SCImago Journal Rankings: 0.867
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorMruk, Den_HK
dc.contributor.authorZhu, LJen_HK
dc.contributor.authorSilvestrini, Ben_HK
dc.contributor.authorLee, WMen_HK
dc.contributor.authorCheng, CYen_HK
dc.date.accessioned2008-06-12T06:42:30Z-
dc.date.available2008-06-12T06:42:30Z-
dc.date.issued1997en_HK
dc.identifier.citationJournal Of Andrology, 1997, v. 18 n. 6, p. 612-622en_HK
dc.identifier.issn0196-3635en_HK
dc.identifier.urihttp://hdl.handle.net/10722/49431-
dc.description.abstractThe biochemical mechanism(s) by which germ cells can form specialized junctions with Sertoli cells in the seminiferous epithelium at various stages of the spermatogenic cycle is unknown. This study sought to examine the biochemical changes that are involved when germ cells are cocultured with Sertoli cells in vitro preceding the establishment of specialized Sertoli- germ cell junctions. While isolated germ cells were allowed to attach to Sertoli cells, media from both the apical and basal compartments of bicameral units were collected to assess serine and cysteine protease activity. The expression of selected serine and cysteine proteases and their corresponding inhibitors in these Sertoli-germ cell cocultures was also examined by RT- PCR. Using an [125I]-collagen film assay, a transient but significant increase in serine protease activity was noted in both the apical and basal compartments when germ cells began to settle onto the Sertoli cell monolayer preceding the formation of intercellular junctions. A specific tryptase (RNK- Tryp 2, a serine protease formerly cloned from a rat granular lymphocyte leukemia cell line, RNK-16, cDNA expression library) was shown to be expressed exclusively by Sertoli cells and not germ cells. Furthermore, Sertoli cell tryptase expression as well as urokinase plasminogen activator (u-PA, also a serine protease) increased significantly when germ cells were adhering to Sertoli cells. The decline in total serine protease activity when Sertoli-germ cell junctions were being formed was accompanied by a concomitant increase in α2-macroglobulin (α2-MG, a nonspecific protease inhibitor) expression. No significant changes in cysteine protease activity in either the apical or basal compartment were noted. However, there was a transient but significant increase in cathepsin L expression when germ cells were adhering to Sertoli cells preceding cell junction formation. The subsequent reduction in cathepsin L expression after this transient increase was accompanied by a concomitant increase in cystatin C expression. These results suggest that proteases and their corresponding inhibitors are working synergistically and are likely to be involved in the adherence of germ cells to Sertoli cells and the subsequent formation of intercellular junctions.en_HK
dc.format.extent418 bytes-
dc.format.mimetypetext/html-
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.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subjectCathepsin Den_HK
dc.subjectCathepsin Len_HK
dc.subjectCathepsin Sen_HK
dc.subjectCell migrationen_HK
dc.subjectIntercellular junctionen_HK
dc.subjectTestisen_HK
dc.titleInteractions of proteases and protease inhibitors in sertoli-germ cell cocultures preceding the formation of specialized sertoli-germ cell junctions In Vitroen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0196-3635&volume=18&issue=6&spage=612&epage=622&date=1997&atitle=Interactions+of+proteases+and+protease+inhibitors+in+Sertoli-germ+cell+cocultures+preceding+the+formation+of+specialized+Sertoli-germ+cell+junctions+in+vitroen_HK
dc.identifier.emailLee, WM: hrszlwm@hku.hken_HK
dc.identifier.authorityLee, WM=rp00728en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.pmid9432134-
dc.identifier.scopuseid_2-s2.0-0031424610en_HK
dc.identifier.hkuros22391-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0031424610&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume18en_HK
dc.identifier.issue6en_HK
dc.identifier.spage612en_HK
dc.identifier.epage622en_HK
dc.identifier.isiWOS:A1997YL44500007-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridMruk, D=6701823934en_HK
dc.identifier.scopusauthoridZhu, LJ=7404201524en_HK
dc.identifier.scopusauthoridSilvestrini, B=7006825900en_HK
dc.identifier.scopusauthoridLee, WM=24799156600en_HK
dc.identifier.scopusauthoridCheng, CY=7404797787en_HK

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