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Article: Changes in the expression of junctional and nonjunctional complex component genes when inter-Sertoli tight junctions are formed in vitro

TitleChanges in the expression of junctional and nonjunctional complex component genes when inter-Sertoli tight junctions are formed in vitro
Authors
KeywordsIntercellular junction
Protease inhibitors
Proteases
Sertoli cells
Testis
Zonula occludens-1
Issue Date2000
PublisherAmerican Society of Andrology. The Journal's web site is located at http://www.andrologyjournal.org
Citation
Journal Of Andrology, 2000, v. 21 n. 2, p. 227-237 How to Cite?
AbstractThroughout spermatogenesis, germ cells move progressively from the basal to the adluminal compartment, which is accompanied by continual disassembly and reassembly of intercellular junctions suggesting germ cell movement is composed of intermittent phases of junction disassembly and reassembly. A study was performed to correlate the expression of junctional-complex components (such as zonula occludens-1 [ZO-1], a tight-junction component protein) and nonjunctional complex components (such as urokinase-type plasminogen activator [uPA], a serine protease; cathepsin L, a cysteine protease; α2-macroglobulin, a nonspecific protease inhibitor; and cystatin C, a cysteine protease inhibitor) at the time when inter-Sertoli tight junctions were established in vitro. This is an attempt to investigate whether the expression of nonjunctional component genes also correlates with the formation of inter-Sertoli tight junctions in vitro. This is part of an effort to understand the physiologic elements of germ cell movement in the epithelium. Sertoli cells cultured in vitro are known to undergo programmed cell death. To ensure that the changes in target gene expression were not the result of apoptosis, Sertoli cells were cultured in vitro at densities of 0.25, 0.75, and 3 x 106 cells/cm2 for up to 7 days on bicameral culture units coated with Matrigel (Collaborative Research) and were assessed by morphologic analysis and agarose gel electrophoresis. It was noted that many of the Sertoli cells cultured at 3 x 106 cells/cm2 underwent apoptosis by day 7, in contrast to cultures at 0.25 and 0.75 x 106 cells/cm2 illustrating the Sertoli cell number per unit of area may be an important parameter to be considered when studying Sertoli cell function in vitro. Also, it was shown that the expression of ZO-1 increased significantly between days 2 and 3 prior to the establishment of inter-Sertoli tight junctions assessed by transepithelial resistance measurement (TER), which illustrates that ZO-1 can be used as a marker to monitor this cellular event. More interestingly, there was also a transient increase in the expression of uPA and cathepsin L between days 2 and 3 at the time preceding the formation of tight junctions. In Sertoli cells cultured at low density (2 x 104 cells/cm2), when a confluent monolayer of cells could not form, there were no changes in the expression of either ZO-1, uPA, or cathepsin L throughout the 7-day culture period. These results show that the establishment of specialized junctions, such as tight junctions between Sertoli cells in vitro, may require the participation of both junctional and nonjunctional complex components.
Persistent Identifierhttp://hdl.handle.net/10722/84941
ISSN
2014 Impact Factor: 2.473
2015 SCImago Journal Rankings: 0.867
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorWong, CCSen_HK
dc.contributor.authorChung, SSWen_HK
dc.contributor.authorGrima, Jen_HK
dc.contributor.authorZhu, LJen_HK
dc.contributor.authorMruk, Den_HK
dc.contributor.authorLee, WMen_HK
dc.contributor.authorCheng, CYen_HK
dc.date.accessioned2010-09-06T08:58:54Z-
dc.date.available2010-09-06T08:58:54Z-
dc.date.issued2000en_HK
dc.identifier.citationJournal Of Andrology, 2000, v. 21 n. 2, p. 227-237en_HK
dc.identifier.issn0196-3635en_HK
dc.identifier.urihttp://hdl.handle.net/10722/84941-
dc.description.abstractThroughout spermatogenesis, germ cells move progressively from the basal to the adluminal compartment, which is accompanied by continual disassembly and reassembly of intercellular junctions suggesting germ cell movement is composed of intermittent phases of junction disassembly and reassembly. A study was performed to correlate the expression of junctional-complex components (such as zonula occludens-1 [ZO-1], a tight-junction component protein) and nonjunctional complex components (such as urokinase-type plasminogen activator [uPA], a serine protease; cathepsin L, a cysteine protease; α2-macroglobulin, a nonspecific protease inhibitor; and cystatin C, a cysteine protease inhibitor) at the time when inter-Sertoli tight junctions were established in vitro. This is an attempt to investigate whether the expression of nonjunctional component genes also correlates with the formation of inter-Sertoli tight junctions in vitro. This is part of an effort to understand the physiologic elements of germ cell movement in the epithelium. Sertoli cells cultured in vitro are known to undergo programmed cell death. To ensure that the changes in target gene expression were not the result of apoptosis, Sertoli cells were cultured in vitro at densities of 0.25, 0.75, and 3 x 106 cells/cm2 for up to 7 days on bicameral culture units coated with Matrigel (Collaborative Research) and were assessed by morphologic analysis and agarose gel electrophoresis. It was noted that many of the Sertoli cells cultured at 3 x 106 cells/cm2 underwent apoptosis by day 7, in contrast to cultures at 0.25 and 0.75 x 106 cells/cm2 illustrating the Sertoli cell number per unit of area may be an important parameter to be considered when studying Sertoli cell function in vitro. Also, it was shown that the expression of ZO-1 increased significantly between days 2 and 3 prior to the establishment of inter-Sertoli tight junctions assessed by transepithelial resistance measurement (TER), which illustrates that ZO-1 can be used as a marker to monitor this cellular event. More interestingly, there was also a transient increase in the expression of uPA and cathepsin L between days 2 and 3 at the time preceding the formation of tight junctions. In Sertoli cells cultured at low density (2 x 104 cells/cm2), when a confluent monolayer of cells could not form, there were no changes in the expression of either ZO-1, uPA, or cathepsin L throughout the 7-day culture period. These results show that the establishment of specialized junctions, such as tight junctions between Sertoli cells in vitro, may require the participation of both junctional and nonjunctional complex components.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.subjectIntercellular junctionen_HK
dc.subjectProtease inhibitorsen_HK
dc.subjectProteasesen_HK
dc.subjectSertoli cellsen_HK
dc.subjectTestisen_HK
dc.subjectZonula occludens-1en_HK
dc.titleChanges in the expression of junctional and nonjunctional complex component genes when inter-Sertoli tight junctions are formed in vitroen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0196-3635&volume=21&spage=227&epage=237&date=2000&atitle=Changes+in+the+Expression+of+Junctional+and+Nonjunctional+Complex+Component+Genes+When+Inter-Sertoli+Tight+Junctions+Are+Formed+In+Vitroen_HK
dc.identifier.emailLee, WM: hrszlwm@hku.hken_HK
dc.identifier.authorityLee, WM=rp00728en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.pmid10714817-
dc.identifier.scopuseid_2-s2.0-0033997927en_HK
dc.identifier.hkuros48611en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0033997927&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume21en_HK
dc.identifier.issue2en_HK
dc.identifier.spage227en_HK
dc.identifier.epage237en_HK
dc.identifier.isiWOS:000085540900008-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridWong, CCS=19037067100en_HK
dc.identifier.scopusauthoridChung, SSW=35104555300en_HK
dc.identifier.scopusauthoridGrima, J=7003383792en_HK
dc.identifier.scopusauthoridZhu, LJ=37048357500en_HK
dc.identifier.scopusauthoridMruk, D=6701823934en_HK
dc.identifier.scopusauthoridLee, WM=24799156600en_HK
dc.identifier.scopusauthoridCheng, CY=7404797787en_HK

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