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Article: Rat testicular extracellular superoxide dismutase: Its purification, cellular distribution, and regulation

TitleRat testicular extracellular superoxide dismutase: Its purification, cellular distribution, and regulation
Authors
Issue Date1998
PublisherSociety for the Study of Reproduction. The Journal's web site is located at http://www.biolreprod.org/
Citation
Biology Of Reproduction, 1998, v. 59 n. 2, p. 298-308 How to Cite?
AbstractUsing multiple HPLC steps, we have identified and purified a 68-kDa polypeptide (as estimated by gel permeation HPLC) to apparent homogeneity, from primary Sertoli cell-enriched culture medium, that consisted of two monomers of 35 (α chain) and 33 kDa (β chain) on SDS-polyacrylamide gel running under reducing conditions. Partial N-terminal amino acid sequence analysis of these two monomers revealed sequences of NH2-DXGESGVDIADRI (SOD(EX)-α) and NH2-XXDTGESGVDLADXL (SOD(EX)-β), which are identical to rat extracellular superoxide dismutase (SOD(EX)) with the exceptions that SOD(EX)-α and SOD(EX)β are missing, respectively, four (Trp-Thr-Met-Ser) and two (Trp-Thr) amino acids from their N-termini, compared to rat SOD(EX), suggesting that the cleavage sites of the SOD(EX) gene in the testis are different from that of other organs. Studies by sequential use of reverse transcription and polymerase chain reaction (PCR) using two SOD(EX) primers have demonstrated the expression of SOD(EX) in the heart, brain, lung, kidney, epididymis, testis, Sertoli, and germ cells, with low expression in the liver and ovary and no expression in the uterus, spleen, or thymus. Nucleotide sequence analysis of this 447-base pair PCR product from Sertoli cells revealed that its sequence is equivalent to the sequence of previously published rat SOD(EX). During testicular maturation, the SOD(EX) steady- state mRNA level increased significantly from 20 to 60 days of age and then declined at 90 days of age. Such an increase in the testicular SOD(EX) expression during maturation is not likely a result of an up-regulation by germ cells, since germ cells isolated from either 20- or 60-day-old rats when cocultured with Sertoli cells failed to elicit an increase in SOD(EX) expression in the cocultures. Using primary Sertoli cell cultures in vitro, it was found that Sertoli cell SOD(EX) expression was stimulated by interleukin-1α but not by either interferon-γ or basic fibroblast growth factor. These results illustrate that Sertoli cells as well as germ cells synthesize and/or secrete a testicular variant of SOD(EX) that may provide essential clues to understanding superoxide radical-mediated damage in the gonad.
Persistent Identifierhttp://hdl.handle.net/10722/49354
ISSN
2015 Impact Factor: 3.471
2015 SCImago Journal Rankings: 1.646
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorMruk, Den_HK
dc.contributor.authorCheng, CHen_HK
dc.contributor.authorCheng, YHen_HK
dc.contributor.authorMo, MYen_HK
dc.contributor.authorGrima, Jen_HK
dc.contributor.authorSilvestrini, Ben_HK
dc.contributor.authorLee, WMen_HK
dc.contributor.authorCheng, CYen_HK
dc.date.accessioned2008-06-12T06:40:12Z-
dc.date.available2008-06-12T06:40:12Z-
dc.date.issued1998en_HK
dc.identifier.citationBiology Of Reproduction, 1998, v. 59 n. 2, p. 298-308en_HK
dc.identifier.issn0006-3363en_HK
dc.identifier.urihttp://hdl.handle.net/10722/49354-
dc.description.abstractUsing multiple HPLC steps, we have identified and purified a 68-kDa polypeptide (as estimated by gel permeation HPLC) to apparent homogeneity, from primary Sertoli cell-enriched culture medium, that consisted of two monomers of 35 (α chain) and 33 kDa (β chain) on SDS-polyacrylamide gel running under reducing conditions. Partial N-terminal amino acid sequence analysis of these two monomers revealed sequences of NH2-DXGESGVDIADRI (SOD(EX)-α) and NH2-XXDTGESGVDLADXL (SOD(EX)-β), which are identical to rat extracellular superoxide dismutase (SOD(EX)) with the exceptions that SOD(EX)-α and SOD(EX)β are missing, respectively, four (Trp-Thr-Met-Ser) and two (Trp-Thr) amino acids from their N-termini, compared to rat SOD(EX), suggesting that the cleavage sites of the SOD(EX) gene in the testis are different from that of other organs. Studies by sequential use of reverse transcription and polymerase chain reaction (PCR) using two SOD(EX) primers have demonstrated the expression of SOD(EX) in the heart, brain, lung, kidney, epididymis, testis, Sertoli, and germ cells, with low expression in the liver and ovary and no expression in the uterus, spleen, or thymus. Nucleotide sequence analysis of this 447-base pair PCR product from Sertoli cells revealed that its sequence is equivalent to the sequence of previously published rat SOD(EX). During testicular maturation, the SOD(EX) steady- state mRNA level increased significantly from 20 to 60 days of age and then declined at 90 days of age. Such an increase in the testicular SOD(EX) expression during maturation is not likely a result of an up-regulation by germ cells, since germ cells isolated from either 20- or 60-day-old rats when cocultured with Sertoli cells failed to elicit an increase in SOD(EX) expression in the cocultures. Using primary Sertoli cell cultures in vitro, it was found that Sertoli cell SOD(EX) expression was stimulated by interleukin-1α but not by either interferon-γ or basic fibroblast growth factor. These results illustrate that Sertoli cells as well as germ cells synthesize and/or secrete a testicular variant of SOD(EX) that may provide essential clues to understanding superoxide radical-mediated damage in the gonad.en_HK
dc.format.extent418 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherSociety for the Study of Reproduction. The Journal's web site is located at http://www.biolreprod.org/en_HK
dc.relation.ispartofBiology of Reproductionen_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subject.meshTestis - cytology - enzymologyen_HK
dc.subject.meshChromatography, Gelen_HK
dc.subject.meshChromatography, High Pressure Liquiden_HK
dc.subject.meshCoculture Techniquesen_HK
dc.subject.meshElectrophoresis, Polyacrylamide Gelen_HK
dc.titleRat testicular extracellular superoxide dismutase: Its purification, cellular distribution, and regulationen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0006-3363&volume=59&issue=2&spage=298&epage=308&date=1998&atitle=Rat+testicular+extracellular+superoxide+dismutase:+its+purification,+cellular+distribution,+and+regulationen_HK
dc.identifier.emailLee, WM: hrszlwm@hku.hken_HK
dc.identifier.authorityLee, WM=rp00728en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.1095/biolreprod59.2.298en_HK
dc.identifier.pmid9687299-
dc.identifier.scopuseid_2-s2.0-0031879467en_HK
dc.identifier.hkuros34153-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0031879467&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume59en_HK
dc.identifier.issue2en_HK
dc.identifier.spage298en_HK
dc.identifier.epage308en_HK
dc.identifier.isiWOS:000075082700012-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridMruk, D=6701823934en_HK
dc.identifier.scopusauthoridCheng, CH=8630373700en_HK
dc.identifier.scopusauthoridCheng, YH=37093704600en_HK
dc.identifier.scopusauthoridMo, MY=7006857340en_HK
dc.identifier.scopusauthoridGrima, J=7003383792en_HK
dc.identifier.scopusauthoridSilvestrini, B=7006825900en_HK
dc.identifier.scopusauthoridLee, WM=24799156600en_HK
dc.identifier.scopusauthoridCheng, CY=7404797787en_HK

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