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In Vivo Detection of Apoptotic Cells in the Testis Using Fluorescence Labeled Annexin V in a Mouse Model of Testicular Torsion

2006; Lippincott Williams & Wilkins; Volume: 176; Issue: 2 Linguagem: Inglês

10.1016/j.juro.2006.03.073

1527-3792

Sang‐Kuk Yang, Sharon Attipoe, Adam P. Klausner, Rong Tian, Dongfeng Pan, Tyvin A. Rich, Terry T. Turner, William D. Steers, Jeffrey J. Lysiak,

Reproductive System and Pregnancy

No AccessJournal of UrologyInvestigative urology1 Aug 2006In Vivo Detection of Apoptotic Cells in the Testis Using Fluorescence Labeled Annexin V in a Mouse Model of Testicular Torsionis companion ofThe Incidence of Bilateral Cryptorchidism is Increased and the Fertility Potential is Reduced in Sons Born to Mothers who Have Smoked During Pregnancy Sang-Kuk Yang, Sharon Attipoe, Adam P. Klausner, Rong Tian, Dongfeng Pan, Tyvin A. Rich, Terry T. Turner, William D. Steers, and Jeffrey J. Lysiak Sang-Kuk YangSang-Kuk Yang Department of Urology, Konkuk University School of Medicine, Chungju Hospital, Korea , Sharon AttipoeSharon Attipoe Department of Urology, University of Virginia, Charlottesville, Virginia , Adam P. KlausnerAdam P. Klausner Department of Urology, University of Virginia, Charlottesville, Virginia , Rong TianRong Tian Department of Radiology, University of Virginia, Charlottesville, Virginia , Dongfeng PanDongfeng Pan Department of Radiology, University of Virginia, Charlottesville, Virginia , Tyvin A. RichTyvin A. Rich Department of Radiation Oncology, University of Virginia, Charlottesville, Virginia , Terry T. TurnerTerry T. Turner Department of Urology, University of Virginia, Charlottesville, Virginia , William D. SteersWilliam D. Steers Department of Urology, University of Virginia, Charlottesville, Virginia , and Jeffrey J. LysiakJeffrey J. Lysiak Department of Urology, University of Virginia, Charlottesville, Virginia View All Author Informationhttps://doi.org/10.1016/j.juro.2006.03.073AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract Purpose: It is well established that experimental testicular torsion induces germ cell specific apoptosis. Annexin V (BD Pharmingen™) binds phosphatidylserine that becomes exposed on the cell membrane in apoptotic cells. In vivo detection of apoptotic cells with fluorescently labeled annexin V is an emerging technique that we evaluated for detecting apoptotic germ cells in a mouse model of testicular torsion. Materials and Methods: Annexin V labeled with an indocyanine fluorophore (bisfunctional succinimidyl ester of cyanine 5.5) (Amersham, Little Chalfont, United Kingdom) was injected intravenously in mice 18 hours after the repair of unilateral 720-degree testicular torsion for 2 hours. Serial fluorescence images were obtained 21, 24, 28 and 42 hours after torsion repair. Relative fluorophore localization was visualized in vivo using an optical small animal imaging system mounted with a filter in near infrared light. Average fluorescence intensity in torsed and sham testes was quantified in images of testes in situ exposed through an abdominal incision and in ex vivo testes. Results: A significant increase in fluorescence intensity was found in images of torsed vs sham operated testes. This was seen in ex vivo, exposed and in vivo testes (215%, 250% and 161%, respectively, p <0.05). Bisfunctional succinimidyl ester of cyanine 5.5 conjugated to dehydrogenase, a protein with a size similar to that of annexin V, was used to assess for capillary leakage. It was also more localized to the torsed testis relative to its contralateral sham control whether exposed or ex vivo (174% and 176%, respectively). Conclusions: To our knowledge this study demonstrates for the first time the possibility of in vivo near infrared fluorescence imaging of apoptotic germ cells after testicular torsion in mice. It shows important confounding factors that must be considered as this new imaging technique is developed for detecting apoptotic cells in vivo in testes or in any other organ. References 1 : Apoptosis: giving phosphatidylserine recognition an assist—with a twist. Curr Biol2003; 13: 655. Google Scholar 2 : In vivo analysis of phagocytosis of apoptotic cells by testicular Sertoli cells. Mol Reprod Dev2005; 71: 166. Google Scholar 3 : A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods1995; 184: 39. Google Scholar 4 : Role of apoptosis in reperfusion injury. Cardiovasc Res2004; 61: 414. 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Google Scholar © 2006 by American Urological AssociationFiguresReferencesRelatedDetailsRelated articlesJournal of Urology9 Nov 2018The Incidence of Bilateral Cryptorchidism is Increased and the Fertility Potential is Reduced in Sons Born to Mothers who Have Smoked During Pregnancy Volume 176Issue 2August 2006Page: 830-835 Advertisement Copyright & Permissions© 2006 by American Urological AssociationKeywordstorsionapoptosismicefluorescencetestisAcknowledgmentsDr. Bijoy Kundu, Small Animal Imaging Core, Department of Radiology, University of Virginia provided assistance.MetricsAuthor Information Sang-Kuk Yang Department of Urology, Konkuk University School of Medicine, Chungju Hospital, Korea More articles by this author Sharon Attipoe Department of Urology, University of Virginia, Charlottesville, Virginia More articles by this author Adam P. Klausner Department of Urology, University of Virginia, Charlottesville, Virginia More articles by this author Rong Tian Department of Radiology, University of Virginia, Charlottesville, Virginia More articles by this author Dongfeng Pan Department of Radiology, University of Virginia, Charlottesville, Virginia More articles by this author Tyvin A. Rich Department of Radiation Oncology, University of Virginia, Charlottesville, Virginia More articles by this author Terry T. Turner Department of Urology, University of Virginia, Charlottesville, Virginia More articles by this author William D. Steers Department of Urology, University of Virginia, Charlottesville, Virginia More articles by this author Jeffrey J. Lysiak Department of Urology, University of Virginia, Charlottesville, Virginia More articles by this author Expand All Advertisement PDF downloadLoading ...