This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Add to My Marked Citations Citing Articles Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Tay, T. W. Articles by Kurohmaru, M. Search for Related Content PubMed PubMed Citation Articles by Tay, T. W. Articles by Kurohmaru, M. Social Bookmarking                         What's this?

Disappearance of Vimentin in Sertoli Cells: A Mono(2-ethylhexyl) Phthalate Effect

Department of Veterinary Anatomy, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan

Maki Ishii

Department of Developmental Neuroscience, Tokyo Metropolitan Institute for Neuroscience, Tokyo, Japan

Naoki Tsunekawa
Yoshiakira Kanai
Masamichi Kurohmaru

Department of Veterinary Anatomy, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan

Correspondence: Address correpondence to Masamichi Kurohmaru, Department of Veterinary Anatomy, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, 113-8657 Tokyo, Japan. E-mail:amkuroh{at}mail.ecc.u-tokyo.ac.jp

The effects of mono(2-ethylhexyl) phthalate (MEHP) on 21-day-old C57Bl/6N mice and their Sertoli cell cultures were studied. Mice were given a single dose of 800 mg/kg MEHP by oral gavage and sacrificed 24 h later. At the same time, testes were harvested from another batch of mice for Sertoli cell cultures. Cultures were subsequently exposed to 0, 1, and 100 nmol/ml MEHP for 0, 3, 6, 12, and 24 h. An antivimentin antibody was used to detect intermediate filament changes in Sertoli cells. Meanwhile, detection of preapoptotic signals and presence of apoptotic cells were done using annexin V–FITC (fluorescein isothiocyanate) and TUNEL (deoxynucleotidyltransferase-mediated dUTP nick end labeling) analyses, respectively. In vivo results showed a correlation between the increase in TUNEL-positive cells and the vimentin disruption in treated mice. Toluidine blue staining of the Sertoli cell cultures showed the increased number and size of vacuoles in Sertoli cell cytoplasm. Vimentin immunohistochemistry showed gradual disappearance of vimentin in Sertoli cell cultures as time and dose increased. Some Sertoli cells were found to be annexin V–FITC positive, but no TUNEL-positive cells were found. Taken together, these results show that the appearance of vacuoles and the vimentin disappearance caused by MEHP in the Sertoli cells are related with each other and can be observed in relation to time. This can be used as an indicator of the loss of mechanical support for spermatogenic cells, which in the end causes apoptosis of spermatogenic cells.

Key Words: Annexin V-FITC • Apoptosis • Mono(2-ethylhexyl) Phthalate • Testis • Vimentin

International Journal of Toxicology, Vol. 26, No. 4, 289-296 (2007)
DOI: 10.1080/00207450701470757


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?