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

Abstract

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.

Retraction: Quantitative morphometric analysis of vimentin filaments in Sertoli cells of rats after <i>in utero</i> DBP exposure

“…It has been reported that exposure of 3-week-old rats to di(2-ethlhexl)phthalate (DEHP) caused disruption and collapse of Vim microfilaments in Sertoli cells (Erkekoglu et al, 2012). Moreover, the volume of Vim microfilaments in Sertoli cells was reported to be reduced in young rats after exposure to other phthalates including DEHP, mono(2-ethylhexyl)phthalate (MEHP), and DBP (Richburg et al, 1999;Tay et al, 2007;Alam et al, 2010). Although one study concerning the alteration of Vim in Sertoli cells after in utero DBP exposure was reported, it was limited to five postnatal days (Kleymenova et al, 2005).…”

Apoptosis mediated by phosphatidylserine externalization in the elimination of aneuploid germ cells during human spermatogenesis

“…Furthermore, these cells are quiescent and remain in an irreversible G0 stage, and an absence of apoptotic markers has been described (Tay et al, 2007;Hassan et al, 2009;Stiblar-Martincic, 2009) possibly owing to a mechanism that protects them against programmed cell death (Johnson et al, 2008) and explaining our results.…”

Effects of di-iso-butyl phthalate on testes of prepubertal rats and mice

“…TUNEL-positive labeling was not found in Sertoli cells in rats 41−43) and mice [17][18][19]44) . In addition, both in vivo and in vitro experiments demonstrated that the Sertoli cell is the primary site of phthalate-induced testicular toxicity 45) .…”

“…This prompted us to focus our attention on primary effects of DiBP on Sertoli cells. Previous experiments demonstrated that MEHP causes the alternation of Sertoli cell cytoskeleton in mice and rats, particularly in distribution of vimentin filaments 19,41) . This study showed that vimentin filaments were partly disorganized or disappeared in the perinuclear and basal regions of Sertoli cells in the DiBP treated rat.…”