Hum. Reprod. Advance Access originally published online on November 25, 2005
Human Reproduction 2006 21(3):610-617; doi:10.1093/humrep/dei393
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Nicotine does not affect vascularization but inhibits growth of freely transplanted ovarian follicles by inducing granulosa cell apoptosis
1 Department of Experimental Surgery, University of Rostock, Schillingallee 70, 18055 Rostock, Germany and 2 Department of Clinical and Experimental Surgery, University of Saarland, 66421 Homburg/Saar, Germany
3 To whom correspondence should be addressed. E-mail: brigitte.vollmar{at}med.uni-rostock.de
BACKGROUND: There is clear support for an association between smoking and decreased female fecundity and fertility. Cigarette smoke appears to have adverse effects along a continuum of reproductive processes. We therefore studied the effect of nicotine on follicular growth and vascularization of freely transplanted ovarian follicles. METHODS: We used the skinfold chamber model in Syrian golden hamsters, which allows the in vivo microscopy of follicular grafts. Animals were treated daily with nicotine at doses mimicking low-rate and high-rate smokers (0.2 and 1.0 mg/kg body weight subcutaneously). Saline-treated animals served as controls. To further evaluate the effect of nicotine on angiogenesis, an in vitro aortic ring assay was used. RESULTS: The re-vascularisation rate of follicles was similar in nicotine-treated animals and controls. During the 7 days after transplantation, nicotine further caused a dose-dependent inhibition of follicular growth. In contrast, the vascularized area and microvessel density were not affected by the nicotine exposure. In vitro aortic ring assays confirmed that nicotine does not influence sprouting and microvessel formation. However, immunohistochemistry for cleaved caspase-3 revealed a large extent of granulosa cell apoptosis within transplanted follicles of high-dose nicotine-treated animals. CONCLUSIONS: Nicotine as one toxic component of cigarette smoke does not affect vascularization, but adversely influences follicular growth by an increase in apoptotic cell death. As follicular growth is a crucial step in normal ovulation and fertilization, nicotine-induced cell apoptosis may represent one of the mechanisms underlying the well-established link between smoking and fertility disorders.
Key words: aortic ring assay/apoptosis/growth/ovarian follicles/vascularization
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