Effects of ethanol and nicotine co-administration on follicular atresia and placental histo-morphology in the first-generation mice pups during intrauterine development and lactation periods

Elahe Musanejad; Tahereh Haghpanah; Vida Mirzaie; Massood Ezzatabadipour

doi:10.1016/j.toxrep.2021.03.033

Effects of ethanol and nicotine co-administration on follicular atresia and placental histo-morphology in the first-generation mice pups during intrauterine development and lactation periods

Toxicol Rep. 2021 Apr 2:8:793-803. doi: 10.1016/j.toxrep.2021.03.033. eCollection 2021.

Authors

Elahe Musanejad¹, Tahereh Haghpanah¹, Vida Mirzaie¹, Massood Ezzatabadipour²

Affiliations

¹ Anatomical Sciences Department, School of Medicine, Kerman University of Medical Sciences, 76169-14115, Kerman, Iran.
² Neuroscience Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran.

Abstract

This study is evaluating the effects of ethanol and nicotine exposure during pregnancy and lactation on placenta histology and follicular atresia in the first-generation (f1) mice pups. The experimental groups were 5 groups of NMRI pregnant mice, including: control, vehicle (received normal saline) ethanol (3 g/kg/day, 20 % v/v intraperitoneally), nicotine (1 mg/kg/day, subcutaneously), and ethanol plus nicotine which received both. Pregnant animals in each group were then divided into two groups, one group for examining the placenta that was treated for 18 days and the other group for the ovary of one-day-old (PND1) and fifty-six-day-old (PND56) female offspring who were treated for 42 days (during intrauterine development and lactation). After the autopsy procedure, histopathological and morphometrical observations were done. Data revealed that the exposed mice had a significant change in the placenta morphometry and histology as well as a marked increase in the number of ovarian TUNEL positive cells on postnatal days 1 and 56. Therefore, maternal exposure to alcohol and nicotine during developmental and lactation periods could lead to changes in the placenta properties as well as an increase in the apoptotic ovarian follicles in f1 mice pups.

Keywords: AMH, Anti-Müllerian hormone; ANOVA, analysis of variance; BAX, BCL2 Associated X; BMP, Bone morphogenetic protein; Bcl-2, B-cell lymphoma 2; CTL, control; Ca2+, calcium; DAPI, 4′,6-diamidino-2-phenylindole; DNA, Deoxyribonucleic acid; E2, Estradiol; ELISA, enzyme-linked immunosorbent assay; EtOH, ethanol; Ethanol; FSH, Follicle-stimulating hormone; First-generation mice; Follicular atresia; GD, gestation day; H&E, Hematoxylin and Eosin; HCL, Hydrogen Chloride; IUGR, intrauterine growth restriction; NMRI, Naval Medical Research Institute; Ni, nicotine; Nicotine; OFR, ovarian follicular reservoir; OS, oxidative stress; PBS, Phosphate-buffered saline; PND, Postnatal day; Placenta; ROS, reactive-oxygen-species; SEM, standard error of the mean; SPSS, statistical package for the social sciences; TUNEL, Terminal deoxynucleotidyl transferase dUTP nick end labeling; Veh, vehicle; WHO, World Health Organization; cat.no, catalogue number; i.p., intraperitoneally; nAChRs, nicotinic acetylcholine receptors; s.c., subcutaneously.