Drastic Loss of Antral Follicles Due to Gene Expression Dysregulation Occurs on the First Day After Subcutaneous Ovarian Transplantation

Abstract

Ovarian cryopreservation is an alternative for the preservation of fertility, and the subcutaneous transplantation site is considered one of the most promising. Studies evaluating the follicular growth and its relationship with gene expression and vascular perfusion are essential for improving this technique and its clinical application. Thus, the aim of this study was to evaluate the effect of subcutaneous autotransplantation and vitrification on follicular growth and atresia and their relationship with vascular perfusion and gene expression. Therefore, female mice were ovariectomized, and the ovaries were divided in two experimental groups (1) vitrified (treatment, n = 97) and (2) not vitrified (control, n = 97) and subsequently were transplanted. Then grafts, from both groups, were recovered after 1, 12, or 23 days (D1, D12, D23) and subjected to follicular quantification, morphometry, and qPCR. Non-transplanted ovaries (D0) were also used. The estrous cycle and vascular perfusion were monitored throughout the experiment. On D9, 100% of the animals had reestablished their estrous cycles (p > 0.05). Blood perfusion at the transplant site was similar for both treatments (p > 0.05), with greater perfusion at the site of vitrified transplants only on D1 (p < 0.05). A drastic reduction in the number of antral follicles and an increased number of atretic follicles were observed on D1 (p < 0.0001), associated with upregulation of Casp3, Fshr, and Igf1r; and downregulation of Bax, Acvr1, Egfr, and Lhcgr (p < 0.05). Our findings indicate that the first day after subcutaneous transplantation is a critical period for follicular survival, with intense follicular atresia independent of Bax upregulation.

Keywords: Atresia; Fertility; Folliculogenesis; Ovary; Transplant; Vitrification.