Immunohistochemical Detection of Vasa Antigen and Apoptosis-Related DNA Fragmentation in Ovaries of Sheep Fetuses Prenatally Exposed to Vitamin D Deficiency

Abstract

The primordial germ cells (PGCs) in female animals are comprised of diplotene oocytes arrested in the first meiotic prophase. Expression of Vasa is one of the key factors required for subsequent resumption of development and recruitment of PGCs into the growing follicle class. Since vitamin D regulates recruitment of PGCs and developmental competence of ovarian follicles, this study was designed to investigate the expression of Vasa and rate of apoptosis in foetal ovaries prenatally restricted from dietary vitamin D. Nineteen sexually mature Welsh mountain ewes were randomly assigned to vitamin D deficient (VDD) and vitamin D control (VDC) diets from 17d before mating, up to 125d of gestation, when fetal ovaries were collected and fixed in formalin for immunohistochemistry and TUNEL assay. VDD ovaries had fewer healthy oocytes that could stain positive for Vasa as well as a lower integrated density value for DAB staining intensity. Conversely, TUNNEL staining in VDD animals showed a higher integrated density value and percentage of affected area (P<0.05). The present findings indicate that Vasa expression is decreased, while the rate of apoptosis increased in VDD fetal ovaries, and this may adversely affect resumption of growth and development of PGCs reserve.

Keywords

• apoptosis
• ovary
• sheep
• vasa
• vitamin d deficiency