Culturing rat whole ovary for UV filter benzophenone 3 treatment

Abstract

Here, we described a detailed protocol to establish a newborn rat whole ovary culture, able to study direct effects (independent of hypothalamic-pituitary-gonadal axis) of Endocrine Disrupting Chemicals (EDC), such as Benzophenone 3 (BP-3). This method is useful to understand changes in follicle formation, primordial to primary transition and expression of regulatory molecules linked to these processes as also provides an alternative to animal models. Basic protocols described here are the following:BASIC PROTOCOL 1: RAT OVARIAN SURGERYBASIC PROTOCOL 2: WHOLE ORGAN/OVARIAN CULTUREBASIC PROTOCOL 3: RNA ISOLATION AND QUANTITATIVE REAL TIME-PCRBASIC PROTOCOL 4: HISTOLOGICAL PROCESSING AND STAINING