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Steroidogenesis in the offspring of rats submitted to maternal protein restriction: evaluation of estrogen-responsive microRNAs / mRNAs and functional prostate studies

Grant number: 18/26120-6
Support type:Scholarships in Brazil - Master
Effective date (Start): September 01, 2019
Effective date (End): May 31, 2021
Field of knowledge:Biological Sciences - Morphology
Principal Investigator:Luis Antonio Justulin Junior
Grantee:Luiz Marcos Frediani Portela
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Exposure to adverse conditions at critical periods of development, such as gestation and lactation, can lead to irreversible morphofunctional changes in the fetus, and this phenomenon is known as Fetal Programming (FP). Model widely used in FP, maternal protein restriction (MPR), is responsible for low birth weight and development of systemic changes in adult life. Recent research from our group has shown that MPR leads to delayed prostate development in young rats, increased estrogen/androgen ratio, and increased incidence of prostatic lesions with aging. In this way, the objectives of this project will be: 1) to evaluate the biosynthesis profile of steroid hormones (estrogen and testosterone) in the prostate and tissues with steroidogenic activity (testis, liver and brain) post natal 21 (PND 21); 2) Associate this data with alteration of estrogen responsive microRNAs / mRNAs in the ventral prostate (VP). For this, Sprague Dawley rats born from mothers fed normal ration (17% protein, CTR group) or with hypoprotein ration (6% protein, GLLP group) during gestation and lactation will be used. The animals will be euthanized in PND21, and the VPs, testis, liver and brain will be collected and stored in liquid nitrogen. The gene expression of steroidogenic pathway enzymes will be evaluated by RT-qPCR; the hormones and intermediary metabolites will be dosed in the blood by ELISA; immunohistochemical analyzes in PV for androgenic (AR) and estrogenic receptors (ER± and ER²). Expression levels of miR33-5p (previously selected from sequencing data) will be validated on rat PV samples. This microRNA will also be the target of functional studies from the treatment of normal human prostatic cells of the PNT-2 line treated with estrogen. Assays with 33-5p microRNA inhibitory and mimetic molecules will also be employed. In this case, target genes of this microRNA will be selected for validation by RT-qPCR. In this case, target genes of this microRNA will be selected for validation by RT-qPCR. These studies will aid in the understanding of the mechanisms of estrogenic action on the prostate of rats submitted to RPM. This project falls within the current research line of the responsible (FAPESP 2017 / 01063-7).