Characterization of n-myc downstream regulated 3 (ndrg3) during Medaka development

ARIAS PADILLA, L.F.; FERNANDINO, J.I.

Okazaki

Conferencia; 22nd Japanese Medaka and Zebrafish Meeting (JMZM).; 2016

National Institute of Basic Biology

The molecular and cellular basis of the development of a functional and differentiated gonad (ovary or testis) starting from an unique primordium, corresponding to an exceptional and exclusive embryological event, in contrast to other organ rudiments that differentiate into only one type of organ. In the last decade have been identified many genes involved in different moments of the development of a gonad, but is necessary deepen the study of these to comprehend the molecular cascade during clue moments of development, as proliferation, apoptosis and angiogenesis. Particularly, in this study the principal focus has been characterized the gene n-myc downstream regulated 3 (ndrg3) during the development to determinate if this could be involved in a clue moment of gonadogenesis. Initially was analyzed the ndrg3 expression pattern during different stages of development using medaka (Oryzias latipes, himedaka strain) by whole-mount in situ hybridization (WISH). The ndrg3 transcript was found since 9 somite stage (stage 22), and during all of the rest of embryo development stages in different organs, principally in kidney; however the expression in juvenile medaka post hatching stages also will be analyzed to complete the pattern of expression in relation to the gonad development. On the other hand, was analyzed the partial putative regulatory region sequence of ndrg3 gene, for which was obtain in silico 1282 pb sequence upstream of the putative transcriptional initiation site and analyzed in different bioinformatic tools, which have reveled different transcription factor binding sites, like AR (Androgen receptor) and ER (Estrogen Receptor) responsive elements, which is a good baseline data for exploring the molecular basis of the regulation of ndrg3, thus the midterm plan is analyze the regulation by those sex steroids in-vitro and ex-vivo approach. In addition to these results, the principal plan is continue with the study of the ndrg3 function during gonadogenesis, with the analysis of the phenotype using a knockout approach.