The use of laser microdissection coupled with semiquantitave PCR to study cell-specific gene expression during European sea bass (Dicentrarchus labrax) spermatogenesis

Abstract

The European sea bass (Dicentrarchus labrax), a gonochoristic marine teleost, is the subject of both basic and applied research. A major disadvantage of sea bass culture is the presence of precocious males that mature at the first year of life. One of the key points for understanding the molecular basis of male precocity is the quantification of gene expression in specific cell populations. We optimized a newly developed technique, Laser Microdissection Pressure Catapulting, coupled with semiquantitative PCR that allows determining the precise expression pattern of genes in specific cell populations throughout testis development. We quantified the expression pattern of several genes during the germ cell line progression from spermatogonia to spermatozoa that were already cloned in the sea bass. The studied genes were: androgen receptor (AR), P450 gonadal aromatase, vasa, and oestrogen receptors (ERα, ERβ1 and ERβ2). Knowledge of the specific expression levels of these genes will be useful to investigate their role in male spermatogenesis and their possible implications in the phenomenon of male precocity in sea bass aquaculture.