Validation of an alternative animal model for the development of a heterologous expression system of ion channels and membrane receptors

Abstract

The bullfrog (Lithobates catesbeianus) is originally from North American, but was introduced in Brazil in 1935, and nowadays it is commonly created in commercial ranches. Studies on morphological characterization of the stages of development of the female reproductive tract showed great similarity with the development of the Xenopus laevis frog. In both cases, it was possible to classify similar oocyte development. The oocytes of X. laevis have been used for decades as a heterologous expression model of membrane proteins in research laboratories around the world. However, this species of frog is not found in the Brazilian fauna and its importation faces bureaucratic difficulties involving different governmental organs, which practically makes it unfeasible, even for didactic and research purposes. Thus, in this research project we will evaluate the biotechnological potential of the use of L. catesbeianus oocytes as an alternative experimental model for the heterologous expression of membrane proteins. More specifically, we will perform electrophysiological analyzes of the present endogenous currents and a broad biophysical and pharmacological characterization of voltage-dependent ion channels and exogenous receptors that will be expressed in the oocyte plasma membrane. In addition, morphological analyzes of the plasma and yolk membrane will be made, as well as an evaluation of the distribution of ion channels and receptors along the membrane. (AU)