GENETIC MANIPULATION IN THE RAT PINEAL GLAND: IN UTERO INTRAVENTRICULAR INJECTION AND ELECTROPORATION

M. FITT; A.CASTRO; S. BENITEZ; J. RASMUSSEN; D. GALEANA; E. MUÑOZ

San Juan

Congreso; Segunda Reunión Conjunta de las Sociedades de Biología; 2011

Sociedades de Biología

GENETIC MANIPULATION IN THE RAT PINEAL GLAND: IN UTERO INTRAVENTRICULAR INJECTION AND ELECTROPORATION. Fitt M, Castro AE, Benitez SG, Rasmussen J, Galiana D and Muñoz EM. IHEM-CONICET, FCM-UNCuyo, Mendoza, Argentina. ANPCyT. E-mail: munoz.estela@fcm.uncu.edu.ar In the post-genomic era, identifying gene products and elucidating their roles represent a significant challenge. The pineal gland, through its hormone melatonin, is a key effector and regulator of the circadian timing system. The differentiation factor NeuroD1 is thought to be a pineal phenotype determinant. Two NeuroD1 KO mice have been developed. Although new avenues have emerged from their use, these models have disadvantages. Whereas the total KO mice die shortly after birth, there are conditional KO mice that lack NeuroD1 in both the retina and pineal gland. Our group initiated controlled genetic manipulation in the rat pineal gland through in utero intraventricular injection and electroporation. The use of vectors carrying reporter genes driven by promiscuous promoters has revealed that the number of positive cells was highly dependent on the pineal mitotic activity and time of injection. Highly skilled operators were needed to increase the embryo survival rate. It is expected that pineal-selective gene targeting will help us to clarify unknown aspects of the pineal biology. This procedure may also represent a cost-effective approach compared to the maintenance of transgenic mouse lines.