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

FITT MARCOS; BENITEZ, SERGIO; CASTRO, ANALÍA; J. RASMUSSEN; D. GALEANA; MUÑOZ, ESTELA

San Juan

Congreso; SECOND JOINT MEETING OF THE BIOLOGY SOCIETIES FROM ARGENTINA; 2011

In the post-genomic era, identifying gene products and revealingtheir roles represent an actual challenge. The pineal gland, throughits hormone melatonin, is a key effector and regulator of the circadian timing system. The differentiation factor NeuroD1 is thoughtto be a pineal phenotype determinant. Two NeuroD1 KO mice havebeen developed. Although new avenues have emerged, these models have disadvantages. Whereas the total KO mice die shortly after birth, the conditional KO mice lack NeuroD1 in both retina andpineal gland. Our group initiated controlled genetic manipulationin the rat pineal gland by optimizing the in utero intraventricularinjection and electroporation. The use of vectors carrying reportergenes driven by promiscuous promoters has revealed that the number of positive cells was highly dependent on the pineal mitoticactivity and time of injection. Highly skilled operators were neededto increase embryo survival rate. It is expected that pineal-selective gene targeting will help us to clarify unknown aspects of thepineal biology and its role in the general physiology. This procedure may also represent a cost-effective approach compared to themaintenance of transgenic mouse lines.