Expression of mRNAs of AngII receptors by RT-PCR and Northern Blot in cerebellum

3. MANZUR, MJ, ARCE ME, MUÑOZ RV, GIL L ANDREA F, ALVAREZ SE, CIUFFO GM.

Iguazú, Misiones, República Argentina

Congreso; XL Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular- SAIB; 2004

EXPRESSION OF mRNAs OF ANG II RECEPTORS BY RT-PCR AND NORTHERN BLOT IN CEREBELLUM. Manzur María J, Arce María E, Muñoz, Rosana V, Gil Lorenzo, Andrea F, Alvarez Sergio E and Ciuffo Gladys M. Bioquímica Avanzada. Fac. Qca, Bioqca y Fcia. UNSL. Ejército de los Andes 950 (5700) San Luis.  E-mail: mjmanzu@unsl.edu.ar Angiotensin II (Ang II) exerts its physiological effects through binding to membrane receptors, classified by their pharmacological characteristics as AT1 and AT2 receptors. Ang II receptor expression is highly modulated during development suggesting a role of these receptors in growth and organogenesis. With the aim to explore the expression pattern of Ang II receptors, we subcloned  PCR fragments of  AT1 and AT2 receptors in the p-GEM T easy vector.   The identity of the amplification products and subcloned inserts was verified by RFLP. For pGem-AT1 we used Ssp I,  Pvu II, Dde I and Not I and for pGem-AT2 the restriction endonucleases used were EcoR I, Ssp I and Pvu II. The inserts were extracted from the vector with restriction enzymes and the band excised from the gel and purified. The subcloned fragment was labeled with 35S ATP by random priming and used as a probe for Northern blot analysis.  Rat cerebellum mRNAs of different ages were analyzed. The expression pattern was compared with RT-PCR analysis of the same samples. The highest expression of AT2 receptor was observed at the age of 15.