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Revista Científica
versión impresa ISSN 0798-2259
Resumen
RUIZ E, Ana Z y KITTOK, Roger. Dna amplification of the n-methyl-d-aspartate receptor subunit-1 (nmdar-1) in sheep hypothalamus by rt-pcr. Rev. Cient. (Maracaibo) [online]. 2006, vol.16, n.5, pp.496-505. ISSN 0798-2259.
The N-methyl-D-aspartate receptor (NMDAR), one of the excitatory amino acid receptors, has been implicated in the control of luteinizing hormone (LH) secretion through facilitation of luteinizing hormone releasing-hormone (LHRH). Interestingly, the ability of NMDAR to stimulate LHRH release is significantly compromised in castrated animals. The present study had three aims. Firstly, to standardize the optimal conditions used to amplify the DNA for NMDAR subtype-1 (NMDAR-1) in the sheep. Second, to determine whether a difference exists in the distribution of DNA for NMDAR-1 in different hypothalamic regions in intact male sheep. Finally, to determine if testicular steroid hormones are involved in NMDA receptor expression in the hypothalamus. The DNA of NMDAR-1 was amplified by RT-PCR in four quadrants of the hypothalamus (dorsal-rostral, ventral-rostral, dorsal-caudal, and ventral-caudal quadrant) of intact and castrated male sheep. The optimal conditions to amplify the DNA of NMDAR-1 were 1.5 mM MgCl2, 100 nM of primers and 59°C annealing temperature. In experiment 1, there was no effect of quadrant on the DNA expression of NMDAR-1 in the hypothalamus of the intact male sheep. Experiment 2 revealed that castration reduced (P < 0.05) the expression of DNA for NMDAR compared to that in intact males. Taken together, these data suggest that NMDAR-1 is widely expressed in the hypothalamus of male sheep. Furthermore these experiments suggest that testicular steroid hormones are involved in the regulation of NMDAR-1 expression at the DNA level, which allows understanding of its involvement in the modulation of different activities including reproduction
Palabras clave : Receptor NMDA; ovino; ADN patrón; RT-PCR; hormonas testiculares; NMDA receptor; sheep; template DNA; RT-PCR; testicular steroid hormones.
