http://www.cnr.it/ontology/cnr/individuo/prodotto/ID109117

mRNA expression in myocardial tissue of CNP, BNP and its main receptors NPR-B and NPR-A: a methodological assessment (Abstract in rivista)

Type

Abstract in rivista (Classe)
Prodotto della ricerca (Classe)

Label

mRNA expression in myocardial tissue of CNP, BNP and its main receptors NPR-B and NPR-A: a methodological assessment (Abstract in rivista) (literal)

Anno

2007-01-01T00:00:00+01:00 (literal)

Alternative label

Del Ry S.; Cabiati M.; Colotti C.; Lionetti V.; Maltinti M.; Giannessi D. (2007)
mRNA expression in myocardial tissue of CNP, BNP and its main receptors NPR-B and NPR-A: a methodological assessment
(literal)

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Del Ry S.; Cabiati M.; Colotti C.; Lionetti V.; Maltinti M.; Giannessi D. (literal)

Convegno

Clinical Chemistry and Laboratory Medicine (literal)

Rivista

Clinical chemistry and laboratory medicine (Rivista)

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In: Clinical Chemistry and Laboratory Medicine, vol. 45 pp. w132 -. S1-S473 EUROMEDLAB Amsterdam 2007. Walter de Gruyter, 2007. (literal)

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Background: Recently C-type natriuretic peptide (CNP) was supposed to be produced directly in the myocardium. Aim: to investigate the expression of mRNA coding for CNP and its main receptor (NPR-B) in myocardial tissue by use of RT-PCR. Since cardiac tissue is the main site of synthesis for BNP we also analysed BNP and NPR-A expression. Methods: Total RNA was isolated from the cardiac tissue samples (BALB/c mice, n=5) by guanidinium thyocyanate-phenol-chloroform method. To check the sensitivity and linearity of the amplification, RT-PCR for each primer was performed in a range of different number of cycles, annealing temperatures and RNA concentrations. GAPDH mRNA expression was determinated as internal control. Results: The optimal reaction conditions chosen were: 0.2 microg of RNA; annealing time and temperature of 60 s at 59°, 61°, 69°, 63°, 57°C for GAPDH, BNP, CNP, NPR-B, NPR-A primers, respectively; 30 cycles for BNP, NPR-A and GAPDH, 40 for CNP and 35 for NPR-B; extension time of 72°C. Bands were analyzed by Quanty One Software. The proposed method allowed us to detect the expression of mRNA coding for CNP and NPR-B in myocardial tissue confirming the presence of this effector in the heart. These preliminary data appear to indicate that CNP mRNA expression is lower with respect to that of BNP (CNP/GAPDH=0.117 vs. BNP/GAPDH=0.247) and that NPR-B is predominant in heart (NPR-A/GAPDH=0.244±0.028; NPR-B/GAPDH=0.657±0.022; p=0.0008). Conclusions: the determination of cardiac mRNA expression of CNP and NPR-B could constitute an useful tool to better study the role of this natriuretic peptide in experimental animal models of cronic heart failure. (literal)

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