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

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Label
  • mRNA expression in myocardial tissue of CNP, BNP and their 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 their 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
  • Biochimica Clinica (literal)
Rivista
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  • In: Biochimica Clinica, vol. 31 (5) pp. 1 -. BIOMEDIA srl, 2007. (literal)
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  • Background: It has been indirectly demonstrated that C-type natriuretic peptide (CNP) is produced in the myocardium in heart failure. Aim: to evaluate the expression of mRNA coding for CNP and its main receptor (NPR-B) in myocardial tissue by the use of RT-PCR. We also analysed BNP and NPR-A expression since cardiac tissue is the main site of synthesis for BNP. Methods: Total RNA was isolated from 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 the predominant receptor in the 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 chronic heart failure. (literal)
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  • Istituto di Fisiologia Clinica - CNR; Scuola Superiore Sant'Anna, Pisa (literal)
Titolo
  • mRNA expression in myocardial tissue of CNP, BNP and their main receptors NPR-B and NPR-A: a methodological assessment (literal)
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