Gerbes, Alexander L.

Nemer, M.

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Open Access LMU ( Ludwig-Maximilians-Univ. München)

Clin Investig (1993) 71:672 Molecular Medicine Rapid Communication Glinical Investigator © Springer-Verlag •993 Detection of C-type natriuretic peptide compared with brain and atrial natriuretic peptide transcripts in human heart by the polymerase chain reaction A.L. Gerbes 1' 2 M. Nemer  1 1 Laboratory of Molecular Genetics, Clinical Research Institute, Montreal 2 Medizinische Klinik II, Klinikum Groghadern, Universit/it Mtinchen C-type natriuretic peptide (CNP), initially isolated from porcine brain [3], is the most recently characterized member of the natriuretic peptide family. The human CNP gene consists of at least wo exons and an intron [4]. Whereas synthesis of atrial and brain atriuretic peptides (ANP and BNP) has been demonstrated mainly in the heart, but also in various other organs [2, 5], CNP mRNA in man has so far been found only in the central nervous system. We have investigated human cardiac tissues for CNP expression and compared the abun- dance of CNP transcripts with that of ANP and BNP. Atrial and ventricular tissues were obtained from a 76-year-old female patient with congestive heart failure at autopsy. Extrac- tion of total RNA, assessment ofRNA integrity, RNA quantifi- cation and cDNA synthesis by reverse transcriptase were per- formed as described in detail elsewhere [1]. The sense and anti- sense primers used for PCR amplification corresponded to se- quences in the first and second exon of the ANP, BNP and CNP genes, respectively. Thus, amplification of the cDNA and genomic DNA templates yielded fi'agments of 445 bp and 576 bp for ANP, 291 bp and 522 bp for BNP, and 358 bp and 802 bp for CNP. Various amounts (60-180 ng) of cDNA and 85 ng of human genomic DNA were coamplified, allowing for quantitation of the cDNA relative to total cDNA [1]. Amplifi- cation products were separated by agarose gel electrophoresis, transferred to nylon membranes (Hybond, Amersham) and hy- bridized to end-labelled oligonucleotides, corresponding to se- quences located between the two sequences used as PCR primers. Quantification was performed by image analysis densitometry of the hybridization bands. CNP transcripts of similar size were detected in brain, atrial and ventricular cDNA. Omission of cDNA served as negative control (Fig. 1). As summarized in Table 1, CNP transcripts were about three orders of magnitude l ss abundant than ANP and BNP transcripts in both atrial and ventricular tissue. We here present evidence for CNP expression in the human heart. Genomic + _ + + DNA cDNA Br At Vent ~802 bp - -  358  bp  Fig. 1. CNP transcripts in cDNA of brain (Br), atrial (At) and ventricular (Vent) tissue with or without added human genomic DNA Tab le  1. Natriuretic peptide cDNA abundance relative to geno- mic DNA cDNA/total cDNA Atrial Ventricular (% x 10- v) tissue tissue ANP 6770 •690 BNP 2840 •072 CNP 3.4 4.8 Since there is evidence for a pharmacological profile of CNP that is different from that of ANP and BNP [3], interest should focus on the potential role of CNP in cardiovascular physiology and pathophysiology. Acknowledgement. The work reported in this paper was Sup- ported by an ASCHE stipendium from the Deutsche Gesell- schaft ffir Verdauungs- und Stoffwechselkrankheiten to A.L.G. References 1. Dagnino L, Nemer M (1992) Increased transcripts for B-type natriuretic peptide in spontaneously h pertensive rats. Quan- titative polymerase chain reaction for atrial natriuretic and brain natriuretic peptide transcripts. Hypertension 20:690- 700 2. Gerbes AL, Dagnino L, Nguyen T, Nemer M (1992) Expres- sion of the brain natriuretic peptide gene in human tissues. Ninth International Congress of Endocrinology, Abstracts volume, p 154 3. Sudoh T, Minamino N, Kangawa K, Matsuo H (•990) C- type natriuretic peptide (CNP): a new member of the natriur- etic peptide family identified in porcine brain. Biochem Bio- phys Res Commun 1990; 168:863-870 4. Tawagari Y, Fuchimura K, Tanaka S, Minamino N, Kan- gawa K, Matsuo H (1991) Gene and precursor structures of human C-type natriuretic peptide. Biochem Biophys Res Commun 175 : 645-651 5. Vollmar AM (1990) Atrial natriuretic peptide in peripheral organs other than the heart. Klin Wochenschr 68 : 699-708 Received: April 7, 1993 Accepted:April 13, 1993 Dr. A.L. Gerbes Klinikum Grosshadern Marchionistrasse 15 D-81377 Miinchen, Germany 

Atrial natriuretic peptide in peripheral organs other than the heart.

Expression of the brain natriuretic peptide gene in human tissues.

Gene and precursor structures of human C-type natriuretic peptide.

Increased transcripts for B-type natriuretic peptide in spontaneously hypertensive rats. Quantitative polymerase chain reaction for atrial natriuretic and brain natriuretic peptide transcripts.

Matsuo H (•990) Ctype natriuretic peptide (CNP): a new member of the natriuretic peptide family identified in porcine brain. Biochem Biophys Res Commun

Detection of C-Type natriuretic peptide compared with brain and atrial natriuretic peptide transcripts in human heart by the polymerase chain reaction

A.L. Gerbest

M. Nemer

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Detection of C-type natriuretic peptide compared with brain and atrial natriuretic peptide transcripts in human heart by the polymerase chain reaction

https://epub.ub.uni-muenchen.de/5984/1/Gerbes_ALexander_5984.pdf

Detection of C-Type natriuretic peptide compared with brain and atrial natriuretic peptide transcripts in human heart by the polymerase chain reaction

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