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  • Meeting abstract
  • Open Access

Zebrafish as model organism for cNMP research

  • 1Email author,
  • 2,
  • 3 and
  • 1
BMC Pharmacology and Toxicology201516 (Suppl 1) :A45

  • Published:


  • Cyclic Nucleotide
  • Zebrafish Embryo
  • Chromatography Tandem Mass Spectrometry
  • Liquid Chromatography Tandem Mass Spectrometry
  • Liquid Chromatography Tandem Mass


The zebrafish Danio rerio has become an important vertebrate model organism for a wide range of scientific research questions including development, genetics, and disease [1]. The zebrafish is particularly well suited for this research due to its small size, rapid development, short generation time, optical transparency of embryos and larvae as well as functional conservation of genes [1]. Furthermore, application of drugs is easy since zebrafish readily absorb compounds from their surrounding media [2].

The aim of our study was to determine the composition and to quantify the endogenous level of different cyclic nucleotides (cNMPs) in various developmental stages and organs of Danio rerio.


Wild-type AB zebrafish were mated; for tissue preparation, we harvested embryos at 24 hours post fertilization (hpf) and larvae at 5 days post fertilization (dpf). Organs (eyes, brain, heart, entrails, eggs and testes) from adult female and male wild-type AB zebrafish were dissected according to the guidance of Gupta and Mullins [3]. All samples were then prepared for measurement of cNMP concentrations via a highly sensitive and specific method, namely high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) [4].


We could detect all studied cNMPs (namely cAMP, cCMP, cGMP, cIMP and cUMP) with specific variations as outlined below. (1) cGMP and cAMP could be detected in tissue samples of both developmental stages and within all harvested organs. Remarkably, they are the only cyclic nucleotides detected in brain and entrails. The highest cGMP concentrations were present in eyes; the highest cAMP concentrations in larvae tissue. (2) cIMP was only detected in some organs with the highest concentration in heart. (3) cCMP could not be detected in embryos but in larvae and within several organs. Like cIMP, the highest cCMP concentrations were present in heart. (4) cUMP was detected in both developmental stages as well as in some organs. This cNMP was most abundant within the heart.


The zebrafish Danio rerio constitutes a good model organism not only for studies focused on development, genetics and disease but also in the field of cyclic nucleotides, especially related to the role of cNMPs in cardiovascular biology. We observed specific cNMP patterns in development and in different organs, which is in support of the hypothesis of a distinct cNMP signaling code [5].
Table 1

cNMP concentrations of zebrafish embryos, larvae and various organs.


cGMP[pmol/mg protein]

cIMP[pmol/mg protein]

cAMP[pmol/mg protein]

cCMP[pmol/mg protein]

cUMP[pmol/mg protein]

Embryos 24 hpf

0.282 ± 0.009

not detected

2.29 ± 0.37

not detected

0.414 ± 0.067

Larvae 5 dpf

1.99 ± 0.32

not detected

23.66 ± 3.97

0.328 ± 0.119

0.890 ± 0.422


2.69 ± 0.03

0.111 ± 0.000

2.73 ± 0.34

0.043 ± 0.004

0.087 ± 0.040


0.149 ± 0.073

not detected

10.02 ± 2.89

not detected

not detected


0.607 ± 0.124

0.452 ± 0.069

23.12 ± 17.40

0.504 ± 0.143

1.11 ± 0.10


0.292 ± 0.031

not detected

16.52 ± 2.30

not detected

not detected


0.002 ± 0.003

not detected

0.29 ± 0.02

0.004 ± 0.001

0.008 ± 0.001


0.802 ± 1.134

0.407 ± 0.037

12.15 ± 3.80

0.116 ± 0.164

0.617 ± 0.052

Future studies

Zebrafish embryos will be treated with the NO-independent sGC activator cinaciguat (BAY 58-2667) and the NO-synergistic sGC stimulator 3-(4-amino-5-cyclopropylpyrimidine-2-yl)-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine (BAY 41-2272), from one day post fertilization to five days post fertilization. cNMP concentrations and morphological changes will be determined afterwards. Another research area will be the elucidation of the controversial role of cIMP as second messenger [6, 7].

Authors’ Affiliations

Institute of Pharmacology, Hannover Medical School (MHH), Hannover, Germany
Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany
Research Core Unit Metabolomics, Institute of Pharmacology, Hannover Medical School (MHH), Hannover, Germany


  1. Lawrence C: The husbandry of zebrafish (Danio rerio): A review. Aquaculture. 2007, 269 (1-4): 1-20. 10.1016/j.aquaculture.2007.04.077.View ArticleGoogle Scholar
  2. Langheinrich U: Zebrafish: A new model on the pharmaceutical catwalk. BioEssays. 2003, 25 (9): 904-912. 10.1002/bies.10326.View ArticlePubMedGoogle Scholar
  3. Gupta T, Mullins MC: Dissection of organs from the adult zebrafish. J Vis Exp. 2010, 37Google Scholar
  4. Bähre H, Danker KY, Stasch JP, Kaever V, Seifert R: Nucleotidyl cyclase activity of soluble guanylyl cyclase in intact cells. Biochem Biophys Res Commun. 2014, 443 (4): 1195-1199. 10.1016/j.bbrc.2013.12.108.View ArticlePubMedGoogle Scholar
  5. Seifert R, Schneider EH, Bähre H: From canonical to non-canonical cyclic nucleotides as second messengers: Pharmacological implications. Pharmacol Ther. 2015, 148: 154-184.View ArticlePubMedGoogle Scholar
  6. Chen Z, Zhang X, Ying L, Dou D, Li Y, Bai Y, et al: cIMP synthesized by sGC as a mediator of hypoxic contraction of coronary arteries. AJP Hear Circ Physiol. 2014, 307 (3): H328-H336. 10.1152/ajpheart.00132.2014.View ArticleGoogle Scholar
  7. Seifert R: Is cIMP a second messenger with functions opposite to those of cGMP?. Naunyn Schmiedebergs Arch Pharmacol. 2014, 387 (9): 897-899. 10.1007/s00210-014-1013-1.View ArticlePubMedGoogle Scholar


© Dittmar et al. 2015

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