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Updated: Mar 30, 2026
Rapid and Efficient Spatiotemporal Monitoring of Normal and Aberrant Cytosine Methylation within Intact Zebrafish Embryos
Published on: August 18, 2022
Temporal and organ-specific detection of cNMPs including cUMP in the zebrafish
Fanni Dittmar1, Salim Abdelilah-Seyfried2, Sarah K Tschirner3
1Institute of Pharmacology, Hannover Medical School, Carl-Neuberg-Str. 1, D-30625 Hannover, Germany.
Abstract:
The cyclic pyrimidine nucleotides cCMP and cUMP occur in mammalian cell lines. Recently, cCMP was also identified in mouse organs. Due to technical difficulties, it has not been possible to detect cUMP in organs or tissues yet. Here, we have generated a temporal profile of the occurrence of nucleoside 3',5'-cyclic monophosphates during different developmental stages of embryogenesis and in different organs of the adult zebrafish Danio rerio. Cyclic nucleotides were quantified by high performance liquid chromatography quadrupole tandem mass spectrometry. The identity of cCMP and cUMP in the zebrafish was confirmed by high performance liquid chromatography quadrupole time-of-flight mass spectrometry. We show for the first time that cUMP can be detected during embryogenesis and in adult organs of this vertebrate model system.
Insights
Cyclic pyrimidine nucleotides, including cyclic uridine monophosphate (cUMP), are now detectable in zebrafish during embryogenesis and in adult organs. This breakthrough overcomes previous technical challenges in detecting cUMP in tissues.
Area of Science:
- Biochemistry
- Developmental Biology
- Molecular Biology
Background:
- Cyclic pyrimidine nucleotides, cyclic cytidine monophosphate (cCMP) and cyclic uridine monophosphate (cUMP), are known to exist in mammalian cell lines.
- cCMP has been identified in mouse organs, but technical limitations have prevented the detection of cUMP in organs or tissues.
- Understanding the distribution and role of these cyclic nucleotides is crucial for comprehending cellular signaling pathways.
Purpose of the Study:
- To establish a temporal profile of nucleoside 3',5'-cyclic monophosphates during zebrafish embryogenesis and in adult organs.
- To overcome technical challenges and enable the detection of cUMP in a vertebrate model system.
- To investigate the occurrence of cCMP and cUMP in the zebrafish (Danio rerio) across different developmental stages and adult tissues.
Main Methods:
- Quantification of cyclic nucleotides using high-performance liquid chromatography quadrupole tandem mass spectrometry (HPLC-MS/MS).
- Confirmation of cCMP and cUMP identity in zebrafish using high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS).
- Analysis of cyclic nucleotide occurrence during various embryonic developmental stages and in different organs of adult zebrafish.
Main Results:
- The study successfully generated a temporal profile of cyclic pyrimidine nucleotide occurrence in zebrafish.
- For the first time, cUMP was detected during zebrafish embryogenesis.
- cUMP was also detected in various organs of adult zebrafish, overcoming previous detection limitations.
Conclusions:
- This research demonstrates the successful detection of cUMP in a vertebrate model system (zebrafish) during embryogenesis and in adult organs.
- The findings provide a foundation for future studies on the biological roles of cUMP in vertebrate development and physiology.
- The developed methodology enables the investigation of cyclic pyrimidine nucleotides in contexts previously limited by technical challenges.
