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CNDP1 knockout in zebrafish alters the amino acid metabolism, restrains weight gain, but does not protect from diabetic complications

Abstract

The gene CNDP1 was associated with the development of diabetic nephropathy. Its enzyme carnosinase 1 (CN1) primarily hydrolyzes the histidine-containing dipeptide carnosine but other organ and metabolic functions are mainly unknown. In our study we generated CNDP1 knockout zebrafish, which showed strongly decreased CN1 activity and increased intracellular carnosine levels. Vasculature and kidneys of CNDP1/ zebrafish were not affected, except for a transient glomerular alteration. Amino acid profiling showed a decrease of certain amino acids in CNDP1/ zebrafish, suggesting a specific function for CN1 in the amino acid metabolisms. Indeed, we identified a CN1 activity for Ala–His and Ser–His. Under diabetic conditions increased carnosine levels in CNDP1/ embryos could not protect from respective organ alterations. Although, weight gain through overfeeding was restrained by CNDP1 loss. Together, zebrafish exhibits CN1 functions, while CNDP1 knockout alters the amino acid metabolism, attenuates weight gain but cannot protect organs from diabetic complications.

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Abbreviations

CARNS1:

Carnosine synthase

CN1:

Carnosinase1 (enzyme)

CNDP1 :

Carnosinase1 (gene)

CN2:

Carnosinase2 (enzyme)

CNDP2 :

Carnosinase2 (gene)

DN:

Diabetic nephropathy

MG:

Methylglyoxal

pdx1:

Pancreas and duodenal homeobox 1

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Acknowledgements

The study was supported by grants from Deutsche Forschungsgemeinschaft (CRC 1118 and IRTG 1874/2 DIAMICOM). We thank the Metabolomics Core Technology Platform of the Excellence cluster “CellNetworks” (University of Heidelberg) and the Deutsche Forschungsgemeinschaft (Grant ZUK 40/2010-3009262) for support with UPLC-based metabolite quantification. We acknowledge the support of the Core Facility Live Cell Imaging (DFG INST 91027/10-1 FUGG).

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FS: conceived and designed the analysis; data collection; contribution of data or analysis tools; performed the analysis (mutant generation, fish work, phenotyping of the mutants); wrote the paper; critical revision of the paper; data analysis and interpretation; final approval of the version to publish. VP: conceived and designed the analysis; data collection; contribution of data or analysis tools; performed the analysis (CN1 activity, carnosine and anserine determinations, western blot); critical revision of the paper; data analysis and interpretation; final approval of the version to publish. CPS: conceived and designed the analysis; contribution of data or analysis tools; critical revision of the paper; data analysis and interpretation; final approval of the version to publish. GP: data collection; contribution of data or analysis tools; performed the analysis (metabolomic analysis); critical revision of the paper; final approval of the version to publish. MB: data collection; contribution of data or analysis tools; performed the analysis (metabolomic analysis); critical revision of the paper; final approval of the version to publish. XL: data collection; contribution of data or analysis tools; performed the analysis (fish work); critical revision of the paper; final approval of the version to publish. TW: data collection; contribution of data or analysis tools’; performed the analysis (western blot); critical revision of the paper; final approval of the version to publish. TP: data collection; contribution of data or analysis tools; performed the analysis (analysis and quantification of adult fish kidneys); critical revision of the paper; final approval of the version to publish. NV: data collection; contribution of data or analysis tools; critical revision of the paper; final approval of the version to publish. JM: data collection; contribution of data or analysis tools; performed the analysis (methylglyoxal and glucose determintation); critical revision of the paper; final approval of the version to publish. TF: data collection; contribution of data or analysis tools; performed the analysis (methylglyoxal and glucose determintation); critical revision of the paper; final approval of the version to publish. PPN: conceived and designed the analysis; data collection; contribution of data or analysis tools; critical revision of the paper; final approval of the version to publish. JK: conceived and designed the analysis; contribution of data or analysis tools; critical revision of the paper; data analysis and interpretation; final approval of the version to publish.

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Correspondence to Jens Kroll.

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Schmöhl, F., Peters, V., Schmitt, C.P. et al. CNDP1 knockout in zebrafish alters the amino acid metabolism, restrains weight gain, but does not protect from diabetic complications. Cell. Mol. Life Sci. 76, 4551–4568 (2019). https://doi.org/10.1007/s00018-019-03127-z

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Keywords

  • Amino acids
  • Carnosinase1
  • Carnosine
  • CRISPR/Cas
  • Diabetes
  • Diabetic nephropathy
  • Metabolism
  • Mutagenesis
  • Zebrafish