Decreased cADPR and increased NAD+ in the Cd38−/− mouse

doi:10.1016/j.bbrc.2006.05.100

Biochemical and Biophysical Research Communications

Volume 346, Issue 1, 21 July 2006, Pages 188-192

https://doi.org/10.1016/j.bbrc.2006.05.100 Get rights and content

Abstract

CD38 is a type II glycoprotein that catalyzes the formation of cyclic ADP-ribose (cADPR), an intracellular calcium signalling molecule, from nicotinamide adenine dinucleotide (NAD⁺). Using a modified version of the fluorimetric cycling assay for cADPR which reduces between-subject variability, we report significant decreases in brain and lung cADPR, which although similar to previously published values, showed much less individual variation. The reduced variation within each group suggests that the range of cADPR is narrower than previously thought, and that the regulatory mechanisms controlling these levels are more finely tuned. We also report significant increases in brain, lung, and kidney NAD⁺ in the Cd38^−/− mouse, and provide the first experimental demonstration of the proximate relationship between CD38 and NAD⁺.

Section snippets

Materials and methods

Animals and tissue preparation. Male Cd38^−/− and male C57BL6/J adult mice were obtained from the Trudeau Institute (Saranac Lake, NY). Cd38^−/− mice were generated by gene targeting [20] and were backcrossed for 12 generations to C57BL/6J [18]. This study was approved by the Animal Care Committee at the University of Guelph and adhered to the standards set forth by the Canadian Council on Animal Care. For preparation of brain tissue, animals were anesthetized with Isofluorane (Fisher Scientific)

Results

The results of this study are illustrated in Fig. 1, Fig. 2. Fig. 1 shows the NAD⁺ concentrations in the measured organs. Heart NAD⁺(Fig. 1a) did not show a significant change due to genotype, while lung (Fig. 1b) (p < 0.001), kidney NAD⁺ (Fig. 1c) (p = 0.02), and brain NAD⁺ (Fig. 1d) concentrations were significantly (p < 0.001) increased in Cd38^−/− as compared to wild type mice. Fig. 2 shows the cADPR concentrations in the measured organs. Heart (Fig. 2a) and kidney (Fig. 2c) cADPR did not show a

Discussion

This is the first study to show significant decreases in levels of brain and lung cADPR in the Cd38^−/− mouse. Partida-Sanchez et al. (2001) previously measured levels of cADPR in these tissues and found them to be non-significantly decreased. While our levels of cADPR (with the exception of lung, discussed later) are comparable to the original report, the degree of variability in each group is reduced, which we believe is due to modifications that we have made to the fluorimetric cycling assay

Acknowledgment

This work was supported by funding from the Natural Sciences and Engineering Research Council (NSERC) of Canada.

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To the best of our knowledge, this is the first process in which a physiological decrease in CD38 is documented. CD38 is the major enzyme involved in NAD+ consumption: its levels markedly affect NAD+ content, as demonstrated by modulating its expression in cell lines [31], or in different organs (i.e. lung, kidney and brain) from Cd38−/− mice [32]. Here we observed that the genetic ablation of Cd38 per se strongly increases NAD+ levels also in BAT and WAT from mice kept at 30–22 °C: specifically, the absence of CD38 induced a more than doubled NAD+ content in BAT, whereas in WAT the effect was even more pronounced, with NAD+ being 6-fold higher in Cd38−/− mice (Fig. 2A, C).
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^☆: Abbreviations: cADPR, cyclic ADP-ribose; NAD⁺, nicotinamide adenine dinucleotide; PCA, perchloric acid; KOH, potassium hydroxide.

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Decreased cADPR and increased NAD+ in the Cd38−/− mouse☆

Abstract

Section snippets

Materials and methods

Results

Discussion

Acknowledgment

Leuk. Res.

Biochem. Biophys. Res. Commun.

Blood

J. Urol.

Biochem. Biophys. Res. Commun.

Brain Res.

Brain Res.

Blood

Anal. Biochem.

J. Biol. Chem.

Topology of CD38

Chem. Immunol.

Enzymatic functions and structures of CD38 and homologs

Chem. Immunol.

Coordinated regulation in human T cells of nucleotide-hydrolyzing ecto-enzymatic activities, including CD38 and PC-1. Possible role in the recycling of nicotinamide adenine dinucleotide metabolites

J. Immunol.

CD38 in T- and B-cell functions

Chem. Immunol.

NAD degradation and regulation of CD38 expression by human monocytes/macrophages

Eur. J. Biochem.

Decreased cADPR and increased NAD⁺ in the Cd38^−/− mouse☆