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. 2021 Oct 29;67(5):319-326.
doi: 10.1262/jrd.2021-080. Epub 2021 Aug 19.

Supplementing media with NAD+ precursors enhances the in vitro maturation of porcine oocytes

Charley-Lea Pollard  1 , Zamira Gibb  2 , Azelle Hawdon  1   3 , Aleona Swegen  2   4 , Christopher G Grupen  1
Affiliations

Supplementing media with NAD+ precursors enhances the in vitro maturation of porcine oocytes

Charley-Lea Pollard et al. J Reprod Dev. .

Abstract

In vitro maturation (IVM) is an important reproductive technology used to produce embryos in vitro. However, the developmental potential of oocytes sourced for IVM is markedly lower than those matured in vivo. Previously, NAD+-elevating treatments have improved oocyte quality and embryo development in cattle and mice, suggesting that NAD+ is important during oocyte maturation. The aim of this study was to examine the effects of nicotinic acid (NA), nicotinamide (NAM) and nicotinamide mononucleotide (NMN) on oocyte maturation and subsequent embryo development. Porcine oocytes from small antral follicles were matured for 44 h in a defined maturation medium supplemented with NA, NAM and resveratrol or NMN. Mature oocytes were artificially activated and presumptive zygotes cultured for 7 days. Additionally, oocytes were matured without treatment then cultured for 7 days with NMN. Supplementing the IVM medium with NA improved maturation and blastocyst formation while NAM supplementation improved cleavage rates compared with untreated controls. Supplementing the IVM or embryo culture media with NMN had no effect on maturation or embryo development. The results show that supplementing the maturation medium with NA and NAM improved maturation and developmental potential of porcine oocytes.

Keywords: Embryo development; In vitro maturation; Niacin; Oocyte quality; Pig.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Fig. 1.
Fig. 1.
NAD+ biosynthetic pathways. Two of the three pathways through which NAD+ can be synthesised in cells include the Preiss-Handler pathway and more commonly through the salvage of other metabolites. Nicotinic acid is consumed in feed and is the initial metabolite for the Preiss-Handler pathway. NAD+ is consumed by Sirtuins and Poly-ADP-ribose-polymerases, the first reaction in the salvage pathway. The metabolites in white boxes represent metabolites in the Preiss-Handler pathway while the metabolites in the light grey boxes represent those from the salvage pathway. NAD+ in the black box represents the end point for both NAD+ biosynthetic pathways.
Fig. 2.
Fig. 2.
The effect of NA at low doses on the rates of oocyte maturation, cleavage, blastocyst formation and blastocyst hatching. Bars labelled with different letters indicate statistical differences (P < 0.05).
Fig. 3.
Fig. 3.
The effect of NA at high doses on (A) the rates of oocyte maturation, cleavage, blastocyst formation and blastocyst hatching, and (B) total blastocyst cell number. Bars and plots labelled with different letters indicate statistical differences (P < 0.05).
Fig. 4.
Fig. 4.
The effect of NAM and Res, alone and in combination, on (A) the rates of oocyte maturation, cleavage, blastocyst formation and blastocyst hatching, and (B) total blastocyst cell number. Bars and plots labelled with different letters indicate statistical differences (P < 0.05).
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