47 Effect of a long-term, high-fat diet on metabolic health and oocyte quality of an outbred (Swiss) versus inbred (C57BL/6N) mouse strain

Abstract

Maternal metabolic disorders like obesity and diabetes type II are known to affect reproductive physiology, ultimately leading to poor fertility. The oocyte and embryo are extremely vulnerable during the periconceptional period to metabolic stressors, leading to disappointing fertility results. Most mouse model research regarding obesity and Western type diets has been performed on the inbred C57BL/6 strain. However, inbred strains are often linked with decreased fertility. Relying only on inbred strains might also limit translation to human (outbred) physiology. To further explore this, we compared the inbred C57BL/6N to an outbred Swiss strain. Five-week-old Swiss (N = 30) and C57BL/6N (B6) (N = 29) mice were fed a control (CTRL) or a high-fat (HF) diet for 13 weeks. Diets differed in percentage of fat (10% v. 60%). Body weight gain, serum profile (nonesterified fatty acids, cholesterol, and triglycerides), and oocyte quality were studied. Mature oocytes were collected after hormonal stimulation (IP injection of 10 IU of pregnant mare serum gonadotropin followed by 10 IU of hCG 48 h later). To study oocyte quality, Bodipy (lipid droplets), JC-1 (mitochondrial membrane potential), and Cell-Rox Deep Red stainings were performed, as well as transmission electron microscopy to examine mitochondrial structures. All data were analysed using the t-test. In comparison with the CTRL group, the HF diet increased body weight by 18.09 and 27.87% in Swiss and B6, respectively. The HF significantly increased blood cholesterol levels (103.5 v. 143.1 mg/dL in Swiss mice, 141.8 v. 185.4 mg/dL in B6 mice) in both strains, and tended to increase blood nonesterified fatty acids (P = 0.053) and triglycerides (P = 0.075) only in Swiss but not in B6 mice. Oocytes collected from the HF diet group contained a larger total volume of lipid droplets (P < 0.05) in both strains compared with controls. The mitochondrial membrane potential and Cell-Rox Deep Red were significantly increased (P < 0.05) in oocytes of Swiss mice, but not B6 mice, fed a HF diet. Transmission electron microscopy images from HF oocytes showed mitochondria with abnormal morphology, low electron density, and rose petal appearance, resulting in significantly increased mitochondrial abnormalities in Swiss mice on the HF diet (P < 0.05). In B6 mice, both CTRL and HF oocytes contained high proportions of abnormal mitochondria compared with the CTRL group of the Swiss mice, explaining the lack of HF diet effects in B6 oocyte ultrastructure. We conclude that a HF diet has a significant effect on both metabolic health and oocyte quality. However, the Swiss model seems more sensitive to a Western type diet insult, making it more suitable for research focusing on metabolic health and oocyte quality than the B6 strain. The HF diet-exposed Swiss mice showed differences (compared with CTRL) in their serum profile. Alterations in mitochondrial activity, structures, and oxidative stress were induced by HF diet in the Swiss mice and not the B6, although B6 oocytes also showed higher lipid droplet accumulation. Furthermore, even the B6 mice that were fed a normal control diet showed deviant oocyte quality, clearly shown by morphological signs of lower quality and mitochondrial abnormalities.