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ADP-dependent glucokinase (ADPGK) plays an important role instead of hexokinase in regulating energy metabolism via the Embden–Meyerhof–Parnas Pathway. And energy provided via glycolysis promotes testis development and spermatogenesis. In this study, 466 Hu sheep were screened for mutations in the ADPGK gene to examine the association of the ADPGK gene polymorphisms with the testis traits and spermatogenesis. The NC_056060.1: g.31295 C > T SNP was found in the 3′-UTR region, resulting in two genotypes CC and TC type with genotypic frequencies of 0.66 and 0.34, respectively. This mutation was significantly associated with testis weight, testis long circumference, testis short girth, epididymis weight, and sperm concentration (p < 0.05). Moreover, TC genotype individuals had an increased tendency in the expression of the ADPGK gene and had significant reproductive performance advantages compared with CC genotype individuals in the study. And compared with the small testes (<50 g), the ADPGK gene expression of big testes (>160 g) increased significantly. This indicates an association between the ADPGK gene and reproductive organ parameters and sperm count in selected Hu sheep breed, and this SNP may serve as an effective DNA molecular marker for marker-assisted selection in Hu sheep breeding programs. [ABSTRACT FROM AUTHOR]

The fatty acid content and the localization and expression of phospholipase A 2 (PLA 2) in the testis of Hu sheep were investigated. A total of 18 six-month-old Hu sheep were divided into small group (S, with left testis weight < 50 g), medium group (M, with left testis weight among 90–110 g), and large group (L, with left testis weight >160 g), which had six individuals each. The expression of PLA 2 in testicular tissues of different sizes was analyzed by immunohistochemistry, RT-qPCR, and Western blot. The fatty acid profile was detected by gas chromatography. Immunohistochemical labeling determined that PLA 2 protein was expressed in the Leydig and Sertoli cells of testis, and the immunohistochemical average optional density in the S group was significantly greater than the L group (P < 0.05). RT-qPCR and Western blot analysis showed that PLA 2 in the S group was greater than that in the L group (P < 0.05). Docosahexaenoic acid, ω-3 polyunsaturated fatty acid (PUFA), and total PUFA content in the testis of the L group were significantly less than those of the S and M groups (P < 0.01). This study showed that PLA 2 content in the S group was greater than that in the L group. • Phospholipase A2 was widely distributed in the testis of Hu sheep. • The expression of phospholipase A2 in the small testis group was increased. • The ω-3 polyunsaturated fatty acids were decreased in large testis. [ABSTRACT FROM AUTHOR]

Energy supply is crucial for testicular development. Nevertheless, the specific alterations in the energy metabolic pathways that affect testicular development have not been extensively investigated. This study aimed to investigate the variations in metabolites and alterations in energy metabolic pathways in the testes of Hu sheep with different developmental status at 6 months of age. Twelve rams with similar body weights but distinct testis developmental status were selected among 345 Hu sheep based on testis size and histomorphology results, and they were divided into the well-developed (L group, n = 6) and developmentally delayed group (S group, n = 6). A total of 660 metabolites were identified via widely targeted metabolic analysis. Among 148 differentially expressed metabolites, 78 were up-regulated and 70 were downregulated in the L group compared with the S group. Functional enrichment analysis indicated that a significant proportion of the identified differential metabolites was implicated in energy metabolism-related pathways. Moreover, the L group exhibited significantly higher expression levels of genes involved in glycolysis (GLTU8 and LDH), TCA (PDHA2, CS and IDH3G), gluconeogenesis (PCK1), pentose phosphate (G6PD), and fatty acid degradation (GK, ACSL1, FABP3, CPT1 and CTP2). The activity enzymes such as citrate synthase, pyruvate dehydrogenase, and lactate dehydrogenase also increased in the L group. In summary, this observation implied that the augmentation of energy metabolic pathways plays a crucial role in facilitating testicular development. The upregulation of energy metabolic pathways collectively facilitates the testicular development in Hu sheep., Competing Interests: Declaration of competing interest The authors declare that there are no potential conflicts of interest related to this study., (Copyright © 2024. Published by Elsevier Inc.)