Liquid storage of spermatozoa is important for artificial insemination and herd genetic breeding. However, the extended time of storage inducing the rapid decline in spermatozoa quality limits the development of this technology. The molecular mechanisms underlying liquid storage of spermatozoa remain largely unexplored. In this study, the effects of liquid storage on functional quality of spermatozoa were assessed in goat (Capra hircus). The time-dependent decline in spermatozoa motility showed a strong correlation with the significant increase in apoptosis. Moreover, apoptosis-related ultrastructural changes were observed, especially the defects in mitochondria. A significant decrease in mitochondrial membrane potential and changes in the expression of mitochondrial apoptosis-related proteins indicated mitochondrial dysfunction and mitochondrial apoptotic pathway activation. Notably, the abnormally high level of reactive oxygen species (ROS) caused by liquid storage resulted in oxidative damage to mitochondria and accelerated mitochondria-dependent apoptosis, as demonstrated by the addition of ROS scavenger N-acetylcysteine. Furthermore, critical differentially expressed proteins involved in mitochondria-dependent apoptosis and antioxidant defense were identified and profiled by quantitative proteomic analysis, facilitating the understanding of molecular regulation of ROS-induced mitochondria-dependent apoptosis. These outcomes provide insights into the mechanisms underlying liquid storage of goat spermatozoa and enhance the progress of semen storage technology.

No abstract

No abstract

Water temperature has a major influence on the host innate immune defence and the infectivity of pathogens in ectothermic teleosts. Toll‐like receptors (TLRs) are the first and well‐characterized innate immune receptors that are conserved in vertebrates. However, little is known about the effect of temperature variation on TLRs in fish species. In this study, we used adult zebrafish as a research model to investigate the effect of water temperature on TLRs. Whole genome searches identified 20 TLR homologue genes in zebrafish. Multiple sequence alignment and protein structure analysis revealed the conserved domains for these TLR proteins. To identify TLR genes related to temperature variation, TLR family genes from 12 species with different body temperatures were assigned to conduct phylogenetic analyses. Based on the phylogenetic relationships, TLR3, TLR4, TLR5 and TLR20~21 were selected as candidate genes. Immunostimulation data indicated that TLR3, TLR5, and TLR21 were more sensitive to temperature variation and their expression levels were affected in response to pathogen stimulation. Taken together, our results provide a new opportunity to understand the roles of temperature on host innate immune response in fishes and have broader implications for disease prevention in aquaculture.

No abstract