The Shisa family is a type of single-transmembrane adaptor proteins containing an N-terminal cysteine-rich domain and a proline-rich C-terminal region. Nine Shisa subfamily genes have been proposed in most vertebrates; however, some of them might be species-specific. The number of Shisa genes present in zebrafish remains unclear. The purpose of this study was to investigate the evolutionary relationships among Shisa family genes in zebrafish (TU strain) using phylogenetic and syntenic analyses. The function of shisa-2 was preliminarily examined via CRISPR/Cas13d-mediated knockdown. Following identification in zebrafish,10 Shisa family genes, namely shisa-1, 2, 3, 4, 5, 6, 7, 8, 9a, and 9b, were classified into three main clades and six subclades. Their encoding proteins contained a cysteine-rich N-terminal domain and a proline-rich C-terminal region containing different motifs. A specific syntenic block containing atp8a2 and shisa-2 was observed to be conserved across all species. Furthermore, all these genes were found to be expressed during embryogenesis. shisa-2 was expressed in the presomitic mesoderm, somites, and so on. shisa-2 was identified as a regulator of the expression of the somite formation marker mesp-ab. Overall, our study provides new insights into the evolution of Shisa family genes and the control of shisa-2 over the convergent extension cells of somatic precursors in zebrafish.
This study evaluated the effect of Lentinus edodes fermentation (LEF) on digestive enzyme activity, antioxidant capacity and morphology of the liver and intestine in largemouth bass (Micropterus salmoides) fed high plant protein diets (HPPD). LEF was supplemented in HPPD with 0 g kg−1 (LEF0), 10 g kg−1 (LEF1), 20 g kg−1 (LEF2), 30 g kg−1 (LEF3), 40 g kg−1 (LEF4), 50 g kg−1 (LEF5), respectively, and then the six diets were fed to largemouth bass with a body weight of 28.8 ± 0.05 g for eight weeks. Juvenile fish were randomized into 6 groups and each group had 4 replicates with 40 fish. Dietary LEF supplementation alleviated the liver inflammatory reaction of largemouth bass caused by HPPD and improved liver morphology. Goblet cells multiplied and the gut muscle layer thickened after LEF supplementation. The LEF significantly increased amylase activity in the liver and intestine of largemouth bass in individual experimental groups. The LEF could increase the activity of catalase in the liver and intestine of largemouth bass (p < 0.05). The content of malondialdehyde was significantly lower than that in the control group (p < 0.05). Dietary LEF supplementation had no significant effect on the intestinal flora of largemouth bass. These findings imply that LEF supplementation can reduce liver inflammation, enhance intestinal tissue morphology, and eventually benefit largemouth bass health.
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