Zhao Ran, Zhu Jianqiang, Li Shaowu, Sun Zhipeng, Zhang Tingting, Wang Jing, Zheng Xianhu, Kuang Youyi, Wang Di
Key Laboratory of Aquatic Animal Diseases and Immune Technology of Heilongjiang Province, Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070, China.
Antioxidants (Basel). 2025 Apr 8;14(4):443. doi: 10.3390/antiox14040443.
The presence of intermuscular bones severely affects the edibility and value-added processing of crucian carp (), becoming a constraint to the high-quality development of its industry. Our previous study identified as the key osteogenic regulator and successfully developed a new crucian carp strain lacking intermuscular bones (WUCI) using CRISPR/Cas9 technology. To accelerate its industrialization, we comprehensively assessed WUCI's growth performance, hematological parameters, antioxidant capacity, innate immunity, and disease resistance. The results demonstrated that the WUCI exhibited significant growth performance compared to the wild-type crucian carp (WT), with significantly higher weight gain (WG) and specific growth rate (SGR) ( < 0.05) from one month to four months of age. The α-amylase (α-AL) activity of the liver and intestines of WUCI was significantly higher than that of WT. WUCI also displayed enhanced intestinal antioxidant capacity, with superoxide dismutase (SOD) and catalase (CAT) activities significantly higher than those in WT ( < 0.05). The malondialdehyde (MDA) content in the spleen of WUCI was significantly lower than that of WT ( < 0.05); no differences were observed in the liver and intestines ( > 0.05). Additionally, hepatic acid phosphatase (ACP) activity in WUCI was significantly higher than that in WT ( < 0.05). In contrast, splenic ACP and intestinal alkaline phosphatase (ALP) activities were significantly lower than those in WT ( < 0.05). Notably, the iron concentration in the serum was significantly higher in WUCI than in the WT ( < 0.05). Meanwhile, WUCI exhibited significantly lower a expression of , , and mRNA in the liver compared to WT ( < 0.05), while mRNA expression was significantly higher ( < 0.05). Routine blood tests revealed significantly lower WBC in WUCI compared to that of WT ( < 0.05). Following an challenge, WT demonstrated a rapid transcriptional induction of pro-inflammatory cytokines (, , ) and immunoregulatory mediators (, ), with mRNA levels reaching maximal expression at 24 h post-infection (hpi) followed by progressive attenuation. In contrast, WUCI exhibited a delayed immune activation profile characterized by the peak expression of , , , and transcripts after 72 hpi, with the maximum transcript abundance remaining lower than corresponding peak values observed in WT at 24 hpi. Finally, we observed that the mortality rate of WUCI was slightly higher post infection when compared to WT, but was not significant ( > 0.05). In conclusion, this study provides a comprehensive evaluation of WUCI, revealing its distinct growth advantages, physiological adaptations, and immune function, presenting its potential for aquaculture breeding applications.
肌间骨的存在严重影响鲫鱼的可食用性和增值加工,成为其产业高质量发展的制约因素。我们之前的研究确定[具体基因名称未给出]为关键的成骨调节因子,并利用CRISPR/Cas9技术成功培育出一种新的无肌间骨鲫鱼品系(WUCI)。为加速其产业化,我们全面评估了WUCI的生长性能、血液学参数、抗氧化能力、先天免疫和抗病性。结果表明,与野生型鲫鱼(WT)相比,WUCI表现出显著的生长性能,在1月龄至4月龄时,其增重率(WG)和特定生长率(SGR)显著更高(P<0.05)。WUCI肝脏和肠道的α-淀粉酶(α-AL)活性显著高于WT。WUCI还表现出增强的肠道抗氧化能力,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性显著高于WT(P<0.05)。WUCI脾脏中的丙二醛(MDA)含量显著低于WT(P<0.05);肝脏和肠道中未观察到差异(P>0.05)。此外,WUCI肝脏中的酸性磷酸酶(ACP)活性显著高于WT(P<0.05)。相比之下,脾脏ACP和肠道碱性磷酸酶(ALP)活性显著低于WT(P<0.05)。值得注意的是,WUCI血清中的铁浓度显著高于WT(P<0.05)。同时,与WT相比,WUCI肝脏中[具体基因名称未给出]、[具体基因名称未给出]和[具体基因名称未给出]mRNA的表达显著降低(P<0.05),而[具体基因名称未给出]mRNA表达显著升高(P<0.05)。常规血液检测显示,WUCI的白细胞数量显著低于WT(P<0.05)。在进行[具体病原体未给出]攻击后,WT表现出促炎细胞因子([具体细胞因子名称未给出]、[具体细胞因子名称未给出]、[具体细胞因子名称未给出])和免疫调节介质([具体介质名称未给出]、[具体介质名称未给出])的快速转录诱导,mRNA水平在感染后24小时(hpi)达到最大表达,随后逐渐衰减。相比之下,WUCI表现出延迟的免疫激活特征,其[具体基因名称未给出]、[具体基因名称未给出]、[具体基因名称未给出]和[具体基因名称未给出]转录本在72 hpi达到峰值表达,最大转录本丰度仍低于WT在24 hpi时观察到的相应峰值。最后,我们观察到,感染[具体病原体未给出]后,WUCI的死亡率略高于WT,但不显著(P>0.05)。总之,本研究对WUCI进行了全面评估,揭示了其独特的生长优势、生理适应性和免疫功能,展示了其在水产养殖育种应用中的潜力。