硬化蛋白基因在淇河鲫成鱼不同肌间骨相邻肌组织的表达差异分析

为探究硬化蛋白(sclerostin, SOST)基因在淇河鲫肌间骨不同分布位置中的作用，以淇河鲫成鱼为研究对象，对其肌间骨的形态与分布进行了统计，并在此基础上利用qRT-PCR技术检测SOST基因在淇河鲫背部和尾部肌肉中的mRNA表达，通过Western印迹和免疫组织化学技术检测SOST蛋白的表达定位情况。结果显示：淇河鲫肌间骨包括髓弓小骨和脉弓小骨，髓弓小骨为56～58根，位于淇河鲫背部肌肉的肌隔中，脉弓小骨27～28根，分布于泄殖孔之后的肌隔中。背部肌肉SOST的mRNA表达水平显著高于尾部肌肉（P < 0.05）。与mRNA表达水平相比，背部肌肉和尾部肌肉中SOST蛋白呈现相同表达趋势。免疫组织化学结果显示SOST在肌隔组织中特异性表达，并且在背部肌肉中的表达强烈，尾部肌肉中没有阳性反应。上述结果表明，SOST在淇河鲫肌间骨的背部和尾部肌肉中存在差异性表达，从而影响肌隔组织中肌间骨的骨化，对背部和尾部肌间骨形态发生产生影响。

Differentially Expression of SOST Gene in the Muscle Tissues Located between Different Intermuscular Bones in Qihe Crucian Carp Carassius auratus

To study the roles of SOST gene in different distribution position of intermuscular bones in Qihe crucian carp (Carassius auratus), the number and morphology of intramuscular bones were counted and viewed. Based on the morphological analysis of intermuscular bones, quantitative real-time PCR (qRT-PCR), Western blotting and immunohistochemistry methods were used to examine the mRNA and protein expression of SOST in dorsal and caudal muscles with intermuscular bones. The results showed that there were only epineural and epipleural bones in Qihe Crucian carp, and the number of epineural bones which located in the myoseptum of dorsal muscles were between 56 and 58, and epipleural bones which located in the myoseptum after the cloacal orifice ranged from 27 to 28. The expression level of SOST in dorsal muscles was significantly higher than that of caudal muscles. The protein expression of SOST in dorsal and caudal muscles exhibited the similar trend to that of the mRNA. Immunohistochemistry analysis proved that there were significantly positive reaction in the myoseptum of dorsal muscles, and no positive signals were detected in the myoseptum of caudal muscles. The above results suggested that SOST was expressed significantly different in dorsal and caudal muscle associated with intramuscular bones, which might influence the ossification of intermusuclar bones and lead to the morphological variation of intermusular bones in Qihe crucian carp.