目的:本研究旨在探究超大型刺激性G蛋白α亚单位(extra-large stimulatory G protein alpha subunit,XLαs)在小鼠颅骨发育和成骨分化中的功能及分子机制.方法:通过构建XLαs第一外显子敲除的小鼠模型,运用阿尔新蓝茜素红染色技术分析颅骨表型变化.从小鼠颅骨中分离原代颅骨成骨细胞,进行体外培养并诱导成骨分化,同时通过实时荧光定量聚合酶链反应和RNA测序技术对基因表达及其功能进行分析.结果:实验结果显示,XLαs敲除小鼠颅缝宽度增加(P＜0.05),颅骨未矿化区域明显增多(P＜0.001),且成骨分化能力受抑制(P＜0.01).RNA测序进一步揭示XLαs缺失影响了线粒体功能.结论:本研究证实了 XLαs在调节颅骨发育和成骨分化中的关键作用,特别是通过影响线粒体功能来发挥其调控效应.这一发现为针对GNAS突变导致的颅骨发育异常提供了新的治疗策略,具有潜在的临床应用价值.
Regulatory Effects of XLαs on Cranial Bone Development through Influencing Mitochondrial Function
Objective:To investigate the role and underlying molecular mechanism of extra-large stimulatory G protein alpha subunit(XLαs)in cranial bone development.Methods:Utilizing a genetically engineered mouse model lacking the specific exon 1 of XLαs,cranial morphology were analyzed through alician blue and alizarin red staining.Primary cranial osteoblasts were isolated and cultured in vitro,and induced for osteogenic differentiation.Gene ex-pression patterns and functional analysis were assessed through quantitative RT-PCR and RNA-seq.Results:Knockout of XLαs in mice led to notably increased cranial suture width(P＜0.05),expanded presence of unminer-alized areas within cranial bones(P＜0.001),and a significant reduction in osteogenic differentiation(P＜0.01).Further,RNA-seq analysis revealed that XLαs deficiency disrupted mitochondrial function.Conclusion:XLαs plays a critical regulatory role in the cranial bone development by modulating mitochondrial function,which provides a no-vel therapeutic strategy for treating cranial developmental anomalies associated with GNAS mutations.
extra-large stimulatory G protein alpha subunitcranial developmentmineralizationmitochondria
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