XLαs通过影响线粒体功能调控颅骨发育的研究

doi:10.13701/j.cnki.kqyxyj.2024.07.004

口腔医学研究 ›› 2024, Vol. 40 ›› Issue (7): 587-592.DOI: 10.13701/j.cnki.kqyxyj.2024.07.004

XLαs通过影响线粒体功能调控颅骨发育的研究

左祎怡, 何青^*

口颌系统重建与再生全国重点实验室,口腔生物医学教育部重点实验室,口腔医学湖北省重点实验室,武汉大学口腔医(学)院湖北武汉 430079

收稿日期:2024-04-02 出版日期:2024-07-28 发布日期:2024-07-24
通讯作者: *何青,E-mail: qing.he@whu.edu.cn
作者简介:左祎怡(1998~ ),女,江西上饶人,硕士在读,研究方向:骨发育。
基金资助:
国家自然科学基金(编号:82072483)

Regulatory Effects of XLαs on Cranial Bone Development through Influencing Mitochondrial Function

ZUO Yiyi, HE Qing^*

State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan 430079, China.

Received:2024-04-02 Online:2024-07-28 Published:2024-07-24

摘要/Abstract

摘要： 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。

关键词: 超大型刺激性G蛋白α亚单位, 颅骨发育, 矿化, 线粒体

Abstract: 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 expression 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 unmineralized 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 novel therapeutic strategy for treating cranial developmental anomalies associated with GNAS mutations.

Key words: extra-large stimulatory G protein alpha subunit, cranial development, mineralization, mitochondria

左祎怡, 何青. XLαs通过影响线粒体功能调控颅骨发育的研究[J]. 口腔医学研究, 2024, 40(7): 587-592.

ZUO Yiyi, HE Qing. Regulatory Effects of XLαs on Cranial Bone Development through Influencing Mitochondrial Function[J]. Journal of Oral Science Research, 2024, 40(7): 587-592.

参考文献

[1] Turan S, Bastepe M. The GNAS complex locus and human diseases associated with loss-of-function mutations or epimutations within this imprinted gene [J]. Horm Res Paediatr, 2013, 80(4):229-241.
[2] Yang W, Zuo Y, Zhang N, et al. GNAS locus: bone related diseases and mouse models [J]. Front Endocrinol (Lausanne), 2023, 14: 1255864.
[3] Xie Y, Chen X, Xie Y, et al. GNAS gene mutations affecting XLΑS and bone health: A long neglected relationship [J]. Clin Genet, 2023, 104(3):279-286.
[4] Maduro AI, Pinto Saraiva A, Pimenta Rodrigues O, et al. Albright’s hereditary osteodystrophy: an entity to recognize [J]. Rheumatology (Oxford), 2022, 61(11): e356-e357.
[5] Linglart A, Levine MA, Jüppner H. Pseudohypoparathyroidism [J]. Endocrinol Metab Clin North Am, 2018, 47(4): 865-888.
[6] Leclercq V, Benoit V, Lederer D, et al. Case report: An infantile lethal form of Albright hereditary osteodystrophy due to a GNAS mutation [J]. Clin Case Rep, 2018, 6(10): 1933-1940.
[7] Lei R, Zhang K, Wei Y, et al. G-protein α-subunit Gsα is required for craniofacial morphogenesis [J]. PLoS One, 2016, 11(2): e0147535.
[8] Xu R, Liu Y, Zhou Y, et al. GNAS loss causes chondrocyte fate conversion in cranial suture formation [J]. J Dent Res, 2022, 101(8): 931-941.
[9] Plagge A, Gordon E, Dean W, et al. The imprinted signaling protein XLαs is required for postnatal adaptation to feeding [J]. Nat Genet, 2004, 36(8): 818-826.
[10] Mead TJ. Alizarin red and Alcian blue preparations to visualize the skeleton[M]. Apte SS. ADAMTS proteases: Methods and protocols. New York, NY: Springer, 2020. 207-212.
[11] Liu B, Lu Y, Wang Y, et al. A protocol for isolation and identification and comparative characterization of primary osteoblasts from mouse and rat calvaria [J]. Cell Tissue Bank, 2019, 20(2): 173-182.
[12] Wang Y, Tian H, Chen X. The distinct role of the extra-large G protein ɑ-subunit XLɑs [J]. Calcif Tissue Int, 2020, 107(3): 212-219.
[13] He Q, Bouley R, Liu Z, et al. Large G protein α-subunit XLαs limits clathrin-mediated endocytosis and regulates tissue iron levels in vivo [J]. Proc Natl Acad Sci U S A, 2017, 114(45): E9559-E9568.
[14] Galea GL, Zein MR, Allen S, et al. Making and shaping endochondral and intramembranous bones[J]. Dev Dyn, 2021, 250(3):414-449.
[15] Liao J, Huang Y, Wang Q, et al. Gene regulatory network from cranial neural crest cells to osteoblast differentiation and calvarial bone development [J]. Cell Mol Life Sci, 2022, 79(3): 158.
[16] Salhotra A, Shah HN, Levi B, et al. Mechanisms of bone development and repair [J]. Nat Rev Mol Cell Biol, 2020, 21(11): 696-711.
[17] Zheng CX, Sui BD, Qiu XY, et al. Mitochondrial regulation of stem cells in bone homeostasis [J]. Trends Mol Med, 2020, 26(1): 89-104.
[18] Shen L, Hu G, Karner CM. Bioenergetic metabolism in osteoblast differentiation [J]. Curr Osteoporos Rep, 2022, 20(1): 53-64.
[19] Dobson PF, Dennis EP, Hipps D, et al. Mitochondrial dysfunction impairs osteogenesis, increases osteoclast activity, and accelerates age related bone loss [J]. Sci Rep, 2020, 10(1): 11643.

XLαs通过影响线粒体功能调控颅骨发育的研究

Regulatory Effects of XLαs on Cranial Bone Development through Influencing Mitochondrial Function

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