在肌肉平滑肌细胞中的表面局部和总催产素受体的量化
Yingye Fang1, Erin L Reinl2, Audrey Liu2
1Department of Bioengineering, University of Washington, Seattle, WA, 98109, USA.
Heliyon
|February 22, 2024
概括
催产素受体 (OXTR) 的遗传变异在人类肌肉细胞中显示出细胞表面局部变化. 这证实了先前的发现,并强调了定量流细胞计对于在本地细胞环境中研究OXTR的有用性.
科学领域:
- 生殖生物学 生殖生物学
- 分子内分泌学分子内分泌学
- 细胞生理学 细胞生理学
背景情况:
- 催产素受体 (OXTR) 对于子宫收缩性至关重要.
- 在OXTR的遗传变异可以影响其功能.
- 以前的研究表明,在HEK293T细胞中,一些OXTR变异的细胞表面局部发生变化.
研究的目的:
- 量化OXTR表达在细胞表面和总细胞在人类肌肉细胞的平滑肌肉细胞 (HMSMCs).
- 调查OXTR变异在更原生HMSMC环境中的局部化.
- 为了验证使用CRISPR编辑的HA标记OXTR进行定量测量.
主要方法:
- 利用CRISPR编辑引入HA标签进入HMSMCs内源性OXTR基因.
- 采用定量流细胞计量来测量总和细胞表面的OXTR水平.
- 在HMSMC中对外表达的野生类型和变异OXTRs与HA和GFP标签进行比较分析.
主要成果:
- 在HMSMC中,大约有55,000个OXTR和4,900个细胞表面OXTR在每个细胞中表达.
- 野生类型和变种OXTRs的总蛋白质表达是可比的.
- 两种先前确定的OXTR变异在HMSMC中表现出细胞表面局部减少,反映了HEK293T细胞的发现.
结论:
- 在原生人类肌肉细胞模型中证实了OXTR变异的差异细胞表面定位.
- 定量流细胞计是测量HMSMC中OXTR表达的合适技术.
- 这些发现对了解OXTR基因变异对子宫收缩能力的功能影响有意义.
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