目的:本研究旨在评估为分离α,beta,和成年小鼠胰岛的δ细胞,将其应用扩展到新生和老年小鼠的胰岛。此外,它试图检查小鼠胰腺内分泌胰岛细胞在整个出生后发育过程中的转录组动力学,并验证这些细胞群体中与年龄相关的改变。
方法:我们利用β细胞上CD71和δ细胞上CD24的高表面表达来FACS纯化α,beta,和来自新生儿(1周龄)的三角洲细胞,成人(12周龄),和老(18个月大)小鼠。对这些纯化的细胞群体进行了大量RNA测序,随后的生物信息学分析包括差异基因表达,代表性过高,和交叉分析。
结果:Alpha,beta,使用与成年小鼠相同的方法成功地FACS纯化了新生和老年小鼠的δ细胞。我们对α的年龄相关转录变化的分析,beta,和δ细胞群显示,在从新生小鼠到成年小鼠的过渡过程中,细胞周期减少,神经元样特征过程增加。从成年小鼠发展到老年小鼠,我们确定了与衰老(炎症)相关的炎症基因特征,包括β-2微球蛋白和主要组织相容性复合体(MHC)I类表达的增加.
结论:我们的研究证明了我们的细胞分选技术在从不同年龄的小鼠胰岛中纯化内分泌亚群的有效性。我们为更好地了解内分泌胰腺衰老提供了宝贵的资源,并确定了具有β-2微球蛋白和MHCI类表达增加的炎症基因标签作为旧α的常见标志,beta,和三角洲细胞,与免疫反应调节和年龄相关的糖尿病的潜在影响。
OBJECTIVE: This study aimed to evaluate the efficacy of a purification method developed for isolating alpha, beta, and delta cells from pancreatic islets of adult mice, extending its application to islets from newborn and aged mice. Furthermore, it sought to examine transcriptome dynamics in mouse pancreatic endocrine islet cells throughout postnatal development and to validate age-related alterations within these cell populations.
METHODS: We leveraged the high surface expression of CD71 on beta cells and CD24 on delta cells to FACS-purify alpha, beta, and delta cells from newborn (1-week-old), adult (12-week-old), and old (18-month-old) mice. Bulk RNA sequencing was conducted on these purified cell populations, and subsequent bioinformatic analyses included differential gene expression, overrepresentation, and intersection analysis.
RESULTS: Alpha, beta, and delta cells from newborn and aged mice were successfully FACS-purified using the same method employed for adult mice. Our analysis of the age-related transcriptional changes in alpha, beta, and delta cell populations revealed a decrease in cell cycling and an increase in neuron-like features processes during the transition from newborn to adult mice. Progressing from adult to old mice, we identified an inflammatory gene signature related to aging (inflammaging) encompassing an increase in β-2 microglobulin and major histocompatibility complex (MHC) Class I expression.
CONCLUSIONS: Our study demonstrates the effectiveness of our cell sorting technique in purifying endocrine subsets from mouse islets at different ages. We provide a valuable resource for better understanding endocrine pancreas aging and identified an inflammaging gene signature with increased β-2 microglobulin and MHC Class I expression as a common hallmark of old alpha, beta, and delta cells, with potential implications for immune response regulation and age-related diabetes.