Single-Cell RNA-Seq and Histological Analysis Reveals Dynamic Lrig1 Expression During Salivary Gland Development.

IF 4.5 2区 生物学 Q2 CELL BIOLOGY
Shumin Liu, Yuanyuan Li, Delan Huang, Ming Liu, Xinye Zhang, Hui Zhao, Huan Liu, Qiuhui Li, Zhi Chen
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引用次数: 0

Abstract

The development of the salivary gland (SG) is a complex process regulated by multiple signaling pathways in a spatiotemporal manner. Various stem/progenitor cell populations and respective cell lineages are involved in SG morphogenesis and postnatal maturation. Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) has been identified as critical regulator of stem cells by virtue of its ability to restrain stem cell proliferation, indicating its potential role in the development of several maxillofacial tissues and in the regulation of the quiescence in adult tissues. This study aimed to investigate the expression pattern and functions of Lrig1 in the developing and mature murine submandibular gland (SMG). To accomplish this objective, we collected the murine SMGs at different developmental stages and examined the expression pattern and levels of Lrig1 with qRT-PCR, immunofluorescent (IF) and RNAscope staining. We observed that Lrig1 was widely distributed in both epithelial and mesenchymal cells throughout embryonic and neonatal stages, with specific localization in the more mature epithelium. Furthermore, through single-cell RNA sequencing (scRNA-Seq) and IF techniques, we confirmed that LRIG1 is highly concentrated along with SMG progenitor markers in acinar and basal cells. Additionally, transcription factors (TFs) that could regulate LRIG1 expression were predicted from JASPAR databases and their motifs were identified by the UCSC browser's BLAT tool. Gene Ontology (GO) enrichment analyses on postnatal day 5 (PN5) scRNA-Seq data also provided insights into Lrig1's functions in SG development. Finally, we also conducted in vitro experiments on a human salivary gland (HSG) cell line to assess LRIG1's impact on HSG proliferation and migration, as well as its potential upstream regulatory TFs. Taken together, our study reveals that LRIG1 plays a vital role in SG development.

单细胞 RNA-Seq 和组织学分析揭示唾液腺发育过程中 Lrig1 的动态表达
唾液腺(SG)的发育是一个复杂的过程,受多种信号通路的时空调控。各种干/祖细胞群和各自的细胞系都参与了唾液腺的形态发生和出生后的成熟。富亮氨酸重复序列和免疫球蛋白样结构域1(LRIG1)因其抑制干细胞增殖的能力而被认为是干细胞的关键调节因子,这表明它在多种颌面部组织的发育和成人组织的静止调节中具有潜在作用。本研究旨在探讨 Lrig1 在发育和成熟的小鼠颌下腺(SMG)中的表达模式和功能。为了实现这一目标,我们采集了处于不同发育阶段的小鼠SMG,并通过qRT-PCR、免疫荧光(IF)和RNA镜染色检测了Lrig1的表达模式和水平。我们观察到,Lrig1广泛分布于胚胎和新生儿期的上皮细胞和间质细胞中,并在较成熟的上皮细胞中有特异性定位。此外,通过单细胞 RNA 测序(scRNA-Seq)和 IF 技术,我们证实 LRIG1 与 SMG 祖先标记物一起高度集中在尖状细胞和基底细胞中。此外,我们还从 JASPAR 数据库中预测了可调控 LRIG1 表达的转录因子(TFs),并通过 UCSC 浏览器的 BLAT 工具确定了它们的基序。对出生后第 5 天(PN5)scRNA-Seq 数据进行的基因本体(GO)富集分析也有助于深入了解 Lrig1 在 SG 发育中的功能。最后,我们还对人类唾液腺(HSG)细胞系进行了体外实验,以评估 LRIG1 对 HSG 增殖和迁移的影响及其潜在的上游调控 TFs。综上所述,我们的研究揭示了 LRIG1 在唾液腺发育过程中的重要作用。
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来源期刊
CiteScore
14.70
自引率
0.00%
发文量
256
审稿时长
1 months
期刊介绍: The Journal of Cellular Physiology publishes reports of high biological significance in areas of eukaryotic cell biology and physiology, focusing on those articles that adopt a molecular mechanistic approach to investigate cell structure and function. There is appreciation for the application of cellular, biochemical, molecular and in vivo genetic approaches, as well as the power of genomics, proteomics, bioinformatics and systems biology. In particular, the Journal encourages submission of high-interest papers investigating the genetic and epigenetic regulation of proliferation and phenotype as well as cell fate and lineage commitment by growth factors, cytokines and their cognate receptors and signal transduction pathways that influence the expression, integration and activities of these physiological mediators. Similarly, the Journal encourages submission of manuscripts exploring the regulation of growth and differentiation by cell adhesion molecules in addition to the interplay between these processes and those induced by growth factors and cytokines. Studies on the genes and processes that regulate cell cycle progression and phase transition in eukaryotic cells, and the mechanisms that determine whether cells enter quiescence, proliferate or undergo apoptosis are also welcomed. Submission of papers that address contributions of the extracellular matrix to cellular phenotypes and physiological control as well as regulatory mechanisms governing fertilization, embryogenesis, gametogenesis, cell fate, lineage commitment, differentiation, development and dynamic parameters of cell motility are encouraged. Finally, the investigation of stem cells and changes that differentiate cancer cells from normal cells including studies on the properties and functions of oncogenes and tumor suppressor genes will remain as one of the major interests of the Journal.
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