cd99l2基因调控斑马鱼白细胞组织间的迁移机制

doi:10.16288/j.yczz.22-193

遗传 ›› 2022, Vol. 44 ›› Issue (9): 798-809.doi: 10.16288/j.yczz.22-193

cd99l2基因调控斑马鱼白细胞组织间的迁移机制

卢荆澳^1,²(), 黄春燕^1,², 林芷茵^1,², 唐政^1,², 马宁^1,²(), 黄志斌³()

1. 南方医科大学基础医学院组织胚胎学教研室,广州 510515
2. 国家药监局化妆品安全评价重点实验室,南方医科大学,广州 510515
3. 华南理工大学医学院发育生物学与再生医学团队,广州 510006

收稿日期:2022-06-13 修回日期:2022-08-17 出版日期:2022-09-20 发布日期:2022-08-19
通讯作者: 马宁,黄志斌 E-mail:18609668060@163.com;mn1228@smu.edu.cn;huangzhb1986@scut.edu.cn
作者简介:卢荆澳,本科,专业方向：人体解剖与组织胚胎学。E-mail: 18609668060@163.com
基金资助:
国家重点研发计划编号(2018YFA0801000);国家自然科学基金青年基金项目资助(31401231)

The role of the cd99l2 gene on leukocyte interstitial migration in zebrafish

Jing’ao Lu^1,²(), Chunyan Huang^1,², Zhiyin Lin^1,², Zheng Tang^1,², Ning Ma^1,²(), Zhibin Huang³()

1. Department of Histology and Embryology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China
2. NMPA Key Laboratory for Safety Evaluation of Cosmetics, Southern Medical University, Guangzhou 510515, China
3. Division of Development Biology & Regenerative Medicine, South China University of Technology, Guangzhou 510006, China;

Received:2022-06-13 Revised:2022-08-17 Online:2022-09-20 Published:2022-08-19
Contact: Ma Ning,Huang Zhibin E-mail:18609668060@163.com;mn1228@smu.edu.cn;huangzhb1986@scut.edu.cn
Supported by:
Supported by the National Key Research and Development Program of China(2018YFA0801000);the National Natural Science Foundation of China No(31401231)

摘要/Abstract

摘要：

白细胞在个体发育、组织再生、先天及适应性免疫过程中都发挥了重要作用,而白细胞迁移至感染或创伤区域是其发挥免疫应答功能的必要条件。CD99L2作为黏附分子在白细胞外渗过程中发挥了重要作用,但其是否参与白细胞组织间的迁移尚不明确。本研究通过TALEN技术敲除斑马鱼(Danio rerio) cd99l2基因,发现cd99l2基因缺失后并不影响斑马鱼正常发育。尾鳍损伤后,cd99l2基因突变体在创伤组织处募集的中性粒细胞与巨噬细胞数目显著减少,进一步研究发现mfap4在cd99l2基因突变体中表达显著降低,这可能是影响巨噬细胞向损伤组织处迁移的原因之一。通过标记血管内皮细胞与中性粒细胞及巨噬细胞的转基因斑马鱼品系,发现受精后60 h野生型或突变体斑马鱼中性粒细胞和巨噬细胞通过组织间迁移至创伤组织处,表明cd99l2基因参与了白细胞在组织间的迁移。利用RNA-seq分析发现,cd99l2基因突变体中RNA转录、蛋白质折叠及p450通路等被富集。黏附分子调控转录过程及信号传递在以往研究中已被多次证实。据此,本研究推测Cd99l2作为黏附分子参与了白细胞迁移过程中的级联信号通路。综上所述,本研究通过对斑马鱼的研究,发现了cd99l2基因在白细胞迁移过程中新的作用,有望对炎症免疫异常相关疾病提供理论基础。

关键词: 斑马鱼cd99l2基因, 白细胞迁移, mfap4, 信号级联通路

Abstract:

Leukocytes play an essential role in ontogeny, tissue regeneration, and innate and adaptive immunity. The migration of leukocytes to the infected or traumatized areas is necessary for their immune response function. As an adhesion molecule, CD99L2 is crucial in the extravasation of leukocytes, however, its role in the interstitial migration of leukocytes remains unclear. In this study, the cd99l2 gene was knock-out by TALEN (transcription activator-like effector nuclease) in zebrafish and discovered that the deletion had no effect on zebrafish development. The number of granulocytes and macrophages recruited to the wounded tissue was significantly reduced in the cd99l2 mutants following caudal fin damage. Further research revealed that the expression of mfap4 was drastically decreased in the cd99l2 mutants, which may be one of the reasons that affect the migration of macrophages to the wound site. Moreover,transgenic lines with labeled vasculature, neutrophils and macrophages demonstrated that neutrophils and macrophages migrate throughout the interstitial space to the wound tissue in both wild-type and mutant zebrafish at 60 hours post-fertilization, indicating that the cd99l2 gene is involved in the interstitial migration of leukocytes. Finally, RNA transcription, protein folding, and the P450 pathway were enriched in cd99l2 mutants by RNA-seq analysis. Previous research had demonstrated that the regulation of transcription and signal transduction could be affected by adhesion molecules, which may suggest that the cd99l2 gene is involved in the cascade signaling pathway of leukocyte migration as an adhesion molecule. In conclusion, this study uncovered a novel function of the cd99l2 gene in the process of leukocyte migration in zebrafish, which is expected to provide a theoretical foundation for inflammatory and immune-related diseases.

Key words: zebrafish cd99l2 gene, leukocyte migration, mfap4 gene, cascade signaling pathway

卢荆澳, 黄春燕, 林芷茵, 唐政, 马宁, 黄志斌. cd99l2基因调控斑马鱼白细胞组织间的迁移机制[J]. 遗传, 2022, 44(9): 798-809.

Jing’ao Lu, Chunyan Huang, Zhiyin Lin, Zheng Tang, Ning Ma, Zhibin Huang. The role of the cd99l2 gene on leukocyte interstitial migration in zebrafish[J]. Hereditas(Beijing), 2022, 44(9): 798-809.

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基因名称	引物序列(5′→3′)
ef1a	F: TACTTCTCAGGCTGACTGTG
ef1a	R: ATCTTCTTGATGTATGCGCT-3
cd99l2	F: CTACGACACCATGGCTGAGACT
cd99l2	R: CTTGTGGCTCCTGTGCGACTAC
lyz	F: AAAGCAGGTTTAAGACCCAC
lyz	R: CCAGGTTTCCCATGATTTCAG
mpx	F: AGAGACTGATAGAGATTCCATCC
mpx	R: CCAGGTTTCCCATGATTTCAG
mfap4	F: GTTTACACCATCTATCCAGCC
mfap4	R: GTTCTCTAGTCCCAGCCA

cd99l2基因调控斑马鱼白细胞组织间的迁移机制

The role of the cd99l2 gene on leukocyte interstitial migration in zebrafish

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