Generation, Maintenance, and Identification of Germ-Free Zebrafish Models from Larvae to Juvenile Stages

作者全名：Jia, Pan -Pan; Liu, Xiang-Ru; Wu, Ming-Fei; Li, Yan; Zhang, Lan -Chen; Pei, De-Sheng

作者地址：[Jia, Pan -Pan; Liu, Xiang-Ru; Wu, Ming-Fei; Li, Yan; Zhang, Lan -Chen; Pei, De-Sheng] Chongqing Med Univ, Sch Publ Hlth, Chongqing, Peoples R China

通信作者：Pei, DS (通讯作者)，Chongqing Med Univ, Sch Publ Hlth, Chongqing, Peoples R China.

来源：JOVE-JOURNAL OF VISUALIZED EXPERIMENTS

ESI学科分类：Multidisciplinary

WOS号：WOS:001237200900001

JCR分区：Q3

影响因子：1.2

年份：2024

卷号：　

期号：206

开始页：10

结束页：21

文献类型：Article

关键词：　

摘要：Zebrafish serve as valuable models for research on growth, immunity, and gut microbiota due to their genomic similarities with mammals, transparent embryos developed in a relatively clean chorion environment, and extremely rapid development of larvae compared to rodent models. Germ-free (GF) zebrafish (Danio rerio) are crucial for evaluating pollutant toxicity and establishing human-like disease models related to microbial functions. In comparison to conventionally raised (CR) models (fish in common husbandry), GF zebrafish allow for more accurate manipulation of the host microbiota, aiding in determining the causal relationship between microorganisms and hosts. Consequently, they play a critical role in advancing our understanding of these relationships. However, GF zebrafish models are typically generated and researched during the early life stages (from embryos to larvae) due to limitations in immune function and nutrient absorption. This study optimizes the generation, maintenance, and identification of early GF zebrafish models without feeding and with long-term feeding using GF food (such as Artemia sp., brine shrimp). Throughout the process, daily sampling and culture were performed and identified through multiple detections, including plates and 16S rRNA sequencing. The aseptic rate, survival, and developmental indexes of GF zebrafish were recorded to ensure the quality and quantity of the generated models. Importantly, this study provides details on bacterial isolation and infection techniques for GF fish, enabling the efficient creation of GF fish models from larvae to juvenile stages with GF food support. By applying these procedures in biomedical research, scientists can better understand the relationships between intestinal bacterial functions and host health.

基金机构：Chongqing Medical University Talent Project [R4014, R4020]; National Natural Science Foundation of China (NSFC) [32200386]; Chongqing Postdoctoral Innovation Mentor Studio [X7928]; Program of China -Sri Lanka Joint Center for Water Technology Research and Demonstration by Chinese Academy of Sciences (CAS) /China -Sri Lanka Joint Center for Education and Research by CAS

基金资助正文：We sincerely thank the support from Chongqing Medical University Talent Project (No. R4014 to DSP and R4020 to PPJ) , National Natural Science Foundation of China (NSFC, No.32200386 to PPJ) , Chongqing Postdoctoral Innovation Mentor Studio (X7928 DSP) , and Program of China -Sri Lanka Joint Center for Water Technology Research and Demonstration by Chinese Academy of Sciences (CAS) /China -Sri Lanka Joint Center for Education and Research by CAS.