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| 斑马鱼早期胚胎血细胞分布特征 |
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秦思师1,2,3, 何秋霞4, 刘艳松1,2, 金瑶波1,2,3, 王丽娟1,2, 尤锋1,2, 谭训刚1,2
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1.中国科学院海洋研究所, 水产品种创制与高效养殖重点实验室, 中国科学院和山东省实验海洋生物学重点实验室, 海洋大科学中心, 山东 青岛, 266071;2.青岛海洋科技中心海洋生物学与生物技术功能实验室, 山东 青岛, 266273;3.中国科学院大学, 北京, 10049;4.齐鲁工业大学(山东省科学院)高新技术产业(中试)基地(山东省科学院留学人员创业园), 山东 济南, 250353
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| 摘要: |
| 斑马鱼是研究动物发育生物学的模式生物, 被用于研究血管血液疾病的机制及相关病理特征。为了解斑马鱼早期胚胎血液循环情况, 本文收集斑马鱼受精卵并培育至孵化, 在不同时间节点利用定量PCR分析血细胞、血管及血红蛋白相关基因的表达情况；同时, 使用邻联茴香胺对血红蛋白进行染色, 并对染色胚胎进行组织切片, 利用解剖镜和显微镜进行观察和拍照；将基因表达与血红蛋白染色分布相对应分析斑马鱼早期胚胎不同时期血液系统发育情况。定量PCR结果显示受精后12 h, 原始造血开始, 受精后24 h, 血管开始形成, 72 h之前斑马鱼血液循环系统已基本发育完全；而通过染色观察有一定的滞后, 在斑马鱼胚胎受精后30 h才可以观察到血红蛋白在卵黄囊中出现, 受精后48 h可在心脏观察到红细胞, 此时通过切片可以在躯干和体节中发现红细胞；通过切片还可以在72 h胚胎的脑部发现红细胞；受精后96 h斑马鱼胚胎血液循环基本发育成熟。本研究不仅仅从基因水平及蛋白水平对应描述了血管及血细胞(红细胞为主)的发育过程及分布特征, 也为通过简单检测方法分析血液相关疾病的病理及毒理特征奠定了基础。 |
| 关键词: 斑马鱼 胚胎 血液 |
| DOI：10.11759/hykx20240408001 |
| 分类号:Q78;Q17 |
| 基金项目:山东省自然科学基金(ZR2022MC129);齐鲁工业大学(山东省科学院)科教产先导专项(2020KJC-ZD10);国家自然科学基金(31672636);山东省自然科学基金联合基金(ZR2023LZL005) |
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| Distribution characteristics of blood cells in early zebrafish embryos |
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QIN Sishi1,2,3, HE Qiuxia4, LIU Yansong1,2, JIN Yaobo1,2,3, WANG Lijuan1,2, YOU Feng1,2, TAN Xungang1,2
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1.Key Laboratory of Aquatic Product Breeding and Efficient Breeding, Key Laboratory of Experimental Marine Biology of Chinese Academy of Sciences and Shandong Province, Ocean Science Center, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China;2.Qingdao Marine Science and Technology Center Laboratory of Marine Biology and Biotechnology Function, Qingdao, 266273, China;3.University of Chinese Academy of Sciences, Beijing, 10049, China;4.Qilu University of Technology (Shandong Academy of Sciences) High-tech Industry (Pilot) Base (Shandong Academy of Sciences Overseas Students Pioneer Park), Jinan, 250353, China
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| Abstract: |
| The zebrafish (Danio rerio) is a model organism for investigating animal developmental biology and is used to evaluate the mechanism and related pathological characteristics of vascular and blood diseases. Fertilized eggs of zebrafish were collected and incubated until hatching to determine blood circulation in early embryos. Quantitative polymerase chain reaction (PCR) was used to analyze the expression of genes related to blood cells, blood vessels, and hemoglobin at various developmental stages. Simultaneously, o-dianisidine was used to stain embryonic hemoglobin. Following staining, an anatomical lens and microscope were used to section, examine, and photograph the embryos. Gene expression data were compared with hemoglobin distribution to analyze the development of the blood system at different stages in early zebrafish embryos. Quantitative PCR results revealed that primitive hematopoiesis began at 12 h post-fertilization, blood vessel formation began at 24 h post-fertilization, and the blood circulation system was nearly fully developed by 72 h post-fertilization. However, staining observations showed a delay; red blood cells were first detected in the yolk sac of zebrafish embryos at 30 h post-fertilization, circulating blood in the heart at 48 h post-fertilization, and red blood cells in the trunk and somites by sectioning. Red blood cells can also be found in the brain 72 h post-fertilization of embryos by sectioning. Blood circulation in zebrafish embryos matured at 96 h post-fertilization. This study details the development timeline and distribution characteristics of blood vessels and blood cells (red blood cells mainly) at gene and protein levels, providing a foundation for analyzing pathological and toxicological features of blood-related diseases using simple detection methods. |
| Key words: zebrafish embryos blood |
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