| Midkine-a通过限制心肌细胞总数的方式阻碍斑马鱼胚胎心脏生长 |
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| 引用本文： | 陈静,LuoJing,HitchcockPeter. Midkine-a通过限制心肌细胞总数的方式阻碍斑马鱼胚胎心脏生长[J]. 实验动物与比较医学, 2015, 23(5): 458-465 |
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| 作者姓名： | 陈静 LuoJing HitchcockPeter |
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| 作者单位： | 福建师范大学,美国密歇根大学安娜堡分校,美国密歇根大学安娜堡分校 |
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| 基金项目： | 国家自然科学基金项目（面上项目，重点项目，重大项目）;国家教育部回国人员科研启动基金 |
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| 摘 要： | 目的 探讨一种可溶性的外分泌因子Midkine-a在斑马鱼胚胎心脏发育过程中的功能。方法 在整体胚胎上做Midkine-a RNA的原位杂交实验；利用原有的转基因斑马鱼系Tg (pMidkine-a:EGFP)，动态观察胚胎从出生到心脏发育成形这一段时间心脏荧光表达情况；将原有的转基因斑马鱼体系 Tg (phsp:Midkine-a:EGFP) 胚胎进行热休克而过表达Midkine-a，观察胚胎心脏表型；利用Tg (pcmlc2:dsRed)鱼系胚胎的心肌细胞核带有红色荧光，能进行单个心脏心肌细胞计数这一特点，将杂合的Tg (phsp:Midkine-a:EGFP)鱼系与纯合的Tg (pcmlc2:dsRed)鱼系交配，以得到Tg (phsp:Midkine-a:EGFP/pcmlc2:dsRed)的杂合胚胎，对其进行热休克而过表达Midkine-a，计算每个胚胎心脏内心肌细胞的总数; 用吗啉寡聚核苷酸 （Morphonino，MO）阻碍新生胚胎内的Midkine-a mRNA 表达，观察胚胎心脏表型。结果 原位杂交试验证实Midkine-a在胚胎48hpf （hours post fertilization, 受精后小时，用来标记斑马鱼胚胎年龄）大时表达于心脏；转基因Tg (pMidkine-a:EGFP)胚胎在72hpf时其EGFP表达于心脏；Tg(phsp:Midkine-a:EGFP) 胚胎在过表达Midkine-a后心脏变小；吗啉寡聚核苷酸敲除Midkine-a对胚胎心脏发育无影响；最后在Tg (phsp:Midkine-a:EGFP/pcmlc2:dsRed)鱼系胚胎内过表达Midkine-a导致其单个心脏内心肌细胞数目变少，与其小心脏外形吻合。结论 Midkine-a在斑马鱼胚胎发育过程中表达于胚胎心脏；过表达Midkine-a导致胚胎心脏内心肌细胞总数减少及心脏变小；敲除Midkine-a则对胚胎心脏发育无影响。
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| 关 键 词： | 斑马鱼胚胎；心脏发育；Midkine-a；心肌细胞池 |
| 收稿时间： | 2015-04-27 |
| 修稿时间： | 2015-07-16 |
Midkine-a inhibits zebrafish embryonic heart growth by limiting the cardiomyocyte pool |
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| CHEN Jing,LUO Jing and Hitchcock Peter. Midkine-a inhibits zebrafish embryonic heart growth by limiting the cardiomyocyte pool[J]. Laboratory Animal and Comparative Medicine, 2015, 23(5): 458-465 |
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| Authors: | CHEN Jing LUO Jing Hitchcock Peter |
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| Affiliation: | Fujian Normal University, Fuzhou 350117, China;Kellogg Eye Center of University of Michigan-Ann Arbor, Ann Arbor, US 48103;Kellogg Eye Center of University of Michigan-Ann Arbor, Ann Arbor, US 48103 |
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| Abstract: | Objective To investigate the functional role of a soluble secretion factor Midkine-a in the process of zebrafish embryonic heart development. Methods Whole-mount in situ hybridization was perform to detect whether Midkine-a is expressed in the embryonic heart; in the transgenic embryonic heart of Tg(pMidkine-a:EGFP), the expression of EGFP in the heart was monitored ; Multiple heat shock regimens were applied to Tg(phsp:Midkine-a:EGFP) embryos in order to overexpress Midkine-a, and the phenotype of the heart was observed; The heterozygous Tg(phsp:Midkine-a:EGFP) fish were crossed with homozygous Tg(pcmlc2:dsRed) fish, which specifically expresses RFP in the nucleoli of embryonic cardiomyocyte to facilitate myocyte number counting, to get Tg(phsp:Midkine-a:EGFP/pcmlc2:dsRed) embryos, and the total number of cardiomyocytes in their heart were counted and compared to the controls when the heat shock induced overexpression of Midkine-a; Morphonino runing down of Midkine-a was performed to watch the heart phenotype. Results Midkine-a is expressed in the zebrafish embryonic heart during development; overexpression of Midkine-a leads to smaller heart and reduced total number of cardiomyoctes in a single heart, which might associates with the smaller heart phenotype. Morphonilo running down of Midkine-a has no effects upon heart development. Conclusions Midkine-a impedes zebrafish embryonic heart growth by limiting its cardiomyocyte pool. |
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| Keywords: | Zebrafish embryo Heart development midkine-a Cardiomyocyte pool |
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