Cerebroventricular microinjection (CVMI) into adult zebrafish brain is an efficient misexpression method for forebrain ventricular cells.
Cerebroventricular microinjection (CVMI) into adult zebrafish brain is an efficient misexpression method for forebrain ventricular cells.
复制标题
DOI：
10.1371/journal.pone.0027395
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Brand M
中科院分区：
文献类型：
--
作者：
Kizil C;Brand M
斑马鱼（Danio rerio）具有在其生命的成年阶段在其大脑中产生新生神经元的显着能力，这一过程称为成年神经发生。这种能力依赖于心室祖细胞的增殖，与成年神经发生能力相当有限的哺乳动物大脑形成鲜明对比。因此，研究斑马鱼的大脑不仅有助于阐明脊椎动物物种中广泛的成年神经发生的分子机制，而且还有助于利用我们从这种硬骨鱼中学到的知识设计人类的治疗方法。然而，了解成年斑马鱼大脑中不同生物过程的细胞行为和分子程序需要允许操纵基因功能的技术。作为目前使用的斑马鱼中的错误表达技术的补充方法，如转基因方法或基于电穿孔的DNA递送，我们设计了一种脑室显微注射（CVMI）辅助敲除方案，该方案依赖于体内吗啉代寡核苷酸，其不需要电穿孔进行细胞摄取。这种快速的方法允许均匀和有效的基因敲除在斑马鱼脑的心室细胞，其中包含神经源性祖细胞。我们还提供了关于使用CVMI向大脑施用生长因子的数据-在我们的情况下是FGF 8，其调节心室细胞的增殖速率。在本文中，我们描述了CVMI方法，并讨论其在斑马鱼的潜在用途。
The teleost fish Danio rerio (zebrafish) has a remarkable ability to generate newborn neurons in its brain at adult stages of its lifespan-a process called adult neurogenesis. This ability relies on proliferating ventricular progenitors and is in striking contrast to mammalian brains that have rather restricted capacity for adult neurogenesis. Therefore, investigating the zebrafish brain can help not only to elucidate the molecular mechanisms of widespread adult neurogenesis in a vertebrate species, but also to design therapies in humans with what we learn from this teleost. Yet, understanding the cellular behavior and molecular programs underlying different biological processes in the adult zebrafish brain requires techniques that allow manipulation of gene function. As a complementary method to the currently used misexpression techniques in zebrafish, such as transgenic approaches or electroporation-based delivery of DNA, we devised a cerebroventricular microinjection (CVMI)-assisted knockdown protocol that relies on vivo morpholino oligonucleotides, which do not require electroporation for cellular uptake. This rapid method allows uniform and efficient knockdown of genes in the ventricular cells of the zebrafish brain, which contain the neurogenic progenitors. We also provide data on the use of CVMI for growth factor administration to the brain – in our case FGF8, which modulates the proliferation rate of the ventricular cells. In this paper, we describe the CVMI method and discuss its potential uses in zebrafish.
登录
查看更多内容
影响因子：
2.7
作者：
Davison, Jon M.;Akitake, Courtney M.;Parsons, Michael J.
通讯作者：
Parsons, Michael J.
影响因子：
2.8
作者：
Golberg, Alex;Rubinsky, Boris
通讯作者：
Rubinsky, Boris
影响因子：
2.3
作者：
Kim, Seongcheol;Radhakrishnan, Uvaraj P.;Jagadeeswaran, Pudur
通讯作者：
Jagadeeswaran, Pudur
影响因子：
2.7
作者：
Kizil, Caghan;Otto, Georg W.;Antos, Christopher L.
通讯作者：
Antos, Christopher L.
影响因子：
4.8
作者：
Asakawa, Kazuhide;Kawakami, Koichi
通讯作者：
Kawakami, Koichi