Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation
Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation
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DOI：
10.3791/50412
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发表时间：
2013-07-01
影响因子：
1.2
通讯作者：
Alvarez-Bolado, Gonzalo
中科院分区：
文献类型：
--
作者：
Haddad-Tvolli, Roberta;Szabo, Nora-Emke;Alvarez-Bolado, Gonzalo
对发育中的哺乳动物大脑的特定区域进行遗传修饰是一种非常有效的实验方法。然而，产生新的小鼠突变体往往是令人沮丧的缓慢。已经表明，在子宫内发育的小鼠大脑的手术后存活率是合理的。尽管如此，这种手术的结果几乎只针对发育中大脑的最表层和最容易接近的部分，即皮层。丘脑是一个更窄、更中间的区域，已经证明更难瞄准。转染到更深的核团，特别是下丘脑的核团，可能是最具挑战性的，因此报道的结果很少。在这里，我们展示了一种通过电穿孔靶向整个下丘脑神经上皮或其部分（下丘脑区域）进行转染的方法。我们的方法的关键是更长的麻醉时间，在第三脑室注射，以及适当的电极类型和位置。此外，我们显示了最凹陷的下丘脑核，乳头体的靶向和随后的组织学分析的结果。
Genetic modification of specific regions of the developing mammalian brain is a very powerful experimental approach. However, generating novel mouse mutants is often frustratingly slow. It has been shown that access to the mouse brain developing in utero with reasonable post-operatory survival is possible. Still, results with this procedure have been reported almost exclusively for the most superficial and easily accessible part of the developing brain, i.e. the cortex. The thalamus, a narrower and more medial region, has proven more difficult to target. Transfection into deeper nuclei, especially those of the hypothalamus, is perhaps the most challenging and therefore very few results have been reported. Here we demonstrate a procedure to target the entire hypothalamic neuroepithelium or part of it ( hypothalamic regions) for transfection through electroporation. The keys to our approach are longer narcosis times, injection in the third ventricle, and appropriate kind and positioning of the electrodes. Additionally, we show results of targeting and subsequent histological analysis of the most recessed hypothalamic nucleus, the mammillary body.