Stereotaxic Surgery for Excitotoxic Lesion of Specific Brain Areas in the Adult Rat
Stereotaxic Surgery for Excitotoxic Lesion of Specific Brain Areas in the Adult Rat
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DOI：
10.3791/4079
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发表时间：
2012-07-01
影响因子：
1.2
通讯作者：
Kaufer, Daniela
中科院分区：
文献类型：
--
作者：
Kirby, Elizabeth D.;Jensen, Kelly;Kaufer, Daniela
哺乳动物的许多行为功能，包括啮齿动物和人类，主要是由分散的脑区调节的。辨别不同大脑区域的行为功能或其他实验结果的常用方法是实施局部功能消融。在人类中，经常对局部脑损伤的患者群体进行缺陷研究，希望揭示受损区域的潜在功能。在啮齿类动物中，人们可以通过实验诱导特定大脑区域的损伤。病变可以通过几种方式完成。电解损伤可引起局部损伤，但也会损伤多种类型的细胞以及来自恰好在损伤部位附近的其他大脑区域的穿越纤维。使用细胞类型特异性启动子的诱导遗传技术也可以实现位点特异性靶向。这些技术很复杂，并不总是根据目标大脑区域的不同而实用。相比之下，使用立体定向手术的兴奋性毒性病变是损伤兴奋性神经元的最可靠和实用的方法之一，而不会损伤局部胶质细胞或穿过纤维。在这里，我们提出了一种将兴奋毒素n -甲基- d -天冬氨酸（NMDA）立体定向注入基底外侧杏仁核复合体的方案。根据解剖学指示，我们应用立体坐标来确定目标大脑区域的位置，并在目标上方放置一根注射针。然后我们将兴奋毒素注入大脑，导致附近神经元的兴奋性毒性死亡。虽然我们选择的实验对象是一只老鼠，但同样的方法可以应用于其他哺乳动物，只要对设备和坐标进行适当的调整。该方法可用于多种脑区，包括基底外侧杏仁核（1-6）、其他杏仁核（6,7）、海马(8)、内嗅皮质(9)和前额皮质（10）。它也可用于注入诸如病毒载体的生物化合物（1,11）。基本的立体定向技术也可以用于植入更永久的渗透泵，允许更长时间地暴露于感兴趣的化合物。
Many behavioral functions in mammals, including rodents and humans, are mediated principally by discrete brain regions. A common method for discerning the function of various brain regions for behavior or other experimental outcomes is to implement a localized ablation of function. In humans, patient populations with localized brain lesions are often studied for deficits, in hopes of revealing the underlying function of the damaged area. In rodents, one can experimentally induce lesions of specific brain regions.Lesion can be accomplished in several ways. Electrolytic lesions can cause localized damage but will damage a variety of cell types as well as traversing fibers from other brain regions that happen to be near the lesion site. Inducible genetic techniques using cell-type specific promoters may also enable site-specific targeting. These techniques are complex and not always practical depending on the target brain area. Excitotoxic lesion using stereotaxic surgery, by contrast, is one of the most reliable and practical methods of lesioning excitatory neurons without damaging local glial cells or traversing fibers.Here, we present a protocol for stereotaxic infusion of the excitotoxin, N-methyl-D-aspartate (NMDA), into the basolateral amygdala complex. Using anatomical indications, we apply stereotaxic coordinates to determine the location of our target brain region and lower an injection needle in place just above the target. We then infuse our excitotoxin into the brain, resulting in excitotoxic death of nearby neurons. While our experimental subject of choice is a rat, the same methods can be applied to other mammals, with the appropriate adjustments in equipment and coordinates.This method can be used on a variety of brain regions, including the basolateral amygdala(1-6), other amygdala nuclei(6, 7), hippocampus(8), entorhinal cortex(9) and prefrontal cortex(10). It can also be used to infuse biological compounds such as viral vectors(1, 11). The basic stereotaxic technique could also be adapted for implantation of more permanent osmotic pumps, allowing more prolonged exposure to a compound of interest.