Department of Zoology and Physiology, University of Wyoming, Laramie, Wyoming.
Department of Zoology and Physiology, University of Wyoming, Laramie, Wyoming
J Neurophysiol. 2015 Jan 1;113(1):400-7. doi: 10.1152/jn.00672.2014. Epub 2014 Oct 15.
The Xenopus tadpole optic tectum is a multisensory processing center that receives direct visual input as well as nonvisual mechanosensory input. The tectal neurons that comprise the optic tectum are organized into layers. These neurons project their dendrites laterally into the neuropil where visual inputs target the distal region of the dendrite and nonvisual inputs target the proximal region of the same dendrite. The Xenopus tadpole tectum is a popular model to study the development of sensory circuits. However, whole cell patch-clamp electrophysiological studies of the tadpole tectum (using the whole brain or in vivo preparations) have focused solely on the deep-layer tectal neurons because only neurons of the deep layer are visible and accessible for whole cell electrophysiological recordings. As a result, whereas the development and plasticity of these deep-layer neurons has been well-studied, essentially nothing has been reported about the electrophysiology of neurons residing beyond this layer. Hence, there exists a large gap in our understanding about the functional development of the amphibian tectum as a whole. To remedy this, we developed a novel isolated brain preparation that allows visualizing and recording from all layers of the tectum. We refer to this preparation as the "horizontal brain slice preparation." Here, we describe the preparation method and illustrate how it can be used to characterize the electrophysiology of neurons across all of the layers of the tectum as well as the spatial pattern of synaptic input from the different sensory modalities.
非洲爪蟾幼体的视顶盖是一个多感官处理中心,它接收直接的视觉输入和非视觉机械感觉输入。构成视顶盖的顶盖神经元组织成层。这些神经元将其树突横向投射到神经胶质中,其中视觉输入靶向树突的远端区域,而非视觉输入靶向同一树突的近端区域。非洲爪蟾幼体视顶盖是研究感觉回路发育的常用模型。然而,使用全脑或在体制备的幼体视顶盖全细胞膜片钳电生理研究仅集中于深层视顶盖神经元,因为只有深层的神经元可见且可用于全细胞膜片钳电生理记录。结果,尽管这些深层神经元的发育和可塑性已经得到了很好的研究,但关于位于该层之外的神经元的电生理学基本上没有任何报道。因此,我们对整个两栖动物视顶盖的功能发育的理解存在很大的差距。为了解决这个问题,我们开发了一种新的分离脑制备方法,可以对视顶盖的所有层进行可视化和记录。我们将这种制备方法称为“水平脑切片制备”。在这里,我们描述了该制备方法,并说明了如何使用它来表征视顶盖所有层的神经元的电生理学,以及来自不同感觉模态的突触输入的空间模式。