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结合腔室组件的3D打印和多尺度光学成像对小鼠脑丘-丘脑-皮质脑片进行改造

Modification of a Colliculo-thalamocortical Mouse Brain Slice, Incorporating 3-D printing of Chamber Components and Multi-scale Optical Imaging.

作者信息

Slater Bernard J, Fan Anthony Y, Stebbings Kevin A, Saif M Taher A, Llano Daniel A

机构信息

Neuroscience Program, University of Illinois Urbana-Champaign.

Mechanical Science and Engineering, University of Illinois Urbana-Champaign.

出版信息

J Vis Exp. 2015 Sep 18(103):53067. doi: 10.3791/53067.

Abstract

The ability of the brain to process sensory information relies on both ascending and descending sets of projections. Until recently, the only way to study these two systems and how they interact has been with the use of in vivo preparations. Major advances have been made with acute brain slices containing the thalamocortical and cortico-thalamic pathways in the somatosensory, visual, and auditory systems. With key refinements to our recent modification of the auditory thalamocortical slice(1), we are able to more reliably capture the projections between most of the major auditory midbrain and forebrain structures: the inferior colliculus (IC), medial geniculate body (MGB), thalamic reticular nucleus (TRN), and the auditory cortex (AC). With portions of all these connections retained, we are able to answer detailed questions that complement the questions that can be answered with in vivo preparations. The use of flavoprotein autofluorescence imaging enables us to rapidly assess connectivity in any given slice and guide the ensuing experiment. Using this slice in conjunction with recording and imaging techniques, we are now better equipped to understand how information processing occurs at each point in the auditory forebrain as information ascends to the cortex, and the impact of descending cortical modulation. 3-D printing to build slice chamber components permits double-sided perfusion and broad access to networks within the slice and maintains the widespread connections key to fully utilizing this preparation.

摘要

大脑处理感觉信息的能力依赖于上行和下行投射系统。直到最近,研究这两个系统及其相互作用的唯一方法一直是使用活体标本。在包含躯体感觉、视觉和听觉系统中丘脑皮质和皮质丘脑通路的急性脑切片方面已经取得了重大进展。通过对我们最近对听觉丘脑皮质切片的改进(1)进行关键优化,我们能够更可靠地捕捉大多数主要听觉中脑和前脑结构之间的投射:下丘(IC)、内侧膝状体(MGB)、丘脑网状核(TRN)和听觉皮层(AC)。由于保留了所有这些连接的部分,我们能够回答一些详细问题,这些问题补充了可以用活体标本回答的问题。黄素蛋白自发荧光成像的使用使我们能够快速评估任何给定切片中的连接性,并指导后续实验。将该切片与记录和成像技术结合使用,我们现在更有能力了解随着信息向上传递到皮层时,听觉前脑的每个点是如何进行信息处理的,以及下行皮质调制的影响。使用3D打印来制造切片室组件允许双面灌注,并能广泛接触切片内的网络,同时保持充分利用该标本的关键广泛连接。

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本文引用的文献

1
An auditory colliculothalamocortical brain slice preparation in mouse.在小鼠中制备听觉丘脑海马皮质脑片。
J Neurophysiol. 2014 Jan;111(1):197-207. doi: 10.1152/jn.00605.2013. Epub 2013 Oct 9.
3
Topography and physiology of ascending streams in the auditory tectothalamic pathway.听觉顶盖丘脑通路上升束的地形和生理学。
Proc Natl Acad Sci U S A. 2010 Jan 5;107(1):372-7. doi: 10.1073/pnas.0907873107. Epub 2009 Dec 15.
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Attentional modulation of thalamic reticular neurons.丘脑网状核神经元的注意力调节
J Neurosci. 2006 Apr 19;26(16):4444-50. doi: 10.1523/JNEUROSCI.5602-05.2006.
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A visual thalamocortical slice.一个视觉丘脑皮质切片。
Nat Methods. 2006 Feb;3(2):129-34. doi: 10.1038/nmeth849.
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Auditory thalamocortical synaptic transmission in vitro.体外听觉丘脑皮质突触传递
J Neurophysiol. 2002 Jan;87(1):361-84. doi: 10.1152/jn.00549.2001.

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