Modified Golgi-Cox method for micrometer scale sectioning of the whole mouse brain

One of the major challenges of connectomics is obtaining a physical map of the neurons that comprise a circuit and the sites within the whole mouse brain. However, there is no report that addresses the preparation of whole mouse brain tissue for microsectioning. In this paper, such tissue is prepared by a modified Golgi-Cox method in which the staining time is prolonged to half a year, the darkening solution is changed to 1% LiOH, and the brain is embedded in resin. Projections of several coronal sections are reconstructed by serial 1-μm sectioning and simultaneous imaging of the specimen. This approach ensures that the stained neurons are present throughout the whole mouse brain from superficial to deep layers and that the neuronal soma and traces of the processes can be distinguished in local magnification.

[1]  G. Moralí,et al.  Long-term evaluation of cytoarchitectonic characteristics of prefrontal cortex pyramidal neurons, following global cerebral ischemia and neuroprotective melatonin treatment, in rats , 2008, Neuroscience Letters.

[2]  E. Ramon‐Moliner,et al.  A tungstate modification of the Golgi-Cox method. , 1958, Stain technology.

[3]  B. Hyman,et al.  Neonatal neuronal overexpression of glycogen synthase kinase-3β reduces brain size in transgenic mice , 2002, Neuroscience.

[4]  A. B. Silva-Gómez,et al.  Decreased dendritic spine density on prefrontal cortical and hippocampal pyramidal neurons in postweaning social isolation rats , 2003, Brain Research.

[5]  R. Sidman,et al.  Atlas of the Mouse Brain and Spinal Cord , 1971 .

[6]  J. D. Macklis,et al.  Multipotent neural precursors can differentiate toward replacement of neurons undergoing targeted apoptotic degeneration in adult mouse neocortex. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Facundo Valverde,et al.  Golgi Atlas of the Postnatal Mouse Brain , 2004 .

[8]  J. Hutsler,et al.  Does microwaving enhance the Golgi methods? A quantitative analysis of disparate staining patterns in the cerebral cortex , 2003, Journal of Neuroscience Methods.

[9]  John H. Martin Neuroanatomy: Text and Atlas , 1989 .

[10]  E. Roubos,et al.  Ultrastructural and immunocytochemical characterization of the rat non-preganglionic Edinger-Westphal nucleus. , 2009, General and comparative endocrinology.

[11]  W. Denk,et al.  Serial Block-Face Scanning Electron Microscopy to Reconstruct Three-Dimensional Tissue Nanostructure , 2004, PLoS biology.

[12]  Bryan Kolb,et al.  A method for vibratome sectioning of Golgi–Cox stained whole rat brain , 1998, Journal of Neuroscience Methods.

[13]  B. Decourt,et al.  Expression analysis of neuroleukin, calmodulin, cortactin, and Rho7/Rnd2 in the intact and injured mouse brain. , 2005, Brain research. Developmental brain research.

[14]  George Paxinos,et al.  The Mouse Brain in Stereotaxic Coordinates , 2001 .

[15]  Håkan Johansson,et al.  Modern Techniques in Neuroscience Research , 1999, Springer Berlin Heidelberg.

[16]  Edmund M. Glaser,et al.  Analysis of thick brain sections by obverse—Reverse computer microscopy: Application of a new, high clarity Golgi—Nissl stain , 1981, Journal of Neuroscience Methods.

[17]  Y. Goshima,et al.  Increased proximal bifurcation of CA1 pyramidal apical dendrites in sema3A mutant mice , 2009, The Journal of comparative neurology.

[18]  G. Rosoklija,et al.  Optimization of Golgi methods for impregnation of brain tissue from humans and monkeys , 2003, Journal of Neuroscience Methods.

[19]  Arthur W. Toga,et al.  The informatics of a C57BL/6J mouse brain atlas , 2007, Neuroinformatics.