期刊
JOURNAL OF NEUROSCIENCE METHODS
卷 283, 期 -, 页码 23-32出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jneumeth.2017.03.011
关键词
CLARITY; PARVALBUMIN; Schizophrenia; 3D analysis
Background: Novel tissue clearing technologies have, for the first time, made it possible to study intact tissue samples. This approach provides a tool for further clarifying findings from animal models of schizophrenia by studying parvalbumin-positive (PV+) interneuron density from a 3D perspective. New method: This study has developed an optimised CLARITY protocol, including an improved electrophoretic tissue clearing (ETC) chamber, an evaluation of antibody diffusion into cleared tissue slices, and a computational method for detecting PV+ interneurons in 3D. Results: A reduced PV+ interneuron density was found in both prelimbic and motor cortex regions of the Df(h15q13)/+ mice, while no changes were observed in the Df(h22q11)/+ mice. Comparison with existing method: The developed ETC chamber enables tissue clearing of variable tissue sizes while minimizing the resistance. It was found that a high concentration of primary and secondary antibodies were necessary for sufficient antibody staining of PV+ interneurons. Additionally, the developed computational method showed improved detection rates of interneurons compared to non -processed image stacks. Conclusion: Our optimization of the CLARITY technology and automated 3D counting of cells were found to be useful for quantification of PV+ interneuron density. The results may provide insight into understanding the pathophysiology underlying the phenotype observed in Df(h15q13)/+ mice. (C) 2017 Elsevier B.V. All rights reserved.
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