Jan Baumgart1, Nadine Grebe2. 1. Institute for Microscopic Anatomy and Neurobiology, University Medical Center of the Johannes Gutenberg-University Mainz, 55131 Mainz, Germany; Central Laboratory Animal Facility, University Medical Center of the Johannes Gutenberg-University Mainz, 55131 Mainz, Germany. Electronic address: jan.baumgart@unimedizin-mainz.de. 2. Central Laboratory Animal Facility, University Medical Center of the Johannes Gutenberg-University Mainz, 55131 Mainz, Germany. Electronic address: greben@uni-mainz.de.
Abstract
BACKGROUND: In utero electroporation is a fast an efficient tool to specifically address gene expression in the murine central nervous system. This technique was originally established in ICR/CD-1 outbred mice. Neuroanatomical differences between the different mouse strains and variations in gestation length require the optimization of the conditions for each strain to avoid severe complications. Furthermore the relevant position information is currently only scarcely standardized and not always easy to transfer to C57BL/6 mice. NEW METHOD: In this study we present an improved method for in utero electroporation of C57BL/6 including a detailed atlas that allows for specific and efficient in vivo transfection. Further we introduce histogram analysis as a tool for neural migration assays. RESULTS: We report individually adapted conditions for in utero electroporation in C57BL/6 mice that differ from the previously published data for ICR/CD-1 mice. Furthermore, this article outlines a detailed angle-map that allows for the specific and efficient in vivo transfection of different regions of the C57BL/6 mouse central nervous system. We also show that histogram analysis is a valuable tool for objectifying and accelerating postmitotic neural migration assays. COMPARISON WITH EXISTING METHODS: Until now, conditions for in utero electroporation of C57BL/6 mice are sparsely defined. Further, compared with time-consuming cell body counting histogram analysis allows objectified and accelerated postmitotic neural migration assays. CONCLUSION: Together, our results provide a manual for the in utero electroporation of specific regions of the central nervous systems C57BL/6 mice and objectified data analysis.
BACKGROUND: In utero electroporation is a fast an efficient tool to specifically address gene expression in the murine central nervous system. This technique was originally established in ICR/CD-1 outbred mice. Neuroanatomical differences between the different mouse strains and variations in gestation length require the optimization of the conditions for each strain to avoid severe complications. Furthermore the relevant position information is currently only scarcely standardized and not always easy to transfer to C57BL/6 mice. NEW METHOD: In this study we present an improved method for in utero electroporation of C57BL/6 including a detailed atlas that allows for specific and efficient in vivo transfection. Further we introduce histogram analysis as a tool for neural migration assays. RESULTS: We report individually adapted conditions for in utero electroporation in C57BL/6 mice that differ from the previously published data for ICR/CD-1 mice. Furthermore, this article outlines a detailed angle-map that allows for the specific and efficient in vivo transfection of different regions of the C57BL/6 mouse central nervous system. We also show that histogram analysis is a valuable tool for objectifying and accelerating postmitotic neural migration assays. COMPARISON WITH EXISTING METHODS: Until now, conditions for in utero electroporation of C57BL/6 mice are sparsely defined. Further, compared with time-consuming cell body counting histogram analysis allows objectified and accelerated postmitotic neural migration assays. CONCLUSION: Together, our results provide a manual for the in utero electroporation of specific regions of the central nervous systems C57BL/6 mice and objectified data analysis.
Authors: Joachim Ahlbeck; Lingzhen Song; Mattia Chini; Sebastian H Bitzenhofer; Ileana L Hanganu-Opatz Journal: Elife Date: 2018-04-10 Impact factor: 8.140