Abstract Objective To establish a system for regulating the gene expression of embryonic mouse cerebral cortex neural stem cells (NSCs) using in utero electroporation (IUE). Methods At embryonic day 14.5, the mouse cerebral cortex NSCs were electro-transfected with the pCIG plasmid injected into the ventricle of the mouse embryo. At embryonic day 16.5 or day 17.5, embryonic mouse brain tissues were collected to prepare frozen sections. Immunofluorescence staining was used to observe the proliferation, apoptosis, division, directional differentiation, migration, and maturation of NSCs. Results The differentiation of NSCs into intermediate progenitors, the proliferation and apoptosis of NSCs, and the morphological development of radial axis of radial glial cells were observed at embryonic day 16.5. The differentiation of NSCs into neurons in layers V-VI of the cerebral cortex, the migration of NSCs to the lateral cerebral cortex, the development of dendrites of migrating neurons, and the maturation of neurons were observed at embryonic day 17.5. Conclusions The system for regulating the gene expression of embryonic mouse cerebral cortex NSCs can be established using IUE, which is useful for the study of neural development related to the proliferation, apoptosis, division, directional differentiation, migration and maturation of NSCs in the cerebral cortex. Citation:Chinese Journal of Contemporary Pediatrics, 2022, 24(9): 1061-1067
OU Wei-Ming,HE Long-Kai,WANG Xiao-Yu et al. Establishment of a system for regulating the gene expression of embryonic mouse cerebral cortex neural stem cells by in utero electroporation[J]. CJCP, 2022, 24(9): 1061-1067.
OU Wei-Ming,HE Long-Kai,WANG Xiao-Yu et al. Establishment of a system for regulating the gene expression of embryonic mouse cerebral cortex neural stem cells by in utero electroporation[J]. CJCP, 2022, 24(9): 1061-1067.
Lindsey BW, Hall ZJ, Heuzé A, et al. The role of neuro-epithelial-like and radial-glial stem and progenitor cells in development, plasticity, and repair[J]. Prog Neurobiol, 2018, 170: 99-114. PMID: 29902500. DOI: 10.1016/j.pneurobio.2018.06.004.
Hoerder-Suabedissen A, Molnár Z. Development, evolution and pathology of neocortical subplate neurons[J]. Nat Rev Neurosci, 2015, 16(3): 133-146. PMID: 25697157. DOI: 10.1038/nrn3915.
Oliveira NCM, Lins éM, Massirer KB, et al. Translational control during mammalian neocortex development and postembryonic neuronal function[J]. Semin Cell Dev Biol, 2021, 114: 36-46. PMID: 33020045. DOI: 10.1016/j.semcdb.2020.09.006.
Dhaya I, Griton M, Konsman JP. Magnetic resonance imaging under isoflurane anesthesia alters cortical cyclooxygenase-2 expression and glial cell morphology during sepsis-associated neurological dysfunction in rats[J]. Animal Model Exp Med, 2021, 4(3): 249-260. PMID: 34557651. PMCID: PMC8446714. DOI: 10.1002/ame2.12167.
Sato M, Takabayashi S, Akasaka E, et al. Recent advances and future perspectives of in vivo targeted delivery of genome-editing reagents to germ cells, embryos, and fetuses in mice[J]. Cells, 2020, 9(4): 799. PMID: 32225003. PMCID: PMC7226049. DOI: 10.3390/cells9040799.
Taniguchi Y, Young-Pearse T, Sawa A, et al. In utero electroporation as a tool for genetic manipulation in vivo to study psychiatric disorders: from genes to circuits and behaviors[J]. Neuroscientist, 2012, 18(2): 169-179. PMID: 21551077. PMCID: PMC3530425. DOI: 10.1177/1073858411399925.
Baumgart J, Grebe N. C57BL/6-specific conditions for efficient in utero electroporation of the central nervous system[J]. J Neurosci Methods, 2015, 240: 116-124. PMID: 25445056. DOI: 10.1016/j.jneumeth.2014.11.004.
Comer AL, Sriram B, Yen WW, et al. A pipeline using bilateral in utero electroporation to interrogate genetic influences on rodent behavior[J]. J Vis Exp, 2020(159): e61350. PMID: 32510510. DOI: 10.3791/61350.
Nishimura YV, Shinoda T, Inaguma Y, et al. Application of in utero electroporation and live imaging in the analyses of neuronal migration during mouse brain development[J]. Med Mol Morphol, 2012, 45(1): 1-6. PMID: 22431177. DOI: 10.1007/s00795-011-0557-0.
