In Utero Electroporation of the Mouse Embryo

Saito, Tetsuichiro

doi:10.1007/978-1-4939-2459-2_1

Cited by 5 publications

(2 citation statements)

References 59 publications

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“…The in utero electroporation of Sema5a siRNA into the embryonic mouse brain was performed as described previously using C57BL/6J strain mice with slight modifications [ 79 ]. Animal experiments were conducted following the guidelines the National Institutes of Health set forth as outlined in the Guide for the Care and Use of Laboratory Animals.…”

Section: Methodsmentioning

confidence: 99%

Sex-specific impacts of prenatal bisphenol A exposure on genes associated with cortical development, social behaviors, and autism in the offspring’s prefrontal cortex

Kanlayaprasit,

Saeliw,

Thongkorn

et al. 2024

Biol Sex Differ

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Background Recent studies have shown that prenatal BPA exposure altered the transcriptome profiles of autism-related genes in the offspring’s hippocampus, disrupting hippocampal neuritogenesis and causing male-specific deficits in learning. However, the sex differences in the effects of prenatal BPA exposure on the developing prefrontal cortex, which is another brain region highly implicated in autism spectrum disorder (ASD), have not been investigated. Methods We obtained transcriptome data from RNA sequencing analysis of the prefrontal cortex of male and female rat pups prenatally exposed to BPA or control and reanalyzed. BPA-responsive genes associated with cortical development and social behaviors were selected for confirmation by qRT-PCR analysis. Neuritogenesis of primary cells from the prefrontal cortex of pups prenatally exposed to BPA or control was examined. The social behaviors of the pups were assessed using the two-trial and three-chamber tests. The male-specific impact of the downregulation of a selected BPA-responsive gene (i.e., Sema5a) on cortical development in vivo was interrogated using siRNA-mediated knockdown by an in utero electroporation technique. Results Genes disrupted by prenatal BPA exposure were associated with ASD and showed sex-specific dysregulation. Sema5a and Slc9a9, which were involved in neuritogenesis and social behaviors, were downregulated only in males, while Anxa2 and Junb, which were also linked to neuritogenesis and social behaviors, were suppressed only in females. Neuritogenesis was increased in males and showed a strong inverse correlation with Sema5a and Slc9a9 expression levels, whereas, in the females, neuritogenesis was decreased and correlated with Anxa2 and Junb levels. The siRNA-mediated knockdown of Sema5a in males also impaired cortical development in utero. Consistent with Anxa2 and Junb downregulations, deficits in social novelty were observed only in female offspring but not in males. Conclusion This is the first study to show that prenatal BPA exposure dysregulated the expression of ASD-related genes and functions, including cortical neuritogenesis and development and social behaviors, in a sex-dependent manner. Our findings suggest that, besides the hippocampus, BPA could also exert its adverse effects through sex-specific molecular mechanisms in the offspring’s prefrontal cortex, which in turn would lead to sex differences in ASD-related neuropathology and clinical manifestations, which deserves further investigation.

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Section: Methodsmentioning

confidence: 99%

Sex-specific impacts of prenatal bisphenol A exposure on genes associated with cortical development, social behaviors, and autism in the offspring’s prefrontal cortex

Kanlayaprasit,

Saeliw,

Thongkorn

et al. 2024

Biol Sex Differ

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“…Plasmids expressing NeuroD-CRE-IRES-GFP or NeuroD-X-IRES-GFP (where X is the gene of interest) at 3 μg/μl. E14.5 floxed embryos or wild-type embryos Reagents and equipment for in utero electroporation (see Pacary and Guillemot, 2014;Saito, 2015) Day 1: In utero electroporation 1. Perform in utero electroporation as previously described (Pacary and Guillemot, 2014;Saito, 2015).…”

Section: Additional Materials (See Also Basic Protocol 1)mentioning

confidence: 99%

Real‐time Recordings of Migrating Cortical Neurons from GFP and Cre Recombinase Expressing Mice

2016

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The cerebral cortex is one of the most intricate regions of the brain that requires elaborate cell migration patterns for its development. Experimental observations show that projection neurons migrate radially within the cortical wall, whereas interneurons migrate along multiple tangential paths to reach the developing cortex. Tight regulation of the cell migration processes ensures proper positioning and functional integration of neurons to specific cerebral cortical circuits. Disruption of neuronal migration often leads to cortical dysfunction and/or malformation associated with neurological disorders. Unveiling the molecular control of neuron migration is thus fundamental to understanding the physiological or pathological development of the cerebral cortex. In this unit, protocols allowing detailed analysis of patterns of migration of both interneurons and projection neurons under different experimental conditions (i.e., loss or gain of function) are presented.

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Control of Gene Expression in Neurons by the Use of In Vivo Electroporation and the Tetracycline System

Sato

Muroyama

Saito

2015

Electroporation Methods in Neuroscience

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In Utero Electroporation of the Mouse Embryo

Cited by 5 publications

References 59 publications

Sex-specific impacts of prenatal bisphenol A exposure on genes associated with cortical development, social behaviors, and autism in the offspring’s prefrontal cortex

Sex-specific impacts of prenatal bisphenol A exposure on genes associated with cortical development, social behaviors, and autism in the offspring’s prefrontal cortex

Real‐time Recordings of Migrating Cortical Neurons from GFP and Cre Recombinase Expressing Mice

Control of Gene Expression in Neurons by the Use of In Vivo Electroporation and the Tetracycline System

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