In Utero Electroporation for Manipulation of Specific Neuronal Populations

Abstract

The complexity of brain functions is supported by the heterogeneity of brain tissue and millisecond-scale information processing. Understanding how complex neural circuits control animal behavior requires the precise manipulation of specific neuronal subtypes at high spatiotemporal resolution. In utero electroporation, when combined with optogenetics, is a powerful method for precisely controlling the activity of specific neurons. Optogenetics allows for the control of cellular membrane potentials through light-sensitive ion channels artificially expressed in the plasma membrane of neurons. Here, we first review the basic mechanisms and characteristics of in utero electroporation. Then, we discuss recent applications of in utero electroporation combined with optogenetics to investigate the functions and characteristics of specific regions, layers, and cell types. These techniques will pave the way for further advances in understanding the complex neuronal and circuit mechanisms that underlie behavioral outputs.

Advanced Techniques Using In Vivo Electroporation to Study the Molecular Mechanisms of Cerebral Development Disorders

“…IUE comes with certain limitations. Primarily, controlling the expression levels of exogenous genes in individual cells using IUE poses significant challenges [ 56 ]. This makes it difficult to perform simple quantitative analysis using reporter gene expression levels.…”

Chemogenetic manipulation of astrocyte activity at the synapse— a gateway to manage brain disease

“…However, LVs have been associated with the risk of mutagenesis since they integrate into the genome. IUE provides yet another alternative to express genes of interest in brain cells and has been successfully implemented in several animal models such as mice, rats, ferrets and cats ( Yamashiro et al, 2022 ; Edwards-Faret et al, 2023 ). As astrocytes start developing in the CNS shortly after birth, PALE, an adaptation of IUE, can be used to specifically target astrocytes ( García-Marqués and López-Mascaraque, 2013 ; Gee et al, 2015 ; Stogsdill et al, 2017 ; Kittock and Pilaz, 2023 ).…”

“…In this technique, plasmids, upon injection into the parenchyma of newborn mouse pups (P0-P1), are delivered into the cells due to the action of electrical impulses which disrupt the cell membrane, allowing the passage of the DNA. Besides being less damaging, since the manipulation is performed early in life, and allowing the transfection of bigger constructs than the viral vector approaches, by precisely controlling the injection site and/or the position of the electrodes, both IUE and PALE allow region specific transgene expression ( Yamashiro et al, 2022 ; Kittock and Pilaz, 2023 ). IUE and PALE have been used to express DREADDs in neurons ( Hurni et al, 2017 ; Muthusamy et al, 2017 ).…”