Transfection into Mouse/Rat Brain by Electroporation

NEPA21

Tweezer Electrodes

Needle/Other Electrodes

Petridish/Microscope Slide Electrodes

Cables, Foot Switch & Others

Transfection into Mouse/Rat Brain by Electroporation

APPLICATIONS

Electroporation-mediated gene transfer in the adult rat brain

Figure A: EGFP expression in the medial preoptic nuclei of a female rat examined 4 days after bilateral electroporation at 10 weeks of age. (An asterisk indicates the trace of the positioning of the electrode)

Figure B: EGFP-positive cells (high magnification of Fig. A using a 60x objective lens). EGFP fluorescent signals are observed in the perikarya.

Figure C: Estrogen receptor αimmunoreactivity in the medial preoptic nuclei and the periventricular nuclei of an adult female rat.3V: third ventricle.

Figure D: Estrogen receptor α-positive cells (high magnification of Fig. C using a 60x objective lens). Estrogen receptor  immunoreactivity is prominent in the nuclei.

Tetsuo Shirakawa, Center for Advanced Oral Medicine, Hokkaido University Hospital

Electroporation-mediated gene transfer system applied to cultured CNS neurons

Schematic representation of an electroporation set-up.

A fragment of the mouse embryonic hippocampus was placed on a Millipore membrane filter and 5μl EP buffer containing 1mg/ml of plasmid DNA was applied onto the tissue.
A tungsten needle was attached to the surface of a droplet.
After application of square pulses the tissue fragment was returned to a petri dish containing ice-cold HBSS solution.

Electroporation-mediated expression of fluorescent proteins in hippocampal neurons.

(a-c) Organ culture of hippocampal tissue fragments three days after electroporation with CAG-eGFP
(a) Ta1X4 -eGFP, (b) and b-actin-eGFP, (c) expression constructs.

(d,e) A mature hippocampal neuron maintained 14 days in dissociated culture after electroporation of a -actin-eGFP expression construct. Higher magnification view of the region marked by a rectangle in (d) reveals dendritic spines on the surface of dendritic shafts (arrows in e).

(f,g) A hippocampal neuron 7 days after electroporation of 1:1 mixture of Ta1X4-eGFP and Ta1X4-mRFP1. Both eGFP fluorescence (f) and mRFP1 fluorescence (g) can be observed in a single cell.

(h) Relative fluorescence intensity of hippocampal tissue fragments after electroporation of eGFP-expression plasmids with four different promoter sequences. The tissue fragments were maintained in culture for 4 days, fixed and observed under a confocal microscope. Fluorescence intensities per unit area of the tissue fragments were determined.

(i) 2Relative fluorescence intensity of hippocampal tissue fragments isolated at two different developmental stages and electroporated with b-actin-eGFP. Tissue fragments were maintained for 4 days in culture and subsequently fixed. Fluorescence intensities were measured using a confocal microscope.

 

 

Shigeo Okabe, Department of Cellular Neurobiology, Graduate School of Medicine and Faculty of Medicine, The University of Tokyo
*Neuroreport, Volume 15, Issue 6, Pages 971-975, April 29, 2004

PUBLICATIONS

  • Adult

Genome editing of Nf1, Pten, and Trp53 in neonatal mice induces glioblastomas positive for oligodendrocyte lineage transcription factor 2

Hiromi Yamamoto, Keisuke Yamamura, Haruka Nagasaki, Takamasa Suzuki, Fumiko Ninomiya, Kenji Matsubara, Naomoto Harada, Shuichi Ohkubo

J Toxicol Pathol. 2021 Oct;34(4):359-365.

  • Adult and New-born

Primary cilium-dependent cAMP/PKA signaling at the centrosome regulates neuronal migration

Julie Stoufflet, Maxime Chaulet, Mohamed Doulazmi, Coralie Fouquet, Caroline Dubacq, Christine Métin, Alain Trembleau, Pierre Vincent, Isabelle Caillé

bioRxiv November 12, 2019

  • Adult

Rho Family GTPases, Rac and Cdc42, Control the Localization of Neonatal Dentate Granule Cells During Brain Development

Hidenori Ito, Rika Morishita, Makoto Mizuno, Hidenori Tabata, Koh-Ichi Nagata

Hippocampus, 29 (7), 569-578 Jul 2019

  • Adult

A Dual Role for the Transcription Factor Sp8 in Postnatal Neurogenesis

Elodie Gaborieau, Anahi Hurtado-Chong, Maria Fernández, Kasum Azim, Olivier Raineteau

Sci Rep, 8 (1), 14560 2018 Sep 28

    Precise Somatotopic Thalamocortical Axon Guidance Depends on LPA-Mediated PRG-2/Radixin Signaling

    Jin Cheng, Sadhna Sahani, Torben Johann Hausrat, Jenq-Wei Yang, Haichao Ji, Nikolai Schmarowski, Heiko Endle, Xinfeng Liu, Yunbo Li, Rahel Böttche, Konstantin Radyushkin, Hans M Maric, Anna Hoerder-Suabedissen, Zoltán Molnár, Pierre-Hugues Prouvot, Thorsten Trimbuch, Olaf Ninnemann, Jisen Huai, Wei Fan, Barbara Visentin, Roger Sabbadini, Kristian Strømgaard, Albrecht Stroh, Heiko J Luhmann, Matthias Kneussel, Robert Nitsch, Johannes Vogt

    Neuron, 92 (1), 126-142 2016 Oct 5

      Cortical excitatory neurons become protected from cell division during neurogenesis in an Rb family-dependent manner.

      Oshikawa M et al.

      Development. 2013 Jun;140(11):2310-20.

        Wnt/calcium signaling mediates axon growth and guidance in the developing corpus callosum.

        Hutchins BI et al.

        Dev Neurobiol. 2011 Apr;71(4):269-83.

        • Adult

        Cellular Mechanismhs Underlying Morphine Analgesic Tolerance and Hyperalgesia

        Hiroshi Ueda

        Opioid-Induced Hyperalgesia, First, Pages 9-20, October 2009

        • New-born

        Efficient In Vivo Electroporation of the Postnatal Rodent Forebrain

        Boutin et al.

        PLoS ONE, Volume 3, Issue 4, e1883, 2 April 2008

        • New-born

        Plexin-A2 and its ligand, Sema6A, control nucleus-centrosome coupling in migrating granule cells

        Renaud et al.

        Nature Neuroscience, Volume 11, Number 4, Pages 440-449, April 2008

        • Adult

        Genetic manipulation of adult mouse neurogenic niches by in vivo electroporation

        Barnabe-Heider et al.

        Nature Methods, Volume 5, Number 2, Pages 189-196, February 2008

        • New-born

        Microtubule-based nuclear movement occursindependently of centrosome positioning in migrating neurons

        Umeshima et al.

        Proc Natl Acad Sci U S A . 2007 Oct 9;104(41):16182-7.

          Homer proteins control neuronal differentiation through IP(3) receptor signaling.

          Tanaka M et al.

          FEBS Lett. 2006 Nov 13;580(26):6145-50.

            The Caudal Migratory Stream: A Novel Migratory Stream of Interneurons Derived from the Caudal Ganglionic Eminence in the Developing Mouse Forebrain

            Yozu M, Tabata H, Nakajima K.

            J Neurosci. 2005 Aug 3;25(31):7268-77.

              Electroporation-mediated gene transfer system applied to cultured CNS neurons

              Kawabata I, Umeda T, Yamamoto K, Okabe S.

              Neuroreport . 2004 Apr 29;15(6):971-5.

              • Adult

              Locus-Specific Rescue of GluR1 NMDA Receptors in Mutant Mice Identifies the Brain Regions Important for Morphine Tolerance and Dependence

              Inoue et al.

              The Journal of Neuroscience, Volume 23, Issue 16, Pages 6529-6536, 23 July 2003

              • Adult

              Up-regulation of Protein-disulfide Isomerase in Response to Hypoxia/Brain Ischemia and Its Protective Effect against Apoptotic Cell Death

              Tanaka et al.

              J Biol Chem . 2000 Apr 7;275(14):10388-93.

              Electroporation

              Drug Delivery and Transfection

              Electro Cell Fusion

              Fluorescent Staining

              Single-Cell/Micro-Particle Transfer

              Cell Freezing

              Mechanical Vibration