Targeted electroporation in Xenopus tadpoles in vivo -: from single cells to the entire brain

被引:107
作者
Haas, K [1 ]
Jensen, K [1 ]
Sin, WC [1 ]
Foa, L [1 ]
Cline, HT [1 ]
机构
[1] Cold Spring Harbor Lab, Cold Spring Harbor, NY 11724 USA
关键词
electroporation; synaptic plasticity; transfection; functional genomics; neuronal imaging;
D O I
10.1046/j.1432-0436.2002.700404.x
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Electroporation is becoming more popular as a technique for transfecting neurons within intact tissues. One of the advantages of electroporation over other transfection techniques is the ability to precisely target an area for transfection. Here we highlight this advantage by describing methods to restrict transfection to either a single cell, clusters of cells, or to include large portions of the brain of the intact Xenopus tadpole. Electroporation is also an effective means of gene delivery in the retina. We have developed these techniques to examine the effects of regulated gene expression on various neuronal properties, including structural plasticity and synaptic transmission. Restriction of transfection to individual cells aids in imaging of neuronal morphology, while bulk cell transfection allows examination of the affects of gene expression on populations of cells by biochemical assays, imaging, and electrophysiological recording.
引用
收藏
页码:148 / 154
页数:7
相关论文
共 26 条
[1]   Mammalian ELAV-like neuronal RNA-binding proteins HuB and HuC promote neuronal development in both the central and the peripheral nervous systems [J].
Akamatsu, W ;
Okano, HJ ;
Osumi, N ;
Inoue, T ;
Nakamura, S ;
Sakakibara, SI ;
Miura, M ;
Matsuo, N ;
Darnell, RB ;
Okano, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (17) :9885-9890
[2]  
Araki I, 1999, DEVELOPMENT, V126, P5127
[3]   Localized electroporation: A method for targeting expression of genes in avian embryos [J].
Atkins, RL ;
Wang, D ;
Burke, RD .
BIOTECHNIQUES, 2000, 28 (01) :94-+
[4]   A homeodomain protein code specifies progenitor cell identity and neuronal fate in the ventral neural tube [J].
Briscoe, J ;
Pierani, A ;
Jessell, TM ;
Ericson, J .
CELL, 2000, 101 (04) :435-445
[5]  
Buono Russel J., 1992, Molecular Marine Biology and Biotechnology, V1, P271
[6]   Electroporation-mediated gene transfer in free-swimming embryonic Xenopus laevis [J].
Eide, FF ;
Eisenberg, SR ;
Sanders, TA .
FEBS LETTERS, 2000, 486 (01) :29-32
[7]   The scaffold protein, Homer1b/c, regulates axon pathfinding in the central nervous system in vivo [J].
Foa, L ;
Rajan, I ;
Haas, K ;
Wu, GY ;
Brakeman, P ;
Worley, P ;
Cline, H .
NATURE NEUROSCIENCE, 2001, 4 (05) :499-506
[8]   Single-cell electroporation for gene transfer in vivo [J].
Haas, K ;
Sin, WC ;
Javaherian, A ;
Li, Z ;
Cline, HT .
NEURON, 2001, 29 (03) :583-591
[9]  
Inoue T, 2001, DEVELOPMENT, V128, P561
[10]   An impulse to the brain -: using in vivo electroporation [J].
Inoue, T ;
Krumlauf, R .
NATURE NEUROSCIENCE, 2001, 4 (Suppl 11) :1156-1158