title: Targeted gene delivery in the cricket brain, using in vivo electroporation.
abstract: The cricket (Gryllus bimaculatus) is a hemimetabolous insect that is emerging as a model organism for the study of neural and molecular mechanisms of behavioral traits. However, research strategies have been limited by a lack of genetic manipulation techniques that target the nervous system of the cricket. The development of a new method for efficient gene delivery into cricket brains, using in vivo electroporation, is described here. Plasmid DNA, which contained an enhanced green fluorescent protein (eGFP) gene, under the control of a G. bimaculatus actin (Gb'-act) promoter, was injected into adult cricket brains. Injection was followed by electroporation at a sufficient voltage. Expression of eGFP was observed within the brain tissue. Localized gene expression, targeted to specific regions of the brain, was also achieved using a combination of local DNA injection and fine arrangement of the electroporation electrodes. Further studies using this technique will lead to a better understanding of the neural and molecular mechanisms that underlie cricket behaviors.
authors.affiliation: Department of Biological Science, Faculty of Science, Hokkaido University, Kita 10 Nishi 8, Kita-ku, Sapporo 060-0810, Japan.
authors.forename: Chihiro Sato
authors.initials: CS
authors.lastname: Matsumoto
authors.forename: Hisashi
authors.initials: H
authors.lastname: Shidara
authors.forename: Koji
authors.initials: K
authors.lastname: Matsuda
authors.forename: Taro
authors.initials: T
authors.lastname: Nakamura
authors.forename: Taro
authors.initials: T
authors.lastname: Mito
authors.forename: Yukihisa
authors.initials: Y
authors.lastname: Matsumoto
authors.forename: Kotaro
authors.initials: K
authors.lastname: Oka
authors.forename: Hiroto
authors.initials: H
authors.lastname: Ogawa
mesh_terms: D000818:Animals; D001921:Brain; D018274:Electroporation; D018014:Gene Transfer Techniques; D006135:Gryllidae; D008297:Male
keywords: Brain; Cricket; Electroporation; Green fluorescent protein; Insect; Neuron; PP; SR; TP; eGFP; enhanced green fluorescent protein; poring pulse; success rate; transfer pulse