Misexpression of genes in brain vesicles by <i>in ovo</i> electroporation

Nakamura, Harukazu; Watanabe, Yuji; Funahashi, Junichi

doi:10.1046/j.1440-169x.2000.00501.x

Cited by 71 publications

(47 citation statements)

References 12 publications

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“…We electroporated chick embryos, as described by Nakamura et al (2000). Briefly, this involved injecting 1 mg/ml plasmid DNA into the closed neural tube at the eight-to ten-somite stage (Hamburger and Hamilton [HH] stage 9 -10; 33-36 hr), and applying five electric pulses (20-mV square pulses at 50-ms intervals) through electrodes spaced 4 mm apart; we used an Electrosquareporater T820 (BTX, Genetronics) during this process.…”

Section: Electroporationmentioning

confidence: 99%

Conserved RARE localization in amphioxusHox clusters and implications forHox code evolution in the vertebrate neural crest

et al. 2006

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Section: Electroporationmentioning

confidence: 99%

Conserved RARE localization in amphioxusHox clusters and implications forHox code evolution in the vertebrate neural crest

et al. 2006

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“…However Muramatsu et al (1997) showed that gene transfer could be obtained by in ovo electroporation. We set the precise conditions of electroporation to misexpress a gene of interest in the neural tube of chick embryos (Funahashi et al, 1999;Nakamura et al, 2000;Nakamrua and Funahashi, 2001). This method is so effective that chick embryos were revivified as experimental material for developmental biology.…”

Section: Gene Transfer and Silencing By In Ovo Electroporationmentioning

confidence: 99%

Isthmus organizer and regionalization of the mesencephalon and metencephalon

Nakamura

Watanabe²

2005

Int. J. Dev. Biol.

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The brain vesicles that are formed at an early stage of neural development are the fundamentals of the brain plan. Heterotopic transplantation revealed that the diencephalon could change its fate when juxtaposed to the isthmus (mes-metencephalic boundary), which indicated that the isthmus functions as an organizer for the mesencephalon and metencephalon. Fgf8 is identified as an isthmus organizing signal. Misexpression of Fgf8a and Fgf8b indicated that a strong Fgf8 signal organizes cerebellar development. The transcription factors define the fate of the region. Overlapping expression of Otx2, En1 and Pax2 may define the mesencephalic region and additional expression of Pax3/7 may instruct the mesencephalic region to differentiate into the tectum. The di-mesencephalic boundary is determined by repressive interaction between Pax6 and En1/Pax2 and the mes-metencephalic boundary is defined by repressive interaction between Otx2 and Gbx2. Fgf8 is induced at the border of the Otx2 and Gbx2 expression domain, overlapping with Gbx2 expression.

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“…Since then, many functional studies based on electroporation-mediated gene transfer in chick have been published. Most of this work has been done in neural tissue (e.g., Araki and Nakamura, 1999;Megason and McMahon, 2002), but also in head ectoderm (Ogino and Yasuda, 1998), limb mesenchyme Swartz et al, 2001a,b;Oberg et al, 2002), the segmental plate (Dubrulle et al, 2001), and other tissues (reviewed in Itasaki et al, 1999;Swartz et al, 2001a; for technical overviews see Yasuda et al, 2000;Nakamura et al, 2000;Nakamura and Funahashi, 2001). Recent reports on gene silencing by electroporation of dsRNA in the neural tube (Pekarik et al, 2003) and the use of photoactivatable green fluorescent protein (GFP) for selective labelling of cells and subcellular structures (Patterson and Lippincott-Schwartz, 2002) are further landmarks indicating the immense potential of electroporation to decipher the cellular and molecular mechanisms regulating vertebrate development.…”

Section: Introductionmentioning

confidence: 99%