Newly established cell lines fromDrosophila larval CNS express neural specific characteristics

Ui, Kumiko; Nishihara, Shoko; Sakuma, Mayumi; Togashi, Shin; Ueda, Ryo; Miyata, Yuhei; Miyake, Tadashi

doi:10.1007/bf02632042

Cited by 92 publications

(71 citation statements)

References 34 publications

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“…These cells are thought to be derived from embryonic hemocytes, although the exact origin is unknown. BG2 cells are thought to be cells of neuronal origin (Ui et al, 1994). Both types of cells are able to attach and spread on the substratum.…”

Section: Resultsmentioning

confidence: 99%

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Cascade pathway of filopodia formation downstream of SCAR

Biyasheva

Svitkina

Kunda

et al. 2004

Journal of Cell Science

105

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The protrusion of two distinct actin-containing organelles, lamellipodia and filopodia, is thought to be regulated by two parallel pathways: from Rac1 through Scar/WAVEs to lamellipodia, and from Cdc42 through N-WASP to filopodia. We tested this hypothesis in Drosophila, which contains a single gene for each WASP subfamilies, SCAR and WASp. We performed targeted depletion of SCAR or WASp by dsRNA-mediated interference in two Drosophila cultured cell lines expressing lamellipodial and filopodial protrusion. Knockdown was verified by laser capture microdissection and RT-PCR, as well as western blotting. Morphometrical, kinetic and electron microscopy analyses of the SCAR-depleted phenotype in both cell types revealed strong inhibition of lamellipodial formation and cell spreading, as expected. More importantly, filopodia formation was also strongly inhibited, which is not consistent with the parallel pathway hypothesis. By contrast, depletion of WASp did not produce any significant phenotype, except for a slight inhibition of spreading, showing that both lamellipodia and filopodia in Drosophila cells are regulated predominantly by SCAR. We propose a new, cascade pathway model of filopodia regulation in which SCAR signals to lamellipodia and then filopodia arise from lamellipodia in response to additional signal(s).

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Section: Resultsmentioning

confidence: 99%

“…Cell culture and RNAi Drosophila permanent cell lines, S2R+ and Dm-BG2-c2 (referred to as BG2 throughout the paper), and culture conditions have been described previously (Ui et al, 1994;Yanagawa et al, 1998). Cells were split every 4-5 days.…”

Section: Methodsmentioning

confidence: 99%

Cascade pathway of filopodia formation downstream of SCAR

Biyasheva

Svitkina

Kunda

et al. 2004

Journal of Cell Science

105

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“…Cell culture: Kc 167 (Echalier and Ohanessian 1969;Perrimon 2007), S2 (Schneider 1972;Perrimon 2007), D (Debec 1978(Debec , 1984Cherbas 2007), DH-33 (Sondermeijer et al 1980;Cherbas 2007), mbn2 (Haars et al 1980;Cherbas 2007), clone 8 (Peel et al 1990;Perrimon 2007), and MLDmBG2-c6 (Ui et al 1994;Cherbas 2007) cells were grown at 25°following standard protocols. Kc 167 and S2 cultures were grown in sterile filtered Schneider's medium ½GIBCO (Grand Island, NY) no.…”

Section: Methodsmentioning

confidence: 99%

Disruption of Topoisomerase II Perturbs Pairing in Drosophila Cell Culture

Williams¹,

Bateman²,

Novikov³

et al. 2007

Genetics

111

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Homolog pairing refers to the alignment and physical apposition of homologous chromosomal segments. Although commonly observed during meiosis, homolog pairing also occurs in nonmeiotic cells of several organisms, including humans and Drosophila. The mechanism underlying nonmeiotic pairing, however, remains largely unknown. Here, we explore the use of established Drosophila cell lines for the analysis of pairing in somatic cells. Using fluorescent in situ hybridization (FISH), we assayed pairing at nine regions scattered throughout the genome of Kc 167 cells, observing high levels of homolog pairing at all six euchromatic regions assayed and variably lower levels in regions in or near centromeric heterochromatin. We have also observed extensive pairing in six additional cell lines representing different tissues of origin, different ploidies, and two different species, demonstrating homolog pairing in cell culture to be impervious to cell type or culture history. Furthermore, by sorting Kc 167 cells into G 1 , S, and G 2 subpopulations, we show that even progression through these stages of the cell cycle does not significantly change pairing levels. Finally, our data indicate that disrupting Drosophila topoisomerase II (Top2) gene function with RNAi and chemical inhibitors perturbs homolog pairing, suggesting Top2 to be a gene important for pairing.

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“…Three lines were used to look for differences in cohesin binding patterns. Two lines, the Sg4 subline of Schneider line 2 and the Kc167 subline of Kc cells (Echalier and Ohanessian 1970), are embryonic in origin, and the ML-DmBG3 line (BG3; Ui et al 1994) is derived from third instar central nervous system. The Affymetrix tiled microarray contains some 3×10 6 25-nt oligonucleotide features every 35 bp or so.…”

Section: Nipped-b and Cohesin Colocalize Genome-widementioning

confidence: 99%