Diaphanous-related formin 2 and profilin I are required for gastrulation cell movements
Resource
PLoS ONE, 3, e3439
Journal
PLoS ONE
Journal Volume
3
Journal Issue
10
Pages
e3439
Date Issued
2008
Date
2008
Author(s)
Lai, Shih-Lei
Chan, Tun-Hao
Lin, Meng-Ju
Lou, Show-Wan
Heisenberg, Carl-Philipp
Abstract
Intensive cellular movements occur during gastrulation. These cellular movements rely heavily on dynamic actin assembly. Rho with its associated proteins, including the Rho-activated formin, Diaphanous, are key regulators of actin assembly in cellular protrusion and migration. However, the function of Diaphanous in gastrulation cell movements remains unclear. To study the role of Diaphanous in gastrulation, we isolated a partial zebrafish diaphanous-related formin 2 (zdia2) clone with its N-terminal regulatory domains. The GTPase binding domain of zDia2 is highly conserved compared to its mammalian homologues. Using a yeast two-hybrid assay, we showed that zDia2 interacts with constitutively-active RhoA and Cdc42. The zdia2 mRNAs were ubiquitously expressed during early embryonic development in zebrafish as determined by RT-PCR and whole-mount in situ hybridization analyses. Knockdown of zdia2 by antisense morpholino oligonucleotides (MOs) blocked epiboly formation and convergent extension in a dose-dependent manner, whereas ectopic expression of a human mdia gene partially rescued these defects. Time-lapse recording further showed that bleb-like cellular processes of blastoderm marginal deep marginal cells and pseudopod-/filopod-like processes of prechordal plate cells and lateral cells were abolished in the zdia2 morphants. Furthermore, zDia2 acts cell-autonomously since transplanted zdia2-knockdown cells exhibited low protrusive activity with aberrant migration in wild type host embryos. Lastly, co-injection of antisense MOs of zdia2 and zebrafish profilin I (zpfn 1), but not zebrafish profilin II, resulted in a synergistic inhibition of gastrulation cell movements. These results suggest that zDia2 in conjunction with zPfn 1 are required for gastrulation cell movements in zebrafish. © 2008 Lai et al.
Other Subjects
actin; antisense oligonucleotide; cytoskeleton protein; formin 2; guanosine triphosphatase; messenger RNA; profilin; profilin i; protein Cdc42; Rho factor; RhoA guanine nucleotide binding protein; unclassified drug; carrier protein; diaphanous related formin 2 protein, Zebrafish; diaphanous-related formin 2 protein, Zebrafish; profilin; protein Cdc42; RhoA guanine nucleotide binding protein; zebrafish protein; amino terminal sequence; animal cell; article; blastoderm; cell migration; cell motion; controlled study; embryo development; filopodium; gastrulation; genetic conservation; in situ hybridization; inhibition kinetics; mammal; nonhuman; nucleotide sequence; protein assembly; protein binding; protein domain; protein expression; protein function; protein interaction; pseudopodium; reverse transcription polymerase chain reaction; sequence homology; two hybrid system; wild type; zebra fish; animal; animal embryo; human; metabolism; nucleic acid probe; physiology; Danio rerio; Mammalia; Pseudopoda; Animals; Antisense Elements (Genetics); Carrier Proteins; cdc42 GTP-Binding Protein; Cell Movement; Embryo, Nonmammalian; Gastrulation; Humans; Profilins; Protein Binding; rhoA GTP-Binding Protein; Two-Hybrid System Techniques; Zebrafish; Zebrafish Proteins
Type
journal article
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