Project description:The response of ectodermal explants, neuralized by noggin and treated with cycloheximide, following activation of hormone-inducible zic1 injected into the parent embryos compared to those from beta globin injected embryos as controls, is expected to provide information on the direct targets of the Zic1 transcription factor. Experiment Overall Design: Activation of zic1 in ectodermal explants following inhibition of new protein synthesis allowed the direct targets of zic1 to be identified by comparison with controls. After RNA extraction, purification and checks with PCR with actin primers for any mesoderm contamination samples were prepared for hybridization to Xenopus laevis Affymetrix GeneChip arrays.
Project description:The response of ectodermal explants, neuralized by noggin and treated with cycloheximide, following activation of hormone-inducible zic1 injected into the parent embryos compared to those from beta globin injected embryos as controls, is expected to provide information on the direct targets of the Zic1 transcription factor.
Project description:The Xenopus POU class V transcription factor XOct-25 has been shown to inhibit BMP-dependent epidermal differentiation and promote neural induction in the ectoderm during early embryogenesis. In order to identify genes that act downstream of XOct-25, transcriptional profile of Xenopus ectodermal cells overexpressing XOct-25 was compared with control cells by using a DNA microarray method. Two independent experiments. Each experiment contains ectodermal cells overexpressing XOct-25 and corresponding control cells. Xenopus embryos were injected at the 8-cell stage with mRNA encoding GR-XOct-25, a hormone-inducible version of XOct-25. Explants from stage 9 embryos were treated with or without dexamethazone (DEX) until the sibling embryos reached stage 10.5, when they were used for RNA extraction. The explants cultured without DEX was used as a control sample. biological replicates: Sample name XOct-exp 1, Sample name XOct-exp 2
Project description:Pax3 and Zic1 trigger the early neural crest gene regulatory network by the direct activation of multiple key neural crest specifiers [Xenopus_laevis]
Project description:Pax3 and Zic1 trigger the early neural crest gene regulatory network by the direct activation of multiple key neural crest specifiers [X_laevis_2]
Project description:The transcription factors Pax3 and Zic1 are among the earliest genes activated at the neural plate border. Pax3 and Zic1 in combination promote neural crest fate, while Zic1 alone regulate cranial placode progenitor formation. We used microarrays to identify the global repertoire of genes activated by these facors individually or in combination to gain insights into the molecular mechanisms underlying cell fate decision at the neural plate border. Xenopus laevis embryos were injected at the 2-cell stage with mRNA encoding Pax3GR and Zic1GR , the hormone-inducible version of Pax3 and Zic1, alone or in combination. At the blastula stage (stage 9), animal cap explants were dissected and cultured for 8 hours in the presence of dexamethasone. A glucocorticoid receptor (GR) mRNA was also injected as negative control.