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dnmt1xenopus   

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Experiment details for dnmt1

Transient depletion of xDnmt1 leads to premature gene activation in Xenopus embryos.

Transient depletion of xDnmt1 leads to premature gene activation in Xenopus embryos.

Gene Clone Species Stages Anatomy
dnmt1.L laevis unfertilized egg stage oocyte , animal , animal pole
dnmt1.L laevis NF stage 7 animal blastomere , animal
dnmt1.L laevis NF stage 11 involuted dorsal mesoderm , involuted ventral mesoderm
dnmt1.L laevis NF stage 15 neuroectoderm , pre-chordal neural plate , neural fold
dnmt1.L laevis NF stage 20 eye primordium , posterior neural tube , anterior neural tube , neural tube , optic field
dnmt1.L laevis NF stage 23 brain , forebrain , midbrain , hindbrain , spinal cord , [+]
dnmt1.L laevis NF stage 35 and 36 brain , forebrain , midbrain , hindbrain , notochord , [+]

  Expression of xDnmt1 during Xenopusdevelopment. (A) The functional domains of the xDnmt1 protein are shown as shaded and solid boxes. (NLS) Nuclear localization signal; (RF) region involved in targeting of the enzyme to replication foci; (Zn) cystine-rich Zn/DNA-binding motif; (PBHD) polybromo-1 protein homologous domain; I are the conserved motifs of the methyltransferase catalytic domain. The 1.4-kb partial somaticxDnmt1 cDNA (st. 202) is 98% homologous to the oocytexDnmt1 catalytic domain (red). The blue bar indicates the PCR product used to detect xDnmt1 depletion in Figure 3A. (B) Northern blot analysis of xDnmt1 transcripts during embryo development. Ornithine decarboxylase (ODC) is an ubiquitously expressed gene. Stages of development are indicated above the xDnmt1 blot. (C,D) Whole-mount in situ hybridization localizes xDnmt1 transcripts (pink) to the animal pole of albino Xenopus egg (C) and animal pole blastomeres of 64-cell blastula (D). (An) Animal pole; (Veg) vegetal pole. (E) xDnmt1 is not detected in the vegetal hemisphere of 64-cell blastula. (F) After MBT the somatic form of xDnmt1 appears in the deep cells of the dorsal (DM) and ventral (VM) mesoderm of stage 11 gastrula (sagittal section). (G) During neurulation at stage 15 the staining forxDnmt1 transcripts is localized in the eye (e) and brain regions (br) of the prospective head neuroectoderm (pne) and along the edges of the open neural fold (nf). (H,I) A similar pattern is maintained during tailbud stages 20 (F) and 23 (G). (fb) Forebrain; (mb) midbrain; (hb) hindbrain. (J) At stage 35 (tadpole) the methyltransferase probe stains eyes, brain regions, branchial arches (ba), posterior notochord (nch), and the motor neurons (mn). (K) xDnmt1 protein (xDnmt1p) was immunoprecipitated from extracts derived from animal or vegetal halves of 64-cell blastulae and detected by Western blot analysis. Both extracts contain equal amounts of PCNA (bottom).