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Experiment details for egr2



Knockdown of the complete Hox paralogous group 1 leads to dramatic hindbrain and neural crest defects.

Gene Clone Species Stages Anatomy
egr2.L laevis NF stage 24 hindbrain , cranial neural crest , rhombomere R3 , rhombomere R5

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  Fig. 2. Specificity of PG1 morpholinos. Two morpholinos for each Hox PG1 gene were injected into the left-hand side of the embryo and Krox20 (r3 and r5) and Engrailed-2 (midbrain/hindbrain boundary: * in A-I) expression analysed. The hindbrain region of early tailbud stage embryos (anterior to the top) is shown for non injected controls (NIC; A,D,G), Hoxa1 1st (B: 80%, n=15) and 2nd (C: 70%, n=10) morpholinos, Hoxb1 1st (E: 85%, n=13) and 2nd (F: 70%, n=20) morpholinos and Hoxd1 1st (H: 90%, n=11) and 2nd (I: 80%, n=15) morpholinos. Overexpression of Hoxd1 morpholino insensitive RNA (J) and rescue of Hoxd1 1st and 2nd morpholinos with Hoxd1 RNA is also shown (K,L). Dotted line indicates the midline.

Gene Clone Species Stages Anatomy
egr2.L laevis NF stage 24 hindbrain , cranial neural crest , rhombomere R3 , rhombomere R5 , anterior branchial crest

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  Fig. 5. Triple PG1 knockdown severely affects hindbrain patterning. Morpholinos for each of the Hox PG1 genes (A1/B1/D1MO) were co-injected into the lhs of the embryo (B,D,F,H,J). Non-injected controls are also shown (A,C,E,G,I). The hindbrain region is shown, with anterior to the top. Expression of the neural marker Nrp1 (A,B: 100%, n=11), the anterior marker Otx2 (C,D: 100%, n=10), Gbx2, which is expressed in r1 (E,F: 77%, n=13) and Krox20 (G,H: no stripes 41%; one faint stripe 35%, n=71), was analysed. Dotted line indicates the midline. In later embryos (st. 46), the neural antibody 2-G9 was used to show the morphology of the hindbrain [NIC, I; A1/B1/D1MO, injected on the left-hand side (lhs) *, J: 100%, n=5]. Rhombomere boundaries are marked.

Gene Clone Species Stages Anatomy
egr2.L laevis NF stage 26 hindbrain , cranial neural crest , rhombomere R3 , rhombomere R5 , anterior branchial crest

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  Fig. 3. The triple PG1 knockdown phenotype is more severe than the double knockdowns. Morpholinos for each Hox PG1 gene were injected either in double (D: A1/B1 52%, n=25; E: B1/D1 48%, n=29; F: D1/A1 64%, n=33) or triple (C: A1/B1/D1 94%, n=35) combinations into the left-hand side of the embryo and Krox20 and Engrailed-2 (*) expression analysed. The hindbrain region of early tailbud stage embryos is shown, with anterior to the top. Non-injected controls (NIC; A), and embryos injected on the left-hand side of the embryo with control morpholino (B) are also shown. Arrowheads indicate neural crest expression.

Gene Clone Species Stages Anatomy
egr2.L laevis NF stage 27 hindbrain , cranial neural crest , rhombomere R3 , rhombomere R5 , anterior branchial crest

  Fig. 4. Hoxd1 mRNA can partially rescue the triple PG1 morpholino phenotype. Early tailbud stage embryos were injected on the left-hand side (lhs) with all three PG1 morpholinos (A1/B1/D1 MO) either alone (B,C), or in combination with Hoxd1 mRNA (E,F). Non-injected controls (NIC) and control morpholino (Cont MO) are also shown (A,D). Anterior is to the top. G: Krox-20 expression is partially rescued by Hoxd1 RNA. Numbers of embryos with two stripes (E), one broad stripe (F), one narrow stripe (B) or no stripes (C) of Krox20 expression were scored. Co-injection of Hoxd1 RNA partially rescued the Krox20 expression (n=29), which was either absent or very much reduced in embryos injected only with A1/B1/D1MO (n=18).