Deletion of a conserved noncoding sequence in Plzf intron leads to Plzf down-regulation in limb bud and polydactyly in the rat

Authors

  • František Liška,

    Corresponding author
    1. Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Praha, Czech Republic
    • Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Albertov 4, 128 00 Praha 2, Czech Republic
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    • Drs. Liška and Šnajdr contributed equally to this work.

  • Pavel Šnajdr,

    1. Institute of Anatomy, 1st Faculty of Medicine, Charles University in Prague, Unemocnice 3, Praha, Czech Republic
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    • Drs. Liška and Šnajdr contributed equally to this work.

  • Lucie Šedová,

    1. Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Praha, Czech Republic
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  • Ondřej Šeda,

    1. Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Praha, Czech Republic
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  • Blanka Chylíková,

    1. Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Praha, Czech Republic
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  • Petra Slámová,

    1. Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Praha, Czech Republic
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  • Eliška Krejčí,

    1. Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Praha, Czech Republic
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  • David Sedmera,

    1. Institute of Anatomy, 1st Faculty of Medicine, Charles University in Prague, Unemocnice 3, Praha, Czech Republic
    2. Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Praha, Czech Republic
    3. Institute of Physiology, Academy of Sciences of the Czech Republic, Praha, Czech Republic
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  • Miloš Grim,

    1. Institute of Anatomy, 1st Faculty of Medicine, Charles University in Prague, Unemocnice 3, Praha, Czech Republic
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  • Drahomíra Křenová,

    1. Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Praha, Czech Republic
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  • Vladimír Křen

    1. Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Praha, Czech Republic
    2. Institute of Physiology, Academy of Sciences of the Czech Republic, Praha, Czech Republic
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Abstract

Lx mutation in SHR.Lx rat manifests in homozygotes as hindlimb preaxial polydactyly. It was previously mapped to a chromosome 8 segment containing the Plzf gene. Plzf (promyelocytic leukemia zinc finger protein) influences limb development as a direct repressor of posterior HoxD genes. However, the Plzf coding sequence is intact in the Lx mutants. Using linkage mapping in F2 hybrids, we downsized the segment containing Lx to 155 kb and sequenced conserved noncoding elements (CNEs) inside. A 2,964-bp deletion in Plzf intron 2, never detected in control animals, is the only candidate for Lx. The deletion removes the most deeply conserved CNE in the 155-kb segment, suggesting a regulatory influence on Plzf expression. Correspondingly, using in situ hybridization and quantitative real-time polymerase chain reaction, we found a decrease of Plzf expression in Lx/Lx limb buds with concomitant anterior expansion of expression domains of its targets, Hoxd1013 genes, in the absence of ectopic Sonic hedgehog expression. Upstream regulation of Plzf in limb buds is currently unknown. We present here the first candidate Plzf cis-regulatory sequence. Developmental Dynamics 238:673–684, 2009. © 2009 Wiley-Liss, Inc.

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