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1.
Dev Biol ; 229(1): 128-40, 2001 Jan 01.
Article in English | MEDLINE | ID: mdl-11133159

ABSTRACT

The Pax family of transcription factors plays important roles in vertebrate organogenesis. Pax-2 is a critical factor in the development of the mammalian urogenital system. Pax-2 is expressed in the epithelia of the ureter, the Müllerian duct, and the Wolffian duct and in the nephrogenic mesenchyme. Gene targeting in the mouse as well as natural mutations in mouse and man have demonstrated the requirement of Pax-2 in the development of these structures. Little is known about the molecular mechanisms regulating Pax-2 expression in the developing urogenital system. As a first step to reveal these mechanisms and to search for the elements and factors controlling Pax-2 expression we have characterized regulatory sequences of the Pax-2 gene in an in vivo reporter assay in the mouse. An 8.5-kb genomic region upstream of the Pax-2 transcription start site directed reporter gene activity in the epithelium of the pronephric duct at 8.25 days postcoitum (dpc) and in the Wolffian duct starting from 9.0 dpc. Expression in the Wolffian duct and its derivatives, the ureter, the collecting duct system, the seminal vesicles, the vas deferens, and the epididymis, was maintained at least until 18.5 dpc. Hence, an element(s) in the 8.5-kb upstream region is sufficient to initiate and maintain Pax-2 expression in the Wolffian duct and its derivatives. In order to more precisely map the Wolffian duct regulatory sequences, a deletion analysis of the 8.5-kb upstream region was performed in a transient in vivo reporter assay. A 0.4-kb subfragment was required for marker gene expression in the Wolffian duct. Misexpression of fgf8 under the control of the 8.5-kb upstream region resulted in polycystic kidneys, demonstrating the general usefulness of Pax-2 regulatory sequences in misexpression of foreign genes in the ureter and collecting duct system of the kidney in transgenic approaches in mice.


Subject(s)
DNA-Binding Proteins/genetics , Regulatory Sequences, Nucleic Acid , Transcription Factors/genetics , Urogenital System/embryology , Wolffian Ducts/embryology , Animals , Base Sequence , Epithelium/embryology , Gene Expression Regulation, Developmental , Genes, Reporter , Genotype , In Situ Hybridization , Mice , Mice, Mutant Strains , Molecular Sequence Data , PAX2 Transcription Factor , Transgenes , Ureter/embryology
2.
Development ; 125(14): 2587-97, 1998 Jul.
Article in English | MEDLINE | ID: mdl-9636074

ABSTRACT

In Hoxa-2(-/- )embryos, the normal skeletal elements of the second branchial arch are replaced by a duplicated set of first arch elements. We show here that Hoxa-2 directs proper skeletal formation in the second arch by preventing chondrogenesis and intramembranous ossification. In normal embryos, Hoxa-2 is expressed throughout the second arch mesenchyme, but is excluded from the chondrogenic condensations. In the absence of Hoxa-2, chondrogenesis is activated ectopically within the rostral Hoxa-2 expression domain to form the mutant set of cartilages. In Hoxa-2(-/- )embryos the Sox9 expression domain is shifted into the normal Hoxa-2 domain. Misexpression of Sox9 in this area produces a phenotype resembling that of the Hoxa-2 mutants. These results indicate that Hoxa-2 acts at early stages of the chondrogenic pathway, upstream of Sox9 induction. We also show that Hoxa-2 inhibits dermal bone formation when misexpressed in its precursors. Furthermore, molecular analyses indicate that Cbfa1 is upregulated in the second branchial arches of Hoxa-2 mutant embryos suggesting that prevention of Cbfa1 induction might mediate Hoxa-2 inhibition of dermal bone formation during normal second arch development. The implications of these results on the patterning of the branchial area are discussed.


Subject(s)
Bone Development/physiology , Branchial Region/growth & development , Gene Expression Regulation, Developmental/genetics , Head/growth & development , Homeodomain Proteins/genetics , Neoplasm Proteins , Animals , Branchial Region/embryology , Cartilage/embryology , Cartilage/growth & development , Core Binding Factor Alpha 1 Subunit , DNA Probes , Embryo, Mammalian/cytology , Embryonic and Fetal Development , Genes, Homeobox/genetics , Head/embryology , High Mobility Group Proteins/genetics , Humans , In Situ Hybridization , Mice , Mice, Knockout , SOX9 Transcription Factor , Transcription Factors/genetics
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