Two vertebrate homeobox genes related to the Drosophila empty spiracles gene are expressed in the embryonic cerebral cortex.

We cloned two homeobox genes, Emx1 and Emx2, related to empty spiracles, a gene expressed in very anterior body regions during early Drosophila embryogenesis, and studied their expression in mouse embryos. Emx1 expression is detectable from day 9.5 of gestation whereas Emx2 appears to be already expressed in 8.5 day embryos. Both genes are expressed in the presumptive cerebral cortex and olfactory bulbs. Emx1 is expressed exclusively there, whereas Emx2 is also expressed in some neuroectodermal areas in embryonic head including olfactory placodes in earlier stages and olfactory epithelia later in development.     

Orthodenticle and empty spiracles genes are expressed in a segmental pattern in chelicerates

Members of the orthodenticle (otd/Otx) and empty spiracles (ems/Emx) gene families are head gap genes that encode homeodomain-containing DNA-binding proteins. Although numerous studies show their central role in developmental processes in brain specification, a surprisingly high number of other developmental processes have been shown to involve their expression. In this paper, we report the identification and expression of ems and otd in two chelicerate species: a scorpion, Euscorpius flavicaudis (Chactidae, Scorpiona, Arachnida, Euchelicerata) and a spider, Tegenaria saeva (Aranea, Arachnida, Euchelicerata). We show that both ems and otd are expressed not only in an anterior head domain but also along the entire anterior–posterior axis during embryonic development. The expression patterns for both genes are typically segmental and concern neurectodermal territories. During patterning of the opisthosoma, ems and otd are expressed in the lateral ectoderm just anterior to the limb bud primordia giving rise to respiratory organs and spinnerets (spider). This common pattern found in two divergent species thus appears to be a conserved character of chelicerates. These results are discussed in terms of evolutionary origin of respiratory organs and/or functional pathway recruitment.     

Structural and functional diversity of homeodomain genes of the orthodenticle and empty spiracles classes in Craniata

Despite extensively divergent morphologies, the patterning of the embryonic brain is controlled by highly conserved genetic networks. Orthodenticle and empty spiracles-related homeodomain genes, which are expressed in insects as in vertebrates in anteriormost neuromeres of the embryonic brain, provide examples of such conservations. In gnathostomes, they form small multigene families, each containing three well-characterised orthology classes. In mice, paralogous genes play very different roles in the development of cephalic regions. Some of these roles are probably ancient and conserved in all chordates, while others, like the morphogenesis of gnathostomespecific characters, correspond to much more diversified functions. Genetic analyses in mice together with comparative analyses in a broad range of vertebrates provide new possibilities to investigate the molecular mechanisms which underlie these functional diversifications.     

Expression, regulation and function of the homeobox gene empty spiracles in brain and ventral nerve cord development of Drosophila

We analyse the role of the empty spiracles (ems) gene in embryonic brain and ventral nerve cord development. ems is differentially expressed in the neurectoderm of the anterior head versus the trunk region of early embryos. A distal enhancer region drives expression in the deutocerebral brain anlage and a proximal enhancer region drives expression in the VNC and tritocerebral brain anlage. Mutant analysis indicates that in the anterior brain ems is necessary for regionalized neurogenesis in the deutocerebral and tritocerebral anlagen. In the posterior brain and VNC ems is necessary for correct axonal pathfinding of specific interneurons. Rescue experiments indicate that the murine Emx2 gene can partially replace the fly ems gene in CNS development.     

Regulation of CLV3 expression by two homeobox genes in Arabidopsis

The ability of meristems to continuously produce new organs depends on the activity of their stem cell populations, which are located at the meristem tip. In Arabidopsis, the size of the stem cell domain is regulated by two antagonistic activities. The WUS (WUSCHEL) gene, encoding a homeodomain protein, promotes the formation and maintenance of stem cells. These stem cells express CLV3 (CLAVATA3), and signaling of CLV3 through the CLV1/CLV2 receptor complex restricts WUS activity. Homeostasis of the stem cell population may be achieved through feedback regulation, whereby changes in stem cell number result in corresponding changes in CLV3 expression levels, and adjustment of WUS expression via the CLV signal transduction pathway. We have analyzed whether expression of CLV3 is controlled by the activity of WUS or another homeobox gene, STM (SHOOT MERISTEMLESS), which is required for stem cell maintenance. We found that expression of CLV3 depends on WUS function only in the embryonic shoot meristem. At later developmental stages, WUS promotes the level of CLV3 expression, together with STM. Within a meristem, competence to respond to WUS activity by expressing CLV3 is restricted to the meristem apex.     

Craniofacial disorders caused by mutations in homeobox genes MSX1 and MSX2

The molecular biology of the homeobox genes MSX1 and MSX2 is reviewed. In a selective type of tooth agenesis, an MSX1 G --> C transversion results in a missense mutation Arg31Pro. The phenotype is due to haploinsufficiency. Boston-type craniosynostosis involves an MSX2 C --> A transversion, resulting in a missense mutation Pro7His. Three different mutations on MSX2 cause parietal foramina by haploinsufficiency. These mutations, which result in decreased parietal ossification, are in marked contrast to the gain-of-function mutation for Boston-type craniosynostosis, which results in increased sutural ossification.     

The Cdx-1 and Cdx-2 homeobox genes in the intestine.

The past years have witnessed an increasing number of reports relative to homeobox genes in endoderm-derived tissues. In this review, we focus on the caudal-related Cdx-1 and Cdx-2 homeobox genes to give an overview of the in vivo, in vitro, and ex vivo approaches that emphasize their primary role in intestinal development and in the control of intestinal cell proliferation, differentiation, and identity. The participation of these genes in colon tumorigenesis and their identification as important actors of the oncogenic process are also discussed.     

Expression and function of the empty spiracles gene in olfactory sense organ development of Drosophila melanogaster

In Drosophila, the cephalic gap gene empty spiracles plays key roles in embryonic patterning of the peripheral and central nervous system. During postembryonic development, it is involved in the development of central olfactory circuitry in the antennal lobe of the adult. However, its possible role in the postembryonic development of peripheral olfactory sense organs has not been investigated. Here, we show that empty spiracles acts in a subset of precursors that generate the olfactory sense organs of the adult antenna. All empty spiracles-expressing precursor cells co-express the proneural gene amos and the early patterning gene lozenge. Moreover, the expression of empty spiracles in these precursor cells is dependent on both amos and lozenge. Functional analysis reveals two distinct roles of empty spiracles in the development of olfactory sense organs. Genetic interaction studies in a lozenge-sensitized background uncover a requirement of empty spiracles in the formation of trichoid and basiconic olfactory sensilla. MARCM-based clonal mutant analysis reveals an additional role during axonal targeting of olfactory sensory neurons to glomeruli within the antennal lobe. Our findings on empty spiracles action in olfactory sense organ development complement previous studies that demonstrate its requirement in olfactory interneurons and, taken together with studies on the murine homologs of empty spiracles, suggest that conserved molecular genetic programs might be responsible for the formation of both peripheral and central olfactory circuitry in insects and mammals.     

Three knotted1-like homeobox genes in Arabidopsis

Five arabidopsis kn1-like homeobox genes were cloned through low-stringency screening of Arabidopsis cDNA libraries with the kn1 homeobox from maize. These five genes were named KNAT1-5 (for kn1-like Arabidopsis thaliana). An analysis of KNAT1 and 2 has been presented previously [19]. Here we present an analysis of the genes KNAT3, 4 and 5. On the basis of sequence and expression patterns, these three genes belong to the class II subfamily of kn1-like homeobox genes [16]. Low-stringency Southern analysis suggests several additional members of the class II genes exist in the Arabidopsis genome. The predicted amino acid sequences of the three genes share extensive homology outside of the homeodomain, including 84% between KNAT3 and KNAT4. Northern analysis shows that although all three genes are expressed in all tissues examined, the level of KNAT3 RNA is highest in young siliques, inflorescences and roots, KNAT4 RNA level is strongest in leaves and young siliques, and KNAT5 RNA level is highest in roots. The specificity of these patterns was confirmed by RNA fingerprint analysis. KNAT3 and 4 are light-regulated as they show reduced expression in etiolated seedlings and also in hy3, cop1 and det1 mutant backgrounds.     

Formation and specification of distal leg segments in Drosophila by dual Bar homeobox genes, BarH1 and BarH2

Here, we show that BarH1 and BarH2, a pair of Bar homeobox genes, play essential roles in the formation and specification of the distal leg segments of Drosophila. In early third instar, juxtaposition of Bar-positive and Bar-negative tissues causes central folding that may separate future tarsal segments 2 from 3, while juxtaposition of tissues differentially expressing Bar homeobox genes at later stages gives rise to segmental boundaries of distal tarsi including the tarsus/pretarsus boundary. Tarsus/pretarsus boundary formation requires at least two different Bar functions, early antagonistic interactions with a pretarsus-specific homeobox gene, aristaless, and the subsequent induction of Fas II expression in pretarsus cells abutting tarsal segment 5. Bar homeobox genes are also required for specification of distal tarsi. Bar expression requires Distal-less but not dachshund, while early circular dachshund expression is delimited interiorly by BarH1 and BarH2.     

Regulation of retinal cone bipolar cell differentiation and photopic vision by the CVC homeobox gene Vsx1

Cone bipolar cells of the vertebrate retina connect photoreceptors with ganglion cells to mediate photopic vision. Despite this important role, the mechanisms that regulate cone bipolar cell differentiation are poorly understood. VSX1 is a CVC domain homeoprotein specifically expressed in cone bipolar cells. To determine the function of VSX1, we generated Vsx1 mutant mice and found that Vsx1 mutant retinal cells form but do not differentiate a mature cone bipolar cell phenotype. Electrophysiological studies demonstrated that Vsx1 mutant mice have defects in their cone visual pathway, whereas the rod visual pathway was unaffected. Thus, Vsx1 is required for cone bipolar cell differentiation and regulates photopic vision perception.