Xavier Caubit

Chaetognath phylogenomics: a protostome with deuterostome-like development

Traditional textbook phylogeny splits bilaterians into protostomes and deuterostomes according to whether their mouth derives from the blastopore or not. This scheme has been largely confirmed by small subunit ribosomal RNA (SSU) molecular phylogeny. However, some phyla, such as the lophphorate phyla Phoronida and Brachiopoda as well as the Chaetognatha exhibit classical deuterostome embryological features such as formation of the mesoderm from the gut (enterocoely) and secondary opening of the mouth.

Three putative murine Teashirt orthologues specify trunk structures in Drosophila in the same way as the Drosophila teashirt gene.

Drosophila teashirt (tsh) functions as a region-specific homeotic gene that specifies trunk identity during embryogenesis. Based on sequence homology, three tsh-like (Tsh) genes have been identified in the mouse. Their expression patterns in specific regions of the trunk, limbs and gut raise the possibility that they may play similar roles to tsh in flies. By expressing the putative mouse Tsh genes in flies, we provide evidence that they behave in a very similar way to the fly tsh gene. First, ectopic expression of any of the three mouse Tsh genes, like that of tsh, induces head to trunk homeotic transformation. Second, mouse Tsh proteins can rescue both the homeotic and the segment polarity phenotypes of a tsh null mutant. Third, following ectopic expression, the three mouse Tsh genes affect the expression of the same target genes as tsh in the Drosophila embryo. Fourth, mouse Tsh genes, like tsh, are able to induce ectopic eyes in adult flies. Finally, all Tsh proteins contain a motif that recruits the C-terminal binding protein and contributes to their repression function. As no other vertebrate or fly protein has been shown to induce such effects upon ectopic expression, these results are consistent with the idea that the three mouse Tsh genes are functionally equivalent to the Drosophila tsh gene when expressed in developing Drosophila embryos.

Teashirt 3 Regulates Development of Neurons Involved in Both Respiratory Rhythm and Airflow Control

Neonatal breathing in mammals involves multiple neuronal circuits, but its genetic basis remains unclear. Mice deficient for the zinc finger protein Teashirt 3 (TSHZ3) fail to breathe and die at birth. Tshz3 is expressed in multiple areas of the brainstem involved in respiration, including the pre-Bötzinger complex (preBötC), the embryonic parafacial respiratory group (e-pF), and cranial motoneurons that control the upper airways. Tshz3 inactivation led to pronounced cell death of motoneurons in the nucleus ambiguus and induced strong alterations of rhythmogenesis in the e-pF oscillator. In contrast, the preBötC oscillator appeared to be unaffected. These deficits result in impaired upper airway function, abnormal central respiratory rhythm generation, and altered responses to pH changes. Thus, a single gene, Tshz3, controls the development of diverse components of the circuitry required for breathing.

Expression patterns of the three Teashirt-related genes define specific boundaries in the developing and postnatal mouse forebrain

We compare the expression patterns of the three mouse Teashirt (mTsh) genes during development of the forebrain and at a postnatal stage. During development, mTsh genes are expressed in domains that are restricted both dorsoventrally and rostrocaudally, with major changes in expression level coinciding with compartment boundaries. Striking complementarities in the distribution of mTsh transcripts were observed in the developing diencephalon, telencephalon, and olfactory bulb (OB). A mTsh1‐positive cell population is part of the DLX‐positive population localized in the dorsalmost portion of the lateral ganglionic eminence (dLGE). Comparison of the mTsh1 expression domain with the domains of Er81 and Islet1, which mark two distinct progenitor populations in the subventricular zone of the LGE, suggests that mTsh1 marks OB interneuron progenitors. Furthermore, the distinct expression patterns of mTsh1 and mTsh2 in the ventral LGE and the dLGE highlight the differential contributions of these structures to the striatum and the amydaloid complex. For Sey/Sey mutants, we show that Pax6 function is critical for the correct specification of the mTsh1+ population in the dLGE during embryogenesis. At postnatal stages in the OB, mTsh1 is expressed in granule and periglomerular cells, which originate from the subpallium during development. Furthermore, mTsh1+ cells line the walls of the anterior lateral ventricle, a region that gives rise to the interneurons that migrate in the rostral migratory streams and populate the OB postnatally. Our results suggest a role for mTsh genes in the establishment of regional identity and specification of cell types in the developing and adult forebrain. J. Comp. Neurol. 486:76–88, 2005.

Teashirt-3, a Novel Regulator of Muscle Differentiation, Associates with BRG1-associated Factor 57 (BAF57) to Inhibit Myogenin Gene Expression

In adult muscles and under normal physiological conditions, satellite cells are found in a quiescent state but can be induced to enter the cell cycle by signals resulting from exercise, injury-induced muscle regeneration, or specific disease states. Once activated, satellite cells proliferate, self-renew, and differentiate to form myofibers. In the present study, we found that the zinc finger-containing factor Teashirt-3 (TSHZ3) was expressed in quiescent satellite cells of adult mouse skeletal muscles. We showed that following treatment with cardiotoxin TSHZ3 was strongly expressed in satellite cells of regenerating muscles. Moreover, immunohistochemical analysis indicated that TSHZ3 was expressed in both quiescent and activated satellite cells on intact myofibers in culture. TSHZ3 expression was maintained in myoblasts but disappeared with myotube formation. In C2C12 myoblasts, we showed that overexpression of Tshz3 impaired myogenic differentiation and promoted the down-regulation of myogenin (Myog) and up-regulation of paired-box factor 7 (Pax7). Moreover, knockdown experiments revealed a selective effect of Tshz3 on Myog regulation, and transcriptional reporter experiments indicated that TSHZ3 repressed Myog promoter. We identified the BRG1-associated factor 57 (BAF57), a subunit of the SWI/SNF complex, as a partner of TSHZ3. We showed that TSHZ3 cooperated with BAF57 to repress MYOD-dependent Myog expression. These results suggest a novel mechanism for transcriptional repression by TSHZ3 in which TSHZ3 and BAF57 cooperate to modulate MyoD activity on the Myog promoter to regulate skeletal muscle differentiation.

Analysis of TSHZ2 and TSHZ3 genes in congenital pelvi-ureteric junction obstruction

BACKGROUND Congenital pelvi-ureteric junction obstruction (PUJO) affects 0.3% of human births. It may result from aberrant smooth muscle development in the renal pelvis, resulting in hydronephrosis. Mice that are null mutant for the Teashirt3 (Tshz3) gene exhibit congenital PUJO with defective smooth muscle differentiation and absent peristalsis in the proximal ureter. METHODS Given the phenotype of Tshz3 mutant mice, we considered that Teashirt genes, which code for a family of transcription factors, might represent candidate genes for human PUJO. To evaluate this possibility, we used in situ hydridization to analyse the three mammalian Tshz genes in mouse embryonic ureters and determined whether TSHZ3 was expressed in the human embryonic ureter. TSHZ2 and TSHZ3 were sequenced in index cases with non-syndromic PUJO. RESULTS Tshz2 and Tshz3 genes were detected in mouse ureters and TSHZ3 was expressed in the human embryonic renal pelvis. Direct sequencing of TSHZ2 and TSHZ3 did not identify any mutations in an initial cohort of 48 PUJO index cases, excluding these genes as a major cause of this condition. A polymorphic missense change (E469G) in TSHZ3 was identified at a residue highly conserved throughout evolution in all Teashirt proteins, although subsequently no significant difference between the E469G allele frequency in Albanian and Macedonian PUJO index cases (3.2%) versus 633 control individuals (1.7%) was found (P = 0.18). CONCLUSIONS Mutations in TSHZ2 and TSHZ3 are not a major cause of PUJO, at least in Albanian and Macedonian populations. Expression of these genes in the human fetal ureter emphasizes the importance of analysing these genes in other groups of patients with renal tract malformations.

TSHZ3 deletion causes an autism syndrome and defects in cortical projection neurons

TSHZ3, which encodes a zinc-finger transcription factor, was recently positioned as a hub gene in a module of the genes with the highest expression in the developing human neocortex, but its functions remained unknown. Here we identify TSHZ3 as the critical region for a syndrome associated with heterozygous deletions at 19q12-q13.11, which includes autism spectrum disorder (ASD). In Tshz3-null mice, differentially expressed genes include layer-specific markers of cerebral cortical projection neurons (CPNs), and the human orthologs of these genes are strongly associated with ASD. Furthermore, mice heterozygous for Tshz3 show functional changes at synapses established by CPNs and exhibit core ASD-like behavioral abnormalities. These findings highlight essential roles for Tshz3 in CPN development and function, whose alterations can account for ASD in the newly defined TSHZ3 deletion syndrome.