Children's Pronoun Interpretation Problems Are Related to Theory of Mind and Inhibition, But Not Working Memory.

In several languages, including English and Dutch, children's acquisition of the interpretation of object pronouns (e.g., him) is delayed compared to that of reflexives (e.g., himself). Various syntactic and pragmatic explanations have been proposed to account for this delay in children's acquisition of pronoun interpretation. This study aims to provide more insight into this delay by investigating potential cognitive mechanisms underlying this delay. Dutch-speaking children between 6 and 12 years old with autism spectrum disorder (ASD; n = 47), attention-deficit/hyperactivity disorder (ADHD; n = 36) or typical development (TD; n = 38) were tested on their interpretation and production of object pronouns and reflexives and on theory of mind, working memory, and response inhibition. It was found that all three groups of children had difficulty with pronoun interpretation and that their performance on pronoun interpretation was associated with theory of mind and inhibition. These findings support an explanation of object pronoun interpretation in terms of perspective taking, according to which listeners need to consider the speaker's perspective in order to block coreference between the object pronoun and the subject of the same sentence. Unlike what is predicted by alternative theoretical accounts, performance on pronoun interpretation was not associated with working memory, and the children made virtually no errors in their production of object pronouns. As the difficulties with pronoun interpretation were similar for children with ASD, children with ADHD and typically developing children, this suggests that certain types of perspective taking are unaffected in children with ASD and ADHD.

Complex Inferential Processes Are Needed for Implicature Comprehension, but Not for Implicature Production.

Upon hearing "Some of Michelangelo's sculptures are in Rome," adults can easily generate a scalar implicature and infer that the intended meaning of the utterance corresponds to "Some but not all Michelangelo's sculptures are in Rome." Comprehension experiments show that preschoolers struggle with this kind of inference until at least 5 years of age. Surprisingly, the few studies having investigated children's production of scalar expressions like some and all suggest that production is adult-like already in their third year of life. Thus, children's production of implicatures seems to develop at least 2 years before their comprehension of implicatures. In this paper, we present a novel account of scalar implicature generation in the framework of Bidirectional Optimality Theory: the Asymmetry Account. We show that the production-comprehension asymmetry is predicted to emerge because the comprehension of some requires the hearer to consider the speaker's perspective, but the production of some does not require the speaker to consider the hearer's perspective. Hence, children's comprehension of scalar expressions, but not their production of scalar expressions, is predicted to be related to their theory of mind development. Not possessing fully developed theory of mind abilities yet, children thus have difficulty in comprehending scalar expressions such as some in an adult-like way. Our account also explains why variable performance is found in experimental studies testing children's ability to generate scalar implicatures; moreover, it describes the differences between children's and adults' implicature generation in terms of their ability to recursively apply theory of mind; finally, it sheds new light on the question why the interpretation of numerals does not require implicature generation.

Narrative production in children with autism spectrum disorder (ASD) and children with attention-deficit/hyperactivity disorder (ADHD): Similarities and differences.

The present study focuses on the similarities and differences in language production between children with autism spectrum disorder (ASD) and children with attention-deficit/hyperactivity disorder (ADHD). In addition, we investigated whether Theory of Mind (ToM), working memory, and response inhibition are associated with language production. Narratives, produced by 106 Dutch-speaking children (36 with ASD, 34 with ADHD, and 36 typically developing) aged 6 to 12 during ADOS assessment, were examined on several linguistic measures: verbal productivity, speech fluency, syntactic complexity, lexical semantics, and discourse pragmatics. Children were tested on ToM, working memory, and response inhibition and parents filled in the Children's Communication Checklist (CCC-2). Gold-standard diagnostic measures (Autism Diagnostic Observation Schema [ADOS], Autism Diagnostic Interview Revised [ADI-R], and the Parent Interview for Child Symptoms [PICS]) were administered to all children to confirm diagnosis. Regarding similarities, both clinical groups showed impairments in narrative performance relative to typically developing children. These were confirmed by the CCC-2. These impairments were not only present on pragmatic measures, such as the inability to produce a narrative in a coherent and cohesive way, but also on syntactic complexity and their production of repetitions. As for differences, children with ADHD but not children with ASD showed problems in their choice of referring expressions and speech fluency. ToM and working memory performance but not response inhibition were associated with many narrative skills, suggesting that these cognitive mechanisms explain some of the impairments in language production. We conclude that children with ASD and children with ADHD manifest multiple and diverse language production problems, which may partly relate to their problems in ToM and working memory. (PsycINFO Database Record

Who Is He? Children with ASD and ADHD Take the Listener into Account in Their Production of Ambiguous Pronouns.

During conversation, speakers constantly make choices about how specific they wish to be in their use of referring expressions. In the present study we investigate whether speakers take the listener into account or whether they base their referential choices solely on their own representation of the discourse. We do this by examining the cognitive mechanisms that underlie the choice of referring expression at different discourse moments. Furthermore, we provide insights into how children with Autism Spectrum Disorder (ASD) and Attention Deficit Hyperactivity Disorder (ADHD) use referring expressions and whether their use differs from that of typically developing (TD) children. Children between 6 and 12 years old (ASD: n=46; ADHD: n=37; TD: n=38) were tested on their production of referring expressions and on Theory of Mind, response inhibition and working memory. We found support for the view that speakers take the listener into account when choosing a referring expression: Theory of Mind was related to referential choice only at those moments when speakers could not solely base their choice on their own discourse representation to be understood. Working memory appeared to be involved in keeping track of the different referents in the discourse. Furthermore, we found that TD children as well as children with ASD and children with ADHD took the listener into account in their choice of referring expression. In addition, children with ADHD were less specific than TD children in contexts with more than one referent. The previously observed problems with referential choice in children with ASD may lie in difficulties in keeping track of longer and more complex discourses, rather than in problems with taking into account the listener.

Combined MYC and P53 defects emerge at medulloblastoma relapse and define rapidly progressive, therapeutically targetable disease.

We undertook a comprehensive clinical and biological investigation of serial medulloblastoma biopsies obtained at diagnosis and relapse. Combined MYC family amplifications and P53 pathway defects commonly emerged at relapse, and all patients in this group died of rapidly progressive disease postrelapse. To study this interaction, we investigated a transgenic model of MYCN-driven medulloblastoma and found spontaneous development of Trp53 inactivating mutations. Abrogation of p53 function in this model produced aggressive tumors that mimicked characteristics of relapsed human tumors with combined P53-MYC dysfunction. Restoration of p53 activity and genetic and therapeutic suppression of MYCN all reduced tumor growth and prolonged survival. Our findings identify P53-MYC interactions at medulloblastoma relapse as biomarkers of clinically aggressive disease that may be targeted therapeutically.

How to use the ADI-R for classifying autism spectrum disorders? Psychometric properties of criteria from the literature in 1,204 Dutch children.

The algorithm of the Autism Diagnostic Interview-Revised provides criteria for autism versus non-autism according to DSM-IV. Criteria for the broader autism spectrum disorders are needed. This study investigated the validity of seven sets of criteria from the literature, in 1,204 Dutch children (aged 3-18 years) with and without mental retardation. The original criteria (Rutter et al. in ADI-R Autism Diagnostic Interview Revised. Manual. Western Psychological Services, Los Angeles, 2003) well discriminated ASD from non-ASD in MR. All other criteria (IMGSAC in Am Soc Hum Genet 69:570-581 2001; Sung et al. in Am J Hum Genet 76: 68-81, 2005; Risi et al. in J Am Acad Child Adolesc Psychiatry 45: 1094-1103, 2006) were sensitive at the cost of specificity, bearing the risk of overinclusiveness. In the group without MR, clinicians should decide whether sensitivity or specificity is aimed for, to choose the appropriate criteria. Including the Autism Diagnostic Observation Schedule revised algorithms in the classification, the specificity increases, at the cost of sensitivity. This study adds to a more valid judgment on which criteria to use for specific objectives.

Regulation of angiogenesis by Eph-ephrin interactions.

The large families of Eph receptor tyrosine kinases and their ephrin ligands transduce signals in a cell-cell contact-dependent fashion and thereby coordinate the growth, differentiation, and patterning of almost every organ and tissue. Eph-ephrin interactions can trigger a wide array of cellular responses, including cell adhesion, boundary formation, and repulsion. The exact mechanisms leading to this diversity of responses are unclear but appear to involve differential signaling, proteolytic cleavage of ephrins, and endocytosis of the ligand-receptor complex. In the developing cardiovascular system, Eph and ephrin molecules control the angiogenic remodeling of blood vessels and lymphatic vessels and play essential roles in endothelial cells as well as in supporting pericytes and vascular smooth muscle cells. Recent evidence suggests that Ephs and ephrins may also be involved in pathological angiogenesis, in particular, the neovascularization of tumors. Consequently, the expression, interactions, or signaling of Eph-ephrin molecules might be targets for future therapeutic approaches.

Targeted mutation reveals essential functions of the homeodomain transcription factor Shox2 in sinoatrial and pacemaking development.

BACKGROUND: Identifying molecular pathways regulating the development of pacemaking and coordinated heartbeat is crucial for a comprehensive mechanistic understanding of arrhythmia-related diseases. Elucidation of these pathways has been complicated mainly by an insufficient definition of the developmental structures involved in these processes and the unavailability of animal models specifically targeting the relevant tissues. Here, we report on a highly restricted expression pattern of the homeodomain transcription factor Shox2 in the sinus venosus myocardium, including the sinoatrial nodal region and the venous valves. METHODS AND RESULTS: To investigate its function in vivo, we have generated mouse lines carrying a targeted mutation of the Shox2 gene. Although heterozygous animals did not exhibit obvious defects, homozygosity of the targeted allele led to embryonic lethality at 11.5 to 13.5 dpc. Shox2-/- embryos exhibited severe hypoplasia of the sinus venosus myocardium in the posterior heart field, including the sinoatrial nodal region and venous valves. We furthermore demonstrate aberrant expression of connexin 40 and connexin 43 and the transcription factor Nkx2.5 in vivo specifically within the sinoatrial nodal region and show that Shox2 deficiency interferes with pacemaking function in zebrafish embryos. CONCLUSIONS: From these results, we postulate a critical function of Shox2 in the recruitment of sinus venosus myocardium comprising the sinoatrial nodal region.

Function and regulation of Alx4 in limb development: complex genetic interactions with Gli3 and Shh.

The role of the aristaless-related homeobox gene Alx4 in antero-posterior (AP-) patterning of the developing vertebrate limb has remained somewhat elusive. Polydactyly of Alx4 mutant mice is known to be accompanied by ectopic anterior expression of genes like Shh, Fgf4 and 5'Hoxd. We reported previously that polydactyly in Alx4 mutant mice requires SHH signaling, but we now show that in early Alx4-/- limb buds the anterior ectopic expression of Fgf4 and Hoxd13, and therefore disruption of AP-patterning, occurs independently of SHH signaling. To better understand how Alx4 functions in the pathways that regulate AP-patterning, we also studied genomic regulatory sequences that are capable of directing expression of a reporter gene in a pattern corresponding to endogenous Alx4 expression in anterior limb bud mesenchyme. We observed, as expected for authentic Alx4 expression, expansion of reporter construct expression in a Shh-/- background. Total lack of reporter expression in a Gli3-/- background confirms the existence of Gli3-dependent and -independent Alx4 expression in the limb bud. Apparently, these two modules of Alx4 expression are linked to dissimilar functions.

Genetics of shoulder girdle formation: roles of Tbx15 and aristaless-like genes.

The diverse cellular contributions to the skeletal elements of the vertebrate shoulder and pelvic girdles during embryonic development complicate the study of their patterning. Research in avian embryos has recently clarified part of the embryological basis of shoulder formation. Although dermomyotomal cells provide the progenitors of the scapular blade, local signals appear to have an essential guiding role in this process. These signals differ from those that are known to pattern the more distal appendicular skeleton. We have studied the impact of Tbx15, Gli3, Alx4 and related genes on formation of the skeletal elements of the mouse shoulder and pelvic girdles. We observed severe reduction of the scapula in double and triple mutants of these genes. Analyses of a range of complex genotypes revealed aspects of their genetic relationship, as well as functions that had been previously masked due to functional redundancy. Tbx15 and Gli3 appear to have synergistic functions in formation of the scapular blade. Scapular truncation in triple mutants of Tbx15, Alx4 and Cart1 indicates essential functions for Alx4 and Cart1 in the anterior part of the scapula, as opposed to Gli3 function being linked to the posterior part. Especially in Alx4/Cart1 mutants, the expression of markers such as Pax1, Pax3 and Scleraxis is altered prior to stages when anatomical aberrations are visible in the shoulder region. This suggests a disorganization of the proximal limb bud and adjacent flank mesoderm, and is likely to reflect the disruption of a mechanism providing positional cues to guide progenitor cells to their destination in the pectoral girdle.

Dorsoventral patterning of the mouse coat by Tbx15.

Many members of the animal kingdom display coat or skin color differences along their dorsoventral axis. To determine the mechanisms that control regional differences in pigmentation, we have studied how a classical mouse mutation, droopy ear (de(H)), affects dorsoventral skin characteristics, especially those under control of the Agouti gene. Mice carrying the Agouti allele black-and-tan (a(t)) normally have a sharp boundary between dorsal black hair and yellow ventral hair; the de(H) mutation raises the pigmentation boundary, producing an apparent dorsal-to-ventral transformation. We identify a 216 kb deletion in de(H) that removes all but the first exon of the Tbx15 gene, whose embryonic expression in developing mesenchyme correlates with pigmentary and skeletal malformations observed in de(H)/de(H) animals. Construction of a targeted allele of Tbx15 confirmed that the de(H) phenotype was caused by Tbx15 loss of function. Early embryonic expression of Tbx15 in dorsal mesenchyme is complementary to Agouti expression in ventral mesenchyme; in the absence of Tbx15, expression of Agouti in both embryos and postnatal animals is displaced dorsally. Transplantation experiments demonstrate that positional identity of the skin with regard to dorsoventral pigmentation differences is acquired by E12.5, which is shortly after early embryonic expression of Tbx15. Fate-mapping studies show that the dorsoventral pigmentation boundary is not in register with a previously identified dermal cell lineage boundary, but rather with the limb dorsoventral boundary. Embryonic expression of Tbx15 in dorsolateral mesenchyme provides an instructional cue required to establish the future positional identity of dorsal dermis. These findings represent a novel role for T-box gene action in embryonic development, identify a previously unappreciated aspect of dorsoventral patterning that is widely represented in furred mammals, and provide insight into the mechanisms that underlie region-specific differences in body morphology.

Progression of vertebrate limb development through SHH-mediated counteraction of GLI3.

Distal limb development and specification of digit identities in tetrapods are under the control of a mesenchymal organizer called the polarizing region. Sonic Hedgehog (SHH) is the morphogenetic signal produced by the polarizing region in the posterior limb bud. Ectopic anterior SHH signaling induces digit duplications and has been suspected as a major cause underlying congenital malformations that result in digit polydactyly. Here, we report that the polydactyly of Gli3-deficient mice arises independently of SHH signaling. Disruption of one or both Gli3 alleles in mouse embryos lacking Shh progressively restores limb distal development and digit formation. Our genetic analysis indicates that SHH signaling counteracts GLI3-mediated repression of key regulator genes, cell survival, and distal progression of limb bud development.