Identification of critical regions for clinical features of distal 10q deletion syndrome.

Array comparative genomic hybridization studies were performed to further characterize cytogenetic abnormalities found originally by karyotype and fluorescence in situ hybridization in five clinical cases of distal 10q deletions, including several with complex cytogenetic rearrangements and one with a partial male-to-female sex-reversal phenotype. These results have enabled us to narrow the previously proposed critical regions for the craniofacial, urogenital, and neuropsychiatric disease-related manifestations associated with distal 10q deletion syndrome. Furthermore, we propose that haploinsufficiency of the DOCK1 gene may play a crucial role in the pathogenesis of the 10q deletion syndrome. We hypothesize that alteration of DOCK1 and/or other genes involved in regulation and signaling of multiple pathways can explain the wide range of phenotypic variability between patients with similar or identical cytogenetic abnormalities.

Acute extrapyramidal syndrome in mild ornithine transcarbamylase deficiency: metabolic stroke involving the caudate and putamen without metabolic decompensation.

UNLABELLED: A 6-year-old male with partial ornithine transcarbamylase (OTC) deficiency had acute and rapidly progressive symmetrical swelling of the head of the caudate nuclei and putamina. Clinical presentation was ataxia and dysarthria progressing to seizures and coma; these symptoms gradually resolved with supportive management. Although he had been recently treated for mild hyperammonemia, there was no evidence of acute metabolic decompensation prior to presentation, and plasma ammonia and amino acids were consistent with good metabolic control. This case is novel in that the neurological insult affected the neostriatum of the basal ganglia and the episode occurred in the absence of an apparent metabolic abnormality, unique observations in a patient with OTC deficiency. CONCLUSION: This case suggests that the pathophysiology of metabolic stroke is complicated. It also argues for an evaluation for metabolic stroke in patients with known inborn errors of metabolism who present with unusual neurological symptoms in the absence of biochemical abnormalities. Similarly, this case suggests that patients presenting with unexplained neurological insults might benefit from an evaluation for an inborn error of metabolism.

SF-1, DAX-1, and acd: molecular determinants of adrenocortical growth and steroidogenesis.

The formation of the adrenal cortex in humans is notable for the presence of two discrete zones, the fetal zone (FZ) which regresses soon after birth and the definitive zone (DZ) which gives rise to the classic steroidogenic zones of the adult cortex. Mice possess an analogous structure to the FZ referred to as the X-zone (XZ) which regresses at puberty in the male and during the first pregnancy in the female. Similar to the human FZ in X-linked Congenital Adrenal Hypoplasia caused by loss of function mutations in DAX-1 (Dosage-sensitive sex reversal-Adrenal hypoplasia congenita critical region on the X chromosome), the mouse XZ does not regress when DAX-1 is mutated. Only in humans with DAX-1 mutations, however, is the DZ small and hypofunctional. Patients and mice with SF-1 mutations have complete adrenal aplasia with absence of both the DZ and FZ/XZ. Lastly, the phenotype of the Autosomal Recessive Adrenocortical Dysplasia (acd) mouse is strikingly similar to human Miniature Adult Congenital Adrenal Hypoplasia, lacking an XZ/FZ and possessing a dysfunctional DZ. Current work has addressed the regulation of SF-1 and DAX-1 dependent adrenocortical growth and steroidogenesis in vivo utilizing mouse models of simple and combined SF-1 and DAX-1 deficiency. In addition, the model of compensatory adrenal growth in SF-1 haplo-insufficient mice has been applied to evaluate the potential role of SF-1 in adrenocortical proliferation. Additional efforts aim to positionally clone the acd gene, predicated on the hypothesis that it is a critical component of the adrenal developmental cascade.

Townes-Brocks syndrome versus expanded spectrum hemifacial microsomia: review of eight patients and further evidence of a "hot spot" for mutation in the SALL1 gene.

PURPOSE: It can be difficult to differentiate clinically between hemifacial microsomia (HFM) and Townes-Brocks syndrome (TBS). The distinction is important because TBS is inherited as an autosomal dominant trait, whereas HFM is sporadic. METHODS: We performed a retrospective analysis of eight patients with HFM-expanded spectrum and anal anomalies to determine whether this subset has TBS. RESULTS: Two patients had major phenotypic findings of TBS. Sequencing of SALL1, the gene mutated in TBS, in four of the eight patients revealed one with a C --> T transition (resulting in a nonsense mutation R276X) at a previously identified mutational "hot spot." CONCLUSION: Patients with overlapping features of both syndromes should be screened for SALL1 mutations.

Implementing transgenic and embryonic stem cell technology to study gene expression, cell-cell interactions and gene function.

This review highlights the use of transgenic mice and gene targeting in the study of reproduction, pituitary gene expression, and cell lineage. Since 1980 numerous applications of transgenic animal technology have been reported. Altered phenotypes resulting from transgene expression demonstrated that introduced genes can exert profound effects on animal physiology. Transgenic mice have been important for the study of hormonal and developmental control of gene expression because gene expression in whole animals often requires more DNA sequence information than is necessary for expression in cell cultures. This point is illustrated by studies of pituitary glycoprotein hormone alpha- and beta-subunit gene expression (Kendall et al., Mol Endocrinol 1994; in press [1]. Transgenic mice have also been invaluable for producing animal models of cancer and other diseases and testing the efficacy of gene therapy. In addition, cell-cell interactions and cell lineage relationships have been explored by cell-specific expression of toxin genes in transgenic mice. Recent studies suggest that attenuated and inducible toxins hold promise for future transgene ablation experiments. Since 1987, embryonic stem (ES) cell technology has been used to create numerous mouse strains with targeted gene alterations, contributing enormously to our understanding of the functional importance of individual genes. For example, the unexpected development of gonadal tumors in mice with a targeted disruption of the inhibin gene revealed a potential role for inhibin as a tumor suppressor (Matzuk et al., Nature 1992:360: 313-319 [2]. The transgenic and ES cell technologies will undoubtedly continue to expand our understanding and challenge our paradigms in reproductive biology.

Expression of corticotropin-releasing hormone transgenes in neurons of adult and developing mice.

The DNA sequences important for cell-specific expression and developmental regulation of corticotropin-releasing hormone (CRH) were analyzed in transgenic mice. A construct containing 0.5 kb of CRH 5' flanking DNA linked to the chloramphenicol acetyltransferase reporter gene was expressed in many brain regions and in several ectopic peripheral sites, suggesting that this portion of the CRH gene contains basal promoter activity but lacks DNA elements necessary for appropriate tissue specificity. Cell specificity of transgene expression was examined with a CRH-beta-galactosidase reporter construct containing the same 0.5-kb CRH promoter fragment, but also including the CRH structural gene and 2 kb of CRH 3' flanking DNA. Transgene expression was observed in inappropriate regions of the brain, but no expression was detected in peripheral tissues, suggesting that these additional CRH sequences suppress inappropriately high levels of peripheral expression. Cell-specific expression improved significantly with the inclusion of 8.7 kb of CRH 5' flanking DNA. Individual transgenic lines exhibited expression in a number of the major CRH neuronal groups including the paraventricular nucleus, medial geniculate nucleus, inferior olivary nucleus, and Barrington's nucleus. Transgene expression was properly activated in Barrington's nucleus during development. This study demonstrates that the regulatory control of cell-specific and developmentally appropriate CRH expression is complex, utilizing multiple DNA sequence elements located upstream and downstream of the CRH transcription start site.

Differential expression of corticotropin-releasing hormone in developing mouse embryos and adult brain.

CRH mRNA was detected by in situ hybridization histochemistry in numerous regions of the adult mouse brain, including most prominently the paraventricular nucleus (PVN) of the hypothalamus, the inferior olivary nucleus, and Barrington's nucleus. After adrenalectomy, steady state CRH mRNA levels increased 1.7-fold, specifically in the PVN, consistent with reports of negative glucocorticoid regulation of CRH expression in the rat PVN. Ontogenetic analysis of CRH expression in fetal and neonatal mouse brain demonstrated CRH mRNA in PVN, Barrington's nucleus, olivary complex, and amygdaloid primordia on embryonic day 13.5. In contrast, CRH mRNA was not detectable in the cortex until after birth. CRH expression also exhibited differential regulation in ontogeny. CRH mRNA reached adult levels at markedly different times of development in each brain region, and CRH expression was reduced specifically in the PVN just before birth and the stress hyporesponsive period. High levels of CRH mRNA were present transiently in the developing lung and celiac ganglion. The novel findings of CRH expression in fetal lung during the period of glucocorticoid-induced lung maturation and in celiac ganglion during development of the sympathetic nervous system indicate that CRH may have some important developmental functions in addition to its role in activation of the stress response.