How clinicians add to knowledge of development.

Studies of human birth defects and developmental disorders have made major contributions to our understanding of development. Rare human syndromes have allowed identification of important developmental genes, and revealed mechanisms such as uniparental disomy and unstable trinucleotide repeats that were not suspected from animal studies. Some aspects of development, in particular cognitive development, can only be studied in human beings. Basic developmental mechanisms are very highly conserved across a very wide range of animals, making for a rich interplay between animal and human studies. Often, clinical studies identify a gene, or suggest a hypothesis, that can then be investigated in animals.

Diffuse plaques in the striatum in Alzheimer disease (AD): relationship to the striatal mosaic and selected neuropeptide markers.

Although neuritic and diffuse plaques generally coexist in Alzheimer disease (AD) neocortex, the predominance of diffuse plaques in the striatum prompted us to explore the potential influence of striatal features such as the striatal mosaic on beta-amyloid (A beta) deposition. Using double immunohistochemistry with an antibody to A beta in combination with antibodies to met-enkephalin or somatostatin, we investigated the relationship between diffuse plaques and neuropeptide distribution in the dorsal striatum. This relationship was examined in "pure" AD cases as well as in AD cases with coexisting Parkinson disease (PD) pathology, i.e. nigral degeneration and Lewy bodies at any site (AD + PD). Despite the presence of numerous A beta-positive diffuse plaques in both groups, the mosaic pattern, as exemplified by met-enkephalin-immunoreactive patches, was preserved. No obvious association was observed between the plaques and met-enkephalin-positive patches or somatostatin-immunoreactive neurons. Tyrosine hydroxylase immunoreactivity in the matrix was, however, diminished in AD + PD, most likely reflecting the nigral degeneration in these cases. Overall, these observations suggest that neostriatal A beta deposition in AD is not influenced by environmental factors associated with the striatal mosaic.

Cell mosaic patterns in the native and regenerated inner retina of zebrafish: implications for retinal assembly.

In part because of its laminar organization and morphologically distinct cell populations, the vertebrate retina has often been used as a system for investigating the assembly of neural structures. The retinas of adult teleost fish, because they grow throughout life and can regenerate following an injury, provide an especially attractive model system for such investigations. In an effort to provide a quantitative foundation for testing hypotheses regarding the mechanisms of pattern formation during growth and regeneration of the vertebrate retina, nearest neighbor and auto-correlation analyses were used to examine the mosaic patterns of eight inner retinal cell groups in the native and regenerated retina of adult zebrafish. In both native and regenerated retina, the mosaic patterns of most inner retinal cells are non-random. However, regenerated mosaics tend toward significantly lower nearest neighbor distances, less orderly patterns, and more variable radial locations than their native retina counterparts. The individual cell groups in both native and regenerated inner retina are likely to be spatially distributed independently. The results support the hypotheses that, in the adult zebrafish: 1) distinct inner retinal cell groups of native retina are also present in regenerated retina; 2) the assembly of inner retinal cell mosaics is controlled by non-random spatial organizing mechanisms during development, growth, and regeneration; and 3) the spatial organization of cell mosaics is disrupted during regeneration. The results suggest that retinal regeneration may represent a spatially disrupted recapitulation of retinal developmental mechanisms.

Focal hippocampal gain of NGF function elicits specific septal cholinergic reorganization.

The influence of NGF on the neuroanatomic substrate(s) subserving learning and memory was probed by somatic mosaic analysis. NGF XAT mice underwent unilateral hippocampal delivery of virus vector expressing cre recombinase to produce focal NGF gain of function mosaics. Activated mice expressing increased NGF, but not control mice transduced with the lacZ gene showed reorganization of the septohippocampal projection assessed by retrograde labeling with Fluorogold (FG). Mice mosaic for NGF function demonstrated in ipsilateral medial septum and diagonal band significant increases in FG-labeled cell bodies (94%), ChAT-labeled cells (55%), double-positive cells (190%) and increased somal size of double-positive cells (56%) than did non-activated mice. These results indicate that reorganization of the cholinergic septal input to a specific hippocampal region is promoted by gain of NGF function.

Altered glutamate uptake in peripheral tissues from Down syndrome patients.

Overexpression of APP and SOD induces beta-amyloid deposition and oxidative stress in Down syndrome (DS) patients. Both phenomena may impair glutamate transport and decreased glutamate uptake sites have been demonstrated in patient brains at autopsy. Since alterations of APP metabolism and oxidative damage are systemic, we investigated glutamate uptake in platelets and fibroblasts from DS patients to explore whether abnormalities in this process are inherent properties of DS cells and not secondary to neurodegeneration. Glutamate uptake was significantly decreased in platelets (P<0.005 vs. control) and fibroblasts (P<0.001 vs. control) from DS patients, particularly in those with free trisomy and with mitochondrial point mutations. Systemic impairment of glutamate uptake in DS is suggested, probably related to APP overexpression and mitochondrial dysfunction. Such mechanisms may contribute to neurodegeneration and dementia development in these patients.

Bilateral frontal polymicrogyria and epilepsy in a patient with Turner mosaicism: a case report.

Turner's syndrome (TS) is rarely associated with serious abnormalities of brain structure or malformations of cortical development. We report a 17-year-old girl with TS and 45,XO/46,XX mosaicism presenting bilateral frontal polymicrogyria (BFP) and epilepsy. To our knowledge, the association between TS and BFP has never been reported to date. Our observation confirms that in humans the X-chromosome plays an important role in the development and specialization of brain structure and function. We hypothesize that the absence or abnormalities of developmental genes localized on the X-chromosome could be involved in the pathogenesis of BFP observed in our patient.

Differential pattern in tissue-specific somatic mosaicism of expanded CAG trinucleotide repeats in dentatorubral-pallidoluysian atrophy, Machado-Joseph disease, and X-linked recessive spinal and bulbar muscular atrophy.

We investigated the somatic mosaicism of trinucleotide repeat expansion in the neural and nonneural tissues of a dentatorubral-pallidoluysian atrophy (DRPLA), Machado-Joseph disease (MJD), and spinal and bulbar muscular atrophy (SBMA) patient and their correlation to the topographical distribution of the pathological involvement. The spatial pattern of tissue-specific somatic mosaicism in the CAG repeat size was significantly different among the DRPLA, MJD and SBMA patients. The size of the major bands of the mutant CAG repeat allele was significantly smaller in the cerebellar cortex in both DRPLA and MJD patients by 6 and 2 repeat units respectively and larger in the colon and liver of DRPLA by 5 repeats or more. There were also 1-2 repeat-sized small variations of major band size among the neural tissues in DRPLA. In contrast, there was no tissue-specific variation of major bands of CAG repeats and diversity of extra bands among the examined tissues including the cerebellum in the SBMA patient. There was no parallel occurrence of tissue-specific CAG instability and severity of neuropathological involvement in the neural and nonneural tissues of DRPLA, MJD and SBMA patients. Lack of significant tissue-specific somatic mosaicism in SBMA including the cerebellar cortex may suggest that CAG repeat expansion in the mutant androgen receptor gene is far more stable compared with that in DRPLA and MJD as well as those reported in Huntington's disease.

Single cell analysis of CAG repeat in brains of dentatorubral-pallidoluysian atrophy (DRPLA).

Somatic mosaicism of an expanded repeat is present in tissues of patients with triplet repeat diseases. Of the spinocerebellar ataxias associated with triplet repeat expansion, the most prominent heterogeneity of the expanded repeat is seen in dentatorubral-pallidoluysian atrophy (DRPLA). The common feature of this somatic mosaicism is the difference in the repeat numbers found in the cerebellum as compared to other tissues. The expanded allele in the cerebellum shows a smaller degree of expansion. We previously showed by microdissection analysis that the expanded allele in the granular layer in DRPLA cerebellum has less expansion than expanded alleles in the molecular layer and white matter. Whether this feature of lesser expansion in granule cells is common to other types of neurons is yet to be clarified. We used a newly developed excimer laser microdissection system to analyze somatic mosaicism in the brains of two patients, one with early- and another with late-onset DRPLA, and used single cell PCR to observe the cell-to-cell differences in repeat numbers. In the late onset patient, repeat expansion was more prominent in Purkinje cells than in granule cells, but less than that in the glial cells. In the early onset patient, repeat expansion in Purkinje cells was greater than in granule cells but did not differ from that in glial cells. These findings suggest that there is a difference in repeat expansion among neuronal subgroups and that the number of cell division cycles is not the only determinant of somatic mosaicism.

The role of X-chromosome inactivation in the manifestation of Rett syndrome.

X-chromosome inactivation (XCI) is random in the majority of patients with classical Rett syndrome (RTT). Preferential inactivation of the X chromosome with the mutated MECP2 gene is found in mildly symptomatic or asymptomatic carrier females. These findings lead to a hypothesis that random XCI is causally involved in the pathogenesis of RTT in heterozygous females. It is the cluster of functionally defective nerve cells lacking fully functional MeCP2 generated by inactivation of normal MECP2 allele that causes the wide spectrum of RTT symptoms. Thus, RTT is a rare human disease manifestation which is triggered most probably by random XCI.

Pallister-Mosaic syndrome and neuronal migration disorder.

We diagnosed Pallister-Mosaic syndrome (PMS) in a 4-month-old female infant. In addition to the presence of non-specific anomalies, involving anorectal, finger and ear anomalies, characteristic cranio-facial features and irregular skin lesions that appeared after age 2 months suggested the possibility of genetic mosaicism, PMS in particular. Fluorescence in situ hybridization technique revealed an extra copy of chromosome 12p; i (12p) in 30% of cultured skin fibroblasts. When focal skin lesions accompany neurodevelopmental disabilities in early infancy, genetic analysis for mosaicism should be considered for differential diagnosis. Significantly, we describe several phenotypic features and neuroimaging findings of the PMS in the present case, which have not been described in previous reports. The neuroimaging abnormalities we encountered, such as polymicrogyria, speculating congenital brain anomaly, may explain the severe motor and intellectual disabilities of PMS.

Ring chromosome 14 complicated with complex partial seizures and hypoplastic corpus callosum.

A Japanese male with mosaicism of ring chromosome 14 and chromosome 14 monosomy is described. He demonstrated the characteristic morphologic features of ring chromosome 14, in addition to mental retardation and epileptic seizures. Clusters of complex partial seizures, one of which originated in the left frontocentral region on electroencephalographic monitoring, were evident. His seizures responded to phenobarbital, and his mental and motor development was only mildly retarded. Magnetic resonance imaging revealed a hypoplastic corpus callosum, previously unknown in association with this syndrome.

Disappearing trisomy 8 mosaicism.

A case is presented of a patient with disappearing trisomy 8 mosaicism initially thought to have stigmata of the fragile X syndrome. This case is interesting for two reasons. First, it demonstrates the occurrence of "disappearing mosaicism," a phenomenon first described by LaMarche et al, in 1967. Our patient, initially studied in 1991 by two laboratories and found to be mosaic for chromosome 8 trisomy, was apparently normal by both GTG-banding and fluorescent in situ hybridization (FISH) when studied in 1996. Second, this case further underscores the fact that except under special circumstances, it is important that GTG-banding analysis be performed so that the entire human genome be examined in addition to scoring for the fragile X mutation on Xq27.3. In a recent review of the existing database at Rhode Island Hospital on chromosomal abnormalities found in patients referred because of a question of the fragile X syndrome during the period from January 1, 1990 to June 30, 1995, it was found that the frequency of other chromosomal abnormalities in patients referred because of a question of fragile X syndrome equaled or exceeded that of patients found to be positive for fragile X. Our figures, consistent with those reported in the literature, underscore the value of routine karyotyping in this population of patients.

Large retinal ganglion cells that form independent, regular mosaics in the ranid frogs Rana esculenta and Rana pipiens.

Population-based studies of ganglion cells in retinal flatmounts have helped to reveal some of their natural types in mammals, teleost fish and, recently, the aquatic mesobatrachian frog Xenopus laevis. Here, ganglion cells of the semiterrestrial neobatrachian frogs Rana esculenta and Rana pipiens have been studied similarly. Ganglion cells with large somata and thick dendrites could again be divided into three mosaic-forming types with distinctive stratification patterns. Cell dimensions correlated inversely with density, being smallest in the visual streak. Cells of the alpha a mosaic (< 0.2% of all ganglion cells) had the largest somata at each location (often displaced) and their trees were confined to one shallow plane within sublamina a of the inner plexiform layer. In regions of high regularity, many trees were symmetric. Elsewhere, asymmetric, irregular trees predominated and their dendrites, although sparsely branched, achieved consistent coverage by intersecting in complex ways. Cells of the alpha ab mosaic were more numerous (approximately 0.7%) and had large somata, smaller (but still large) trees, and dendrites that branched extensively in two separate shallow planes in sublaminae a and b. The subtrees did not always match in symmetry, and each subtree tessellated independently with its neighbors. Cells of the alpha c mosaic (approximately 0.1%) had large, orthotopic somata and large, sparse trees (often asymmetric and irregular) close to the ganglion cell layer. Nearest-neighbor analyses and spatial correlograms confirmed that each mosaic was regular and independent. Densities, proportions, sizes, and mosaic statistics are tabulated for all three types, which are compared with types defined by size and symmetry in R. pipiens, by discriminant analysis in R. temporaria, by physiological response in both, and by mosaic analysis in Xenopus and several teleosts. The variable stratification of these otherwise similar types across species is consistent with other evidence that stratification may be determined, in part, by functional interactions.

The effect of confined placental mosaicism on development of the human aneuploid conceptus.

The rate of meiotic errors in gametogenesis is high, resulting in approximately 30% of cleaving embryos with chromosomal defect. In addition, mitotic errors occur commonly in chromosomally normal and abnormal embryos. The most likely outcome of the mitotic error is CPM. Mitotic mutation in trisomic blastocysts may lead to diploid/trisomic trophoblast with intrauterine survival of the trisomic fetus or may result in a diploid fetus supported by a trisomic placenta. The outcome of pregnancies with a trisomic placenta and diploid fetus largely depends on the specific chromosome involved and on the presence or absence of uniparental disomy in the fetus.

Prenatal diagnosis and dysmorphic findings in mosaic trisomy 16.

We report two cases of mosaic trisomy 16 diagnosed by amniocentesis, with dysmorphic findings in both cases evident upon delivery. Following elective termination, case 1 demonstrated a trisomy 16 cell line in fetal skin (4 per cent) and placental tissue (64 per cent). Molecular studies on the disomic cell line indicated that both chromosome 16s were maternal in origin, suggesting loss of the paternal chromosome 16 from a trisomic zygote (uniparental heterodisomy). At birth, case 2 demonstrated only disomic cells in skin and blood, with trisomy 16 present in 4 per cent of cells from the amnion. Molecular studies confirmed both maternal and paternal contributions of the chromosome 16s. We analysed DNA from one previously reported case of mosaic trisomy 16 (Williams et al., 1992) and failed to find signs of uniparental disomy in this child with congenital heart defects. These cases had distinctive but different dysmorphic features. We suggest that trisomy 16 embryos may revert to disomy during the course of pregnancy, allowing for longer survival with various abnormalities in growth and morphogenesis. The clinical significance of prenatally detected mosaic trisomy 16 may not be completely defined by additional cytogenetic, molecular, and ultrasound studies.

Tandem duplication mosaicism: characterization of a mosaic dup(5q) and review.

Mosaicism for tandem duplications is rare. Most patients reported had abnormal phenotypes of varying severity, depending on the chromosomal imbalance involved and the level of mosaicism. Post-zygotic unequal sister-chromatid exchange has been proposed as the main mechanism for tandem duplication mosaicism. However, previous molecular analyses have implicated both meiotic and post-zygotic origins for the duplication. We describe a newborn male who was originally diagnosed in utero with arrhythmia and tetralogy of Fallot. He had multiple dysmorphic features including telecanthus, blepharophimosis, high broad nasal bridge with a square-shaped nose, flat philtrum, thin upper lip, down-turned corners of the mouth, high-arched palate, micrognathia, asymmetric ears, and long, thin fingers and toes. Karyotyping of peripheral blood lymphocytes showed mosaicism for a tandem duplication of part of the long arm of one chromosome 5: mos46,XY,dup(5)(q13q33)[6]/46,XY[45]. Fibroblast cultures had the same mosaic karyotype with a higher frequency of the dup(5) clone: mos46,XY,dup(5)(q13q33)[9]/46,XY[21]. Fluorescence in situ hybridization analysis with a wcp5 confirmed the chromosome 5 origin of the additional material. Parental karyotypes were normal indicating a de novo origin of the dup(5) in the proband. Molecular analyses of chromosome 5 sequence-tagged-site (STS) markers in our family were consistent with a post-zygotic origin for the duplication. Therefore, mosaicism for tandem duplications can arise both through meiotic or mitotic errors, as a result of unequal crossing over or unequal sister-chromatid exchange, respectively. Our review indicates that mosaicism for tandem duplications is likely under-ascertained and that parental karyotyping of probands with non-mosaic tandem duplications should be performed.

Molecular genetic aspects of the phakomatoses: tuberous sclerosis complex and neurofibromatosis 1.

The phakomatoses are a diverse group of diseases characterized by skin lesions in early childhood followed by the development of tumors in many other organs. Tuberous sclerosis complex and neurofibromatosis 1 are of special interest to the neurologist because of their prominent neuro-oncological and neuro-developmental consequences. The cloning of genes responsible for these two diseases has led to the identification of causative mutations, an understanding of basic cellular pathophysiology and the development of animal models. Current laboratory investigations are focused on bringing clinical relevance to these findings, including the prospects of molecular diagnostics and rational therapeutics.

Mushroom bodies are not required for courtship behavior by normal and sexually mosaic Drosophila.

To elucidate the effect of feminization of male Drosophila brain cells on courtship control, we performed a large scale screening of expression drivers that can suppress male-specific behavior with transformer gene expression. Two drivers caused essentially total courtship suppression. The expression pattern of these drivers did not show any correlation with the mushroom bodies or the antennal lobes, the regions that have been suggested to play important roles in courtship. Ablation of mushroom bodies using hydroxyurea treatment did not affect this courtship suppression. The ablation did not change either wild-type heterosexual behavior or bisexual behavior caused by transformer expression driven by the same drivers used in the previous studies to suggest the involvement of the mushroom bodies in courtship. Our results show that feminization of different nonoverlapping cells in other parts of the protocerebrum was sufficient to cause the same bisexual or suppressed-courtship phenotype. Thus, contrary to previous assumptions, the mushroom bodies are not required for the control of courtship. Present evidence supports its mediation by other distributed protocerebral regions.