A study of FHL1, BAG3, MATR3, PTRF and TCAP in Australian muscular dystrophy patients

FHL1, BAG3, MATR3 and PTRF are recently identified myopathy genes associated with phenotypes that overlap muscular dystrophy. TCAP is a rare reported cause of muscular dystrophy not routinely screened in most centres. We hypothesised that these genes may account for patients with undiagnosed forms of muscular dystrophy in Australia. We screened a large cohort of muscular dystrophy patients for abnormalities in FHL1 (n = 102) and TCAP (n = 100) and selected patients whose clinical features overlapped the phenotypes previously described for BAG3 (n = 9), MATR3 (n = 15) and PTRF (n = 7). We found one FHL1 mutation (c.311G>A, p.C104Y) in a boy with rapidly progressive muscle weakness and reducing body myopathy who was initially diagnosed with muscular dystrophy. We identified no pathogenic mutations in BAG3, MATR3, PTRF or TCAP. In conclusion, we have excluded these five genes as common causes of muscular dystrophy in Australia. Patients with reducing body myopathy may be initially diagnosed as muscular dystrophy.     

Linkage disequilibrium and haplotype tagging polymorphisms in the<Emphasis Type="Italic"> Tau</Emphasis> H1 haplotype

We and others have previously detected association of the Tau H1 haplotype on chromosome 17 with risk of idiopathic Parkinson disease (PD). The H1 haplotype appears to have a fundamental importance in neurodegeneration, as multiple studies have shown it is also associated with an increased risk for progressive supranuclear palsy, corticobasal degeneration, frontotemporal lobar degeneration syndromes, and primary progressive aphasia. Therefore, to divide the H1 haplotype into sub-haplotypes that could be more significantly associated with the risk of developing PD, and to delimit the genes lying in the H1 haplotype, we analyzed 34 single nucleotide polymorphisms (SNPs) spanning over 3.15 megabases in the region containing Tau. These SNPs are located in or flank the corticotropin-releasing hormone receptor 1, presenilin homolog 2, Tau, Saitohin, and KIAA1267 genes. Analysis of linkage disequilibrium (LD) using these 34 SNPs suggests that the H1 haplotype extends over about 1.3 megabases, making it the largest region of LD reported to date. Of the 29 SNPs lying in this region of LD, 5 were identified as haplotype tagging SNPs (htSNPs), capturing 96% of the samples haplotype diversity. Association analysis with these htSNPs revealed a new H1 sub-haplotype that is significantly associated with PD (P<0.02). These results define the genes and regulatory regions included in this region of LD, containing an important susceptibility allele contributing to increased risk of neurodegeneration.     

Study of Survival of Motor Neuron (SMN) and Neuronal Apoptosis Inhibitory Protein (NAIP) gene deletions in SMA patients

In view of the paucity of deletion studies of survival of motor neuron (SMN) and neuronal apoptosis inhibitor protein (NAIP) genes in Indian SMA patients, this study has been undertaken to determine the status of SMN1, SMN2 and NAIP gene deletions in Indian SMA patients. Clinically and neurophysiologically diagnosed SMA patients were included in the study. A gene deletion study was carried out in 45 proximal SMA patients and 50 controls of the same ethnic group. Both SMN1 and NAIP genes showed homozygous absence in 76 % and 31 % respectively in proximal SMA patients. It is proposed that the lower deletion frequency of SMN1 gene in Indian patients may be due to mutations present in other genes or population variation, which need further study.     

Association study of cholesterol-related genes in Alzheimer’s disease

Alzheimer’s disease (AD) is a genetically complex disorder, and several genes related to cholesterol metabolism have been reported to contribute to AD risk. To identify further AD susceptibility genes, we have screened genes that map to chromosomal regions with high logarithm of the odds scores for AD in full genome scans and are related to cholesterol metabolism. In a European screening sample of 115 sporadic AD patients and 191 healthy control subjects, we analyzed single nucleotide polymorphisms in 28 cholesterol-related genes for association with AD. The genes HMGCS2, FDPS, RAFTLIN, ACAD8, NPC2, and ABCG1 were associated with AD at a significance level of P ≤ 0.05 in this sample. Replication trials in five independent European samples detected associations of variants within HMGCS2, FDPS, NPC2, or ABCG1 with AD in some samples (P = 0.05 to P = 0.005). We did not identify a marker that was significantly associated with AD in the pooled sample (n = 2864). Stratification of this sample revealed an APOE-dependent association of HMGCS2 with AD (P = 0.004). We conclude that genetic variants investigated in this study may be associated with a moderate modification of the risk for AD in some samples. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The chick/quail transplantation model: Discovery of the isthmic organizer center

This paper summarizes chick/quail transplantation experiments performed in the INSERM U106 by Alvarado-Mallart's group from 1989 to 2002. First, it will present the various steps leading us to demonstrate that, at stage 10 of Hamburger and Hamilton, the avian neuroepithelium is still competent to change its fate influenced by environmental inductive factors and that these factors emanate from the cerebellar neuroepithelium; then, it will be briefly reported, experiments aimed to characterize the genetic cascade involved in the formation of the midbrain/hindbrain boundary and the specification of the meso-isthmic-cerebellar domain.     

Lrrk2 G2019S substitution in frontotemporal lobar degeneration with ubiquitin-immunoreactive neuronal inclusions

Leucine-rich repeat kinase 2 (LRRK2) mutation carriers can develop clinical symptoms other than typical parkinsonism such as dementia, amyotrophy or dystonia. To determine if LRRK2 mutations might be involved in frontotemporal dementia (FTD), 5 individuals with multiplex familial FTD kindreds and 41 pathologically confirmed cases of FTD, including 23 with a family history of dementia, were screened for genetic variations in the LRRK2 gene. We identified a LRRK2 mutation leading to the G2019S amino acid substitution in a 79-year-old woman with frontotemporal lobar degeneration with ubiquitinated neuronal intranuclear inclusions (FTLD-U/NII) and a possible family history of tremor. These findings may be coincidental; however, there is a small nucleus of LRRK2-positive patients displaying atypical features suggesting a role for this protein in other neurodegenerative disorders.     

Granulomatous amebic encephalitis: a review and report of a spontaneous case from Venezuela

Granulomatous amebic encephalitis (GAE), or meningoencephalitis due to Acanthamoeba spp. and leptomyxid ameba are uncommon CNS infections that generally occur in immunocompromised hosts. We describe a case of GAE caused by Balamuthia mandrillaris previously designated as a leptomyxid ameba, in an apparently healthy 14-year-old Venezuelan boy. This case was characterized by sudden onset of seizures, focal neurologic signs and by a prolonged clinical course (from November 1992 to March 1993). Neuroimaging studies showed cerebral hypodense lesions in cerebral hemispheres, brain stem and cerebellum. Microscopically, we found a chronic granulomatous inflammatory reaction with necrotizing angiitis, large numbers of amebic trophozoites and few cysts in perivascular spaces and within necrotic CNS tissue. The amebas were identified as B. mandrillaris based on their immunofluorescence reactivity with the anti-B. mandrillaris serum. So far, 30 cases of GAE due to B. mandrillaris have been recognized in humans, two in AIDS patients. No visceral involvement by free-living amebas or any other significant abnormality was observed. This patient developed spontaneous GAE, but it remains possible that an undiagnosed abnormality in cell-mediated immunity or a deficient humoral immune response may explain the susceptibility of this patient to this opportunistic infection.Supported in part by the Pathology Education and Research Foundation (PERF) of the Department of Pathology, University of Pittsburgh     

Combined molecular analysis of BRAF and IDH1 distinguishes pilocytic astrocytoma from diffuse astrocytoma

Separation of pilocytic astrocytoma from diffuse astrocytomas frequently poses problems mostly related to small sample size. Precise classification and grading are essential due to different therapeutic strategies prompted by diagnoses of pilocytic astrocytoma WHO grade I, diffuse astrocytomas WHO grade II or anaplastic astrocytoma WHO grade III. Recently, genomic aberrations with a high specificity for distinct glioma entities have been described. Pilocytic astrocytomas carry a duplication at chromosome band 7q34 containing a BRAF–KIAA1549 gene fusion in the majority of cases. IDH1 mutations are observed very frequently in adult astrocytomas and IDH2 mutations have been reported in some astrocytomas. We examined a series of 120 astrocytomas including 70 pilocytic astrocytomas WHO grade I and 50 diffuse astrocytomas WHO grade II for both, BRAF–KIAA1549 fusion with a newly developed FISH assay and mutations in IDH1 and IDH2 by direct sequencing. Pilocytic astrocytomas contained the BRAF fusion in 49 cases (70%) but neither IDH1 nor IDH2 mutations. Astrocytomas WHO grade II exhibited IDH1 mutations in 38 cases (76%) but neither IDH2 mutations nor BRAF fusions. Thus, combined molecular analysis of BRAF and IDH1 is a sensitive and highly specific approach to separate pilocytic astrocytoma from diffuse astrocytoma.     

Behavioral transformations during metamorphosis: remodeling of neural and motor systems

During insect metamorphosis, neural and motor systems are remodeled to accommodate behavioral transformations. Nerve and muscle cells that are required for larval behavior, such as crawling, feeding and ecdysis, must either be replaced or respecified to allow adult emergence, walking, flight, mating and egg-laying. This review describes the types of cellular changes that occur during metamorphosis, as well as recent attempts to understand how they are related to behavioral changes and how they are regulated. Within the periphery, many larval muscles degenerate at the onset of metamorphosis and are replaced by adult muscles, which are derived from myoblasts and, in some cases, remnants of the larval muscle fibers. The terminal processes of many larval motoneurons persist within the periphery and are essential for the formation of adult muscle fibers. Although most adult sensory neurons are born postembryonically, a subset of larval proprioceptive neurons persist to participate in adult behavior. Within the central nervous system, larval neurons that will no longer be necessary die and some adult interneurons are born postembryonically. By contrast, all of the adult motoneurons, as well as some interneurons and modulatory neurons, are persistent larval cells. In accordance with their new behavioral roles, these neurons undergo striking changes in dendritic morphology, intrinsic biophysical properties, and synaptic interactions.     

Basal ganglia calcifications and ALS syndrome

We report the case of a patient with idiopathic hypoparathyroidism and unusually large symmetrical calcifications in the basal ganglia, thalami, cerebellar hemispheres and brainstem, who clinically presented an ALS-like syndrome. We discuss the possible role of abnormal calcium metabolism in the pathogenesis of motoneuron disease.

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Sommario Riportiamo un caso di ipoparatiroidismo idiopatico con calcificazioni estese e simmetriche nei gangli della base, talamo, emisferi cerebellari, tronco encefalico, e che presentava clinicamente una sindrome tipo SLA. Sulla base dei dati riportati dalla letteratura vengono quindi discussi ed ipotizzati gli eventuali rapporti patogenetici fra anormalità del metabolismo del calcio e malattia del motoneurone.

     

Role of Peripheral Adenosine A1 Receptors in the Regulation of Sleep Apneas in Rats

The effects of administration of N⁶-p-sulfophenyladenosine (p-SPA), a peripheral adenosine A₁ receptor agonist, and 8-(p-sulfophenyl)theophylline (p-SPT), a peripheral adenosine A₁ receptor blocker, on spontaneous apneas were studied in 10 adult Sprague–Dawley rats by monitoring respiration, sleep, and blood pressure for 6 h. Intraperitoneal injection of p-SPA (1 mg/kg) to rats suppressed spontaneous central apneas during non-rapid eye movement sleep by 50% in comparison to control recordings (p = 0.03). This effect was blocked by pretreatment with an equimolar dose of p-SPT (0.67 mg/kg) indicating that p-SPA suppression of apneas was receptor mediated in the peripheral nervous system. Administration of p-SPA did not affect apnea expression in rapid eye movement sleep and had no effect on sleep or blood pressure at the dose tested. Administration of p-SPT (0.67, 6.7, and 30 mg/kg) to rats had no effect on apneas, sleep, or blood pressure. The lack of p-SPT effect on sleep apneas argues against a physiologic role for endogenous adenosine in the peripheral nervous system as a modulator of sleep apnea expression under baseline conditions.     

Association studies of variants in MEIS1, BTBD9, and MAP2K5/SKOR1 with restless legs syndrome in a US population

Background

A genome-wide association study (GWAS) identified significant association between variants in MEIS1, BTBD9, and MAP2K5/SKOR1 and restless legs syndrome (RLS). However, many independent replication studies are needed to unequivocally establish a valid genotype-phenotype association across various populations. To further validate the GWAS findings, we investigated three variants, rs2300478 in MEIS1, rs9357271 in BTBD9, and rs1026732 in MAP2K5/SKOR1 in 38 RLS families and 189 RLS patients/560 controls from the US for their association with RLS.

Method

Both family-based and population-based case-control association studies were carried out.

Results

The family-based study showed that SNP rs1026732 in MAP2K5/SKOR1 was significantly associated with RLS (P = 0.01). Case-control association studies showed significant association between all three variants and RLS (P = 0.0001/OR = 1.65, P = 0.0021/OR = 1.59, and P = 0.0011/OR = 1.55 for rs2300478, rs9357271, and rs1026732, respectively).

Conclusion

Variants in MEIS1, BTBD9, and MAP2K5/SKOR1 confer a significant risk of RLS in a US population.     

Dopamine D2 receptor antagonists induce immediate early genes in the rat striatum

The acute effects of dopamine D₂ antagonists and agonists on the expression of c-jun, zif-268, ETR101 and c-fos in the rat striatum were studied. A single injection IP of haloperidol (2 mg/kg) or sulpiride (100 mg/kg) produced a rapid and transient increase in c-jun, zif-268 and c-fos mRNA. ETR101 was not activated. These inductions were dose dependent and were specifically blocked by pretreatment with a D₂ agonist (1 mg/kg quinelorane). Quinelorane alone had no effect. Thus, dopamine D₂ receptors inhibit the expression of a particular set of immediate early genes in the striatum.     

Association of dopamine transporter and monoamine oxidase molecular polymorphisms with sudden infant death syndrome and stillbirth: new insights into the serotonin hypothesis

Recent findings demonstrated the role of neurotransmitters in the aetiopathogenesis of sudden unexpected deaths in infancy. Although genes involved in serotonin metabolism have been proposed as risk factors for sudden infant death syndrome (SIDS), the contribution of additional neurotransmitters and genes different from the serotonin transporter (SLC6A4, 5-HTT) has not been investigated. Considering the common metabolic pathway and synergism between dopamine and serotonin, the role of dopamine transporter (SLC6A3, DAT) and monoamine oxidase A (MAOA) genes in SIDS and stillbirth (sudden intrauterine unexplained death, SIUD) was investigated. Genotypes and allelic frequencies of DAT and MAOA were determined in 20 SIDS and five stillbirth cases and compared with 150 controls. No association was found between DAT polymorphisms and SIDS either at genotype (P = 0.64) or allelic (P = 0.86) level; however, a highly significant association was found between MAOA genotypes (P = 0.047) and alleles (P = 0.002) regulating different expression patterns (3R/3R vs 3.5R/3.5R + 4R/4R) in SIDS + SIUD and controls. Analysis of combined 5-HTTLPR (serotonin transporter linked polymorphic region)/MAOA genotypes revealed that frequency of L/L-4R/4R genotype combination was eightfold higher in SIDS + SIUD than in controls (P < 0.001). Findings are discussed considering the metabolic association among DAT, 5-HTT and MAOA with special emphasis on the linked action of 5-HTT/MAOA in regulating serotonin metabolism of SIDS and SIUD infants.     

Activation of neuroblast proliferation in explant culture of the Drosophila larval CNS

The developmental control of neuroblast proliferation is absolutely required for the assembly and function of the central nervous system. A lethal mutation in trol results in the failure of quiescent neuroblasts to begin division at the appropriate time. I have established a culture system in which quiescent neuroblasts in explants of Drosophila larval CNSs initiate cell division in vitro to normal in vivo levels. This activation requires removal of the CNS for culture after a specific developmental stage and the presence of fetal calf serum or a larval extract in the medium. Either supplement is effective when heat-treated. Substitution of the steroid hormone ecdysone or the non-steroidal ecdysone analog RH5992 for either fetal calf serum or larval extract also results in activation of neuroblast proliferation. Culture of trol^sd CNSs with wildtype larval extract or ecdysone results in the defective neuroblast proliferation phenotype observed in trol mutants in vivo, while culture of wildtype CNSs with trol^sd extract produces normal neuroblast proliferation.     

Mutation analysis of the Ras pathway genes<Emphasis Type="Italic"> NRAS</Emphasis>,<Emphasis Type="Italic"> HRAS</Emphasis>,<Emphasis Type="Italic"> KRAS</Emphasis> and<Emphasis Type="Italic"> BRAF</Emphasis> in glioblastomas

Aberrant activation of Ras signaling is a common finding in human glioblastomas. To determine the contribution of Ras gene mutations to this aberration, we screened 94 glioblastomas for mutations in the three Ras family genes NRAS, KRAS and HRAS. All tumors were additionally analyzed for mutations in BRAF, which encodes a Ras-regulated serine/threonine kinase with oncogenic properties. Mutation analysis of the entire coding regions of NRAS and KRAS, as well as the known mutation hot-spot sites in HRAS, identified somatic point mutations in two glioblastomas, both affecting codon 12 of NRAS (c.35G>A, p.G12D). Three additional tumors carried BRAF mutations altering the known hot-spot codon 599 (c.1796T>A, p.V599E). None of these five glioblastomas showed amplification of the EGFR or PDGFRA genes, while three of the tumors, including two with NRAS and one with BRAF mutation, demonstrated PTEN missense mutations or loss of PTEN mRNA expression. Taken together, our data suggest activating mutations in NRAS or BRAF as a molecular alteration that contributes to aberrant Ras signaling in a small fraction of glioblastomas.