Fine mapping of a dravet syndrome modifier locus on mouse chromosome 5 and candidate gene analysis by rna-seq database 3 normal forms
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data recovery utility A substantial number of mutations have been identified in voltage-gated sodium channel genes that result in various forms of human epilepsy. Data recovery usb flash drive SCN1A mutations result in a spectrum of severity ranging from mild febrile seizures to Dravet syndrome, an infant-onset epileptic encephalopathy. Data recovery uk Dravet syndrome patients experience multiple seizures types that are often refractory to treatment, developmental delays, and elevated risk for SUDEP. Data recovery usb stick The same sodium channel mutation can produce epilepsy phenotypes of varying clinical severity. Data recovery using linux This suggests that other factors, including genetic, modify the primary mutation and change disease severity.
Data recovery usa Mouse models provide a useful tool in studying the genetic basis of epilepsy. Data recovery utah The mouse strain background can alter phenotype severity, supporting a contribution of genetic modifiers in epilepsy. Ease use data recovery The Scn1a +/- mouse model has a strain-dependent epilepsy phenotype. Data recovery vancouver Scn1a +/- mice on the 129S6/SvEvTac (129) strain have a normal phenotype and lifespan, while [129xC57BL/6J]F1- Scn1a +/- mice experience spontaneous seizures, hyperthermia-induced seizures and high rates of premature death. Data recovery video We hypothesize the phenotypic differences are due to strain-specific genetic modifiers that influence expressivity of the Scn1a +/- phenotype. Data recovery victoria bc Low resolution mapping of Scn1a +/- identified several Dravet syndrome modifier ( Dsm) loci responsible for the strain-dependent difference in survival. Data recovery vancouver bc One locus of interest, Dsm1 located on chromosome 5, was fine mapped to a 9 Mb region using interval specific congenics. Data recovery windows RNA-Seq was then utilized to identify candidate modifier genes within this narrowed region. Data recovery wizard free Three genes with significant total gene expression differences between 129S6/SvEvTac and [129xC57BL/6J]F1 were identified, including the GABA A receptor subunit, Gabra2. Data recovery wizard professional Further analysis of Gabra2 demonstrated allele-specific expression. Data recovery windows 10 Pharmological manipulation by clobazam, a common anticonvulsant with preferential affinity for the GABRA2 receptor, revealed dose-dependent protection against hyperthermia-induced seizures in Scn1a +/- mice. Data recovery western digital These findings support Gabra2 as a genetic modifier of the Scn1a +/- mouse model of Dravet syndrome. Data recovery wd passport A significant improvement in survival was observed with lines ISC-A (4–118 Mb; p<0.01), ISC-B (4–74.9 Mb; p<0.0001), ISC-C (53–118 Mb; p<0.05) and ISC-D (64.6–74.9 Mb; p<0.0002) ( Fig 1B). Data recovery wizard for mac The strongest effect on survival was observed in mice carrying homozygous 129/129 alleles in ISC-B and ISC-D, with 8 week survival of 91.3% and 92.8%, respectively, compared to only 28.5% in heterozygous F1. Data recovery windows 7 Scn1a +/- control mice ( Fig 1B). Data recovery xfs ISC-D carried the smallest 129-derived segment (64.6–73.9 Mb) and refined the map interval to ~9 Mb ( Fig 1A). Os x data recovery We next addressed whether the survival advantage conferred by 129 alleles in the ISC-D interval correlated with a decreased seizure burden. Os x data recovery software F1. Moto x data recovery Scn1a +/-offspring carrying either homozygous (129/129) or control heterozygous (129/B6) alleles in ISC-D were monitored from postnatal day 21 (P21) through P24 to determine the frequency of generalized tonic-clonic (GTC) seizures. Os x data recovery free Scn1a +/- mice carrying homozygous (129/129) alleles in the ISC-D interval did not have a significantly reduced seizure frequency compared to heterozygous F1. Data recovery yelp Scn1a +/- control mice (ISC-D: 0.024 ± 0.015 seizures/hr; F1 control: 0.022 ± 0.02 seizures/hr). Data recovery youtube Dissociation of the survival and seizure frequency phenotypes was somewhat surprising, and raised the question of whether seizure frequency and survival are correlated in F1.Scn1a +/- mice. Data recovery zagreb To address this question, we examined a separate phenotyping cohort and determined seizure counts for F1. Sony xperia z data recovery Scn1a +/- mice between P19 to P24. Z a r data recovery In F1. Raid 0 data recovery Scn1a +/- mice experiencing seizures, total numbers of seizures were compared with survival throughout the monitoring period (Survivors: 7.95 ± 1.42 seizures; Lethals: 5.13 ± 1.55 seizures). Raid 0 data recovery software This comparison revealed that high seizure frequency did not correlate with premature lethality in F1. Raid 1 data recovery Scn1a +/- (Pearson r 2 = 0.01, p>0.8111), consistent with our observations with ISC-D. No 1 data recovery software We further evaluated forebrain Gabra2 transcript and protein expression in ISC-D mice carrying homozygous 129/129, heterozygous 129/B6 or homozygous B6/B6 alleles in Dsm1. 1 care data recovery software We used digital droplet RT-PCR (ddRT-PCR) to evaluate transcript expression of Gabra2, as well as expression of other GABA A receptor genes located within Dsm1, including Gabra4, Gabrb1 and Gabrg1. Raid 1 data recovery software Gabra2 expression was significantly different between all genotypes (F 2,15 = 206.4, p<0.0001). Data recovery 2016 We observed the highest levels of Gabra2 transcript with homozygosity for 129 alleles in Dsm1, intermediate levels with heterozygosity, and the lowest levels with B6 homozygosity ( Fig 3A). Ipad 2 data recovery Transcript expression of the other Gabr genes in the interval did not differ between genotypes ( S1 Fig). Note 2 data recovery Relative GABRA2 protein expression determined by immunoblotting correlated with the transcript levels, showing the highest expression with homozygous 129 alleles in Dsm1, intermediate with heterozygous alleles, and lowest with B6 homozygosity ( Fig 3B and 3C). Note 3 data recovery These results demonstrate that the level of Gabra2 expression is regulated by alleles within the Dsm1 interval, and that high expression of Gabra2 is correlated with the protective allele for Scn1a +/- survival. Level 3 data recovery We used clobazam, an anticonvulsant drug that has preferential affinity for the GABRA2 subunit [ 17– 19], as a pharmacological tool to further evaluate Gabra2 as a candidate modifier gene. Data recovery iphone 4s Both Dravet syndrome patients and F1. Iphone 4 data recovery Scn1a +/- Dravet mice experience seizures triggered by hyperthermia. Iphone 4 data recovery software We evaluated the effect of clobazam on survival and hyperthermia seizure threshold in F1. Nexus 4 data recovery Scn1a +/- mice. Iphone 5 data recovery Beginning at P18, F1. Top 5 data recovery software Scn1a +/- mice were fed either chow containing clobazam (320 mg per kg of chow; estimated dosage 40 mg/kg/day) or control chow, and survival was monitored to 30 days of age. Iphone 5 data recovery software This dose of clobazam, selected to achieve plasma concentrations within the human therapeutic range, was not sufficient to provide protection against survival. Raid 5 data recovery We used the hyperthermia-induced seizure assay to perform an acute dose-response study. Iphone 5 data recovery without backup Between P14–16, F1. Iphone 5 data recovery software free Scn1a +/- mice received an intraperitoneal injection of vehicle or clobazam (0.3, 1, or 30 mg/kg) prior to the induction of hyperthermia. Nexus 5 data recovery Clobazam administration provided dose-dependent protection against hyperthermia-induced seizures ( Fig 4). Raid 5 data recovery software Clobazam dosed at 30 mg/kg provided complete protection against hyperthermia-induced seizures, while 83% of vehicle controls exper ienced GTC seizures with an average threshold temperature of 42.3°C (p < 0.0006). Iphone 6 data recovery Clobazam doses of 0.3 and 1 mg/kg significantly increased GTC seizure thresholds to 43.0°C (p < 0.0022) and 42.9°C (p < 0.0079), respectively. Iphone 6 data recovery software To ensure plasma concentrations of clobazam reflected the human therapeutic range (0.1–0.4 μg/mL), plasma samples were assayed by HPLC. Iphone 6 data recovery free The commonly cited clobazam rodent dose of 30 mg/kg [ 20– 22] resulted in an average plasma concentration that was 15 times higher (5.84 ug/mL) than the human therapeutic range, while the 1 mg/kg dose resulted in an average plasma concentration (0.16 ug/mL) within the range. Iphone 6 data recovery mac Plasma concentrations were below the detection limit of the HPLC assay following the 0.3 mg/kg dose. Raid 6 data recovery Candidate gene and expression analysis, along with pharmacological testing, identified Gabra2 as a putative modifier gene that influences survival in the Scn1a +/- Dravet mouse model. Iphone 6 data recovery software free Considering the significance of GABAergic pathways in relation to epilepsy, strain-specific expression of Gabra2 may differentially influence excitatory/inhibitory balance. 7 data recovery review Several GWAS studies have demonstrated that human GABRA2 genetic variants are associated with alcohol dependence [ 23– 27]. 7 data recovery suite key However, to date, no GABRA2 genetic variants have been associated with human epilepsy. 7 data recovery suite crack Altered GABRA2 transcript and protein expression has observed in a number of rodent seizure models and patients with focal cortical dysplasia, tuberous sclerosis and temporal lobe epilepsy, suggesting a potential link between GABRA2 expression and epilepsy [ 28– 32]. 7 data recovery keygen There is evidence for variation in human cortical GABRA2 expression that is associated with a trans eQTL on chromosome Xp11.4 [ 33]. 7 data recovery key Our mouse RNA-Seq and expression analysis demonstrated that low Gabra2 expression associated with the B6 allele and correlated with reduced survival in the Scn1a +/- model. 7 data recovery serial key Previous work has shown that in some mouse strains, Gabra2 expression is strongly modulated by a cis eQTL that overlaps with Dsm1 and a trans eQTL on chromosome 10 [ 34]. 7 data recovery registration code The difference in Gabra2 expression was maintained when we examined transcript and protein levels in brain tissues from the ISC-D line, which contains 129 alleles only in the minimal interval and is B6 elsewhere in the genome. 7 data recovery suite registration code From this, we infer that Gabra2 expression difference arises from a local eQTL effect within the Dsm1 interval. Data recovery windows 8 This is supported by a recent study that demonstrated differential allele expression of Gabra2 transcript in B6/129SF1 mice, with the 129 allele expressed at 2–3 times the level of the B6 allele [ 35]. Windows 8 data recovery Future work will focus on identifying the responsible regulatory variants within the Gabra2 genomic region. Windows 8 data recovery software The failure of line ISC-D to reduce seizure frequency in F1. Data recovery 94fbr Scn1a +/- mice was not surprising. Easeus data recovery 9 serial As we report here, and have observed in our previous studies, high seizure counts do not correlate with premature death in F1. Icare data recovery 94fbr Scn1a +/- mice. Easeus data recovery 94fbr Therefore, we could exclude the survival improvement of the ISC-D line to be a result of altered seizure frequency. 7 data recovery 94fbr GABAergic neurons are found in several brain regions involved in the control of respiration and cardiac activity, and dysfunction in either pathway is believed to be primarily responsible for SUDEP [ 36– 39]. Tally erp 9 data recovery Mice with a conditional Scn1a +/- deletion in forebrain GABAergic interneurons experience SUDEP and cardiac dysfunctions similar to the Dravet Scn1a +/- mouse model [ 40]. Tally erp 9 data recovery software When Scn1a +/- was conditionally deleted in the heart, the phenotype was not observed, demonstrating that SUDEP observed in Scn1a +/- mice originates from the central nervous system [ 40]. Data recovery top 10 Increased expression of GABRA2 may exert a protective effect by altering autonomic control of cardiac and/or respiratory function and may explain the improved survival observed with 129/129 homozygosity in ISC-D. Top 10 data recovery software Additionally, GABA A receptor activity plays a pivotal role in neuronal development and altered levels of Gabra2 during development may alter network excitability [ 41]. Top 10 data recovery Interestingly, clobazam, which has preferential affinity for the GABRA2 subunit, offers some therapeutic benefit in Dravet syndrome patients [ 17– 19, 42, 43]. Top 10 data recovery software free download In our studies, threshold temperature for hyperthermia-induced seizures was elevated in Scn1a +/- mice treated with supratherapeutic levels of clobazam, while lower doses did not provide significant protection from hyperthermia-induced seizures or premature death. Top 10 data recovery software 2014 The 30mg/kg clobazam dose provided complete protection against hyperthermia-induced seizures, but resulted in plasma levels far above the human therapeutic range, highlighting the necessity of determining drug concentrations in similar studies. Top 10 data recovery software free Frequently, Dravet syndrome patients are prescribed stiripentol as an add-on treatment to clobazam. Top 10 data recovery tools In a randomized placebo-controlled study in children with Dravet syndrome, 71% of patients treated with a combination of clobazam and stiripentol experienced a reduction in seizure frequency [ 44]. Database administrator jobs Although the mechanism of action for the anticonvulsant benefit of stiripentol is yet to be determined, stiripentol has been identified as an inhibitor of CYP450s, specifically CYP3A4 and more potently CYP2C19, both important for the metabolism of clobazam. Database administrator salary In vivo, a significant increase in the plasma concentrations of both clobazam and its active metabolite N-desmethylclobazam has been shown with concomitant stiripentol treatment [ 45], suggesting that elevated levels of clobazam may account for some of the therapeutic benefit. Database administrator job description Identification of Gabra2 as a putative modifier gene at the Dsm1 locus furthers our understanding of the genetic basis of Dravet syndrome. How to become a database administrator This provides information that may help predict the risk of SUDEP and the clinical course of epilepsy due to a sodium channel mutation. Database management system Furthermore, modifier genes may suggest new targets for the improved treatment of epilepsy. Database management systems We generated five ISC lines carrying 129-derived chromosome 5 segments on a B6 background. Relational database management system [129 x B6]F1 progeny were continually backcrossed to B6 to generate congenic lines. Best database software Genotyping for chromosome 5 markers was performed at each generation and animals retaining 129 alleles in the Dsm1 interval were propagated. Free database software Whole genome and selective genotyping was performed at generations N2–N7 to select breeders with the lowest percentage of 129 alleles in the rest of the genome. Normalization in database All ISC lines were backcrossed to B6 for ≥N6 generations prior to experiments. Database administrator salary nyc 13. Database developer salary Lieberman R, Kranzler HR, Joshi P, Shin D-G, Covault J (2015) GABRA2 Alcohol Dependence Risk Allele is Associated with Reduced Expression of Chromosome 4p12 GABAA Subunit Genes in Human Neural Cultures. Fundamentals of database systems Alcoholism: Clinical and Experimental Research 39: 1654–1664. Google database [ PMC free article] [ PubMed] 28. Google hacking database Bethmann K, Fritschy JM, Brandt C, Loscher W (2008) Antiepileptic drug resistant rats differ from drug responsive rats in GABA A receptor subunit expression in a model of temporal lobe epilepsy. Gif database Neurobiol Dis 31: 169–187. Graph database doi: 10.1016/j.nbd.2008.01.005 [ PubMed]