Heat induced temperature dysregulation and seizures in Dravet Syndrome/GEFS+ Gabrg2+/Q390X mice

Warner, Timothy A.; Liu, Zhong; Macdonald, Robert L.; Kang, Jing‐Qiong

doi:10.1016/j.eplepsyres.2017.04.020

Cited by 22 publications

(26 citation statements)

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“…We avoided longer heat exposure, because sustained high‐temperature exposure may cause cellular injury, and even cell death. The temperature induction protocol was modified based on previously described studies in both Gabrg2 +/Q390X and Scn1a +/‐ KO mice 20,29 (Figure 3A). Heating stopped once the mouse's core temperature reached 42.5°C, but this temperature was maintained for 30 minutes before tissue harvest (Figure 3B).…”

Section: Resultsmentioning

confidence: 99%

Endoplasmic reticulum stress increases inflammatory cytokines in an epilepsy mouse model Gabrg2^+/Q390X knockin: A link between genetic and acquired epilepsy?

et al. 2020

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Objective: Neuroinflammation is a major theme in epilepsy, which has been characterized in acquired epilepsy but is poorly understood in genetic epilepsy. γ-Aminobutyric acid type A receptor subunit gene mutations are significant causes of epilepsy, and we have studied the pathophysiology directly resulting from defective receptor channels. Here, we determined the proinflammatory factors in a genetic mouse model, the Gabrg2 +/Q390X knockin (KI). We have identified increased cytokines in multiple brain regions of the KI mouse throughout different developmental stages and propose that accumulation of the trafficking-deficient mutant protein may increase neuroinflammation, which would be a novel mechanism for genetic epilepsy. Methods: We used enzyme-linked immunosorbent assay, immunoprecipitation, nuclei purification, immunoblot, immunohistochemistry, and confocal microscopy to characterize increased neuroinflammation and its potential causes in a Gabrg2 +/Q390X KI mouse and a Gabrg2 +/knockout (KO) mouse, each associated with a different epilepsy syndrome with different severities. Results: We found that proinflammatory cytokines such as tumor necrosis factor alpha, interleukin 1-beta (IL-1β), and IL-6 were increased in the KI mice but not in the KO mice. A major underlying basis for the discrepancy in cytokine expression between the two mouse models is likely chronic mutant protein accumulation and endoplasmic reticulum (ER) stress. The presence of mutant protein dampened cytokine induction upon further cellular stimulation or external stress such as elevated temperature. Pharmacological induction of ER stress upregulated cytokine expression in the wild-type and KO but not in the KI mice. The increased cytokine expression was independent of seizure occurrence, because it was upregulated in both mice and cultured neurons. Significance: Together, these data demonstrate a novel pathophysiology for genetic epilepsy, increased neuroinflammation, which is a common mechanism for acquired 2302 | SHEN Et al.

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Section: Resultsmentioning

confidence: 99%

Endoplasmic reticulum stress increases inflammatory cytokines in an epilepsy mouse model Gabrg2^+/Q390X knockin: A link between genetic and acquired epilepsy?

et al. 2020

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“…For both slc7a5 and syngap1b, we did not observe any strong mutational effects in non-PTZ treated larvae. For future studies, we will consider incorporating additional perturbations during motion tracking with known associations to enhanced behaviors and seizure phenotypes, such as heat (Warner et al, 2017;Peron et al, 2018) and startle (Tegelenbosch et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform

Colón-Rodríguez

Uribe-Salazar

Weyenberg

et al. 2020

Front. Cell Dev. Biol.

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In recent years, zebrafish have become commonly used as a model for studying human traits and disorders. Their small size, high fecundity, and rapid development allow for more high-throughput experiments compared to other vertebrate models. Given that zebrafish share >70% gene homologs with humans and their genomes can be readily edited using highly efficient CRISPR methods, we are now able to rapidly generate mutations impacting practically any gene of interest. Unfortunately, our ability to phenotype mutant larvae has not kept pace. To address this challenge, we have developed a protocol that obtains multiple phenotypic measurements from individual zebrafish larvae in an automated and parallel fashion, including morphological features (i.e., body length, eye area, and head size) and movement/behavior. By assaying wild-type zebrafish in a variety of conditions, we determined optimal parameters that avoid significant developmental defects or physical damage; these include morphological imaging of larvae at two time points [3 days post fertilization (dpf) and 5 dpf] coupled with motion tracking of behavior at 5 dpf. As a proof-of-principle, we tested our approach on two novel CRISPR-generated mutant zebrafish lines carrying predicted null-alleles of syngap1b and slc7a5, orthologs to two human genes implicated in autism-spectrum disorder, intellectual disability, and epilepsy. Using our optimized high-throughput phenotyping protocol, we recapitulated previously published results from mouse and zebrafish models of these candidate genes. In summary, we describe a rapid parallel pipeline to characterize morphological and behavioral features of individual larvae in a robust and consistent fashion, thereby improving our ability to better identify genes important in human traits and disorders.

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Section: Discussionmentioning

confidence: 99%

Assessment of autism zebrafish mutant models using a high-throughput larval phenotyping platform

Colón-Rodríguez

Uribe-Salazar²,

Weyenberg³

et al. 2020

Preprint

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In recent years zebrafish have become commonly used as a model for studying human traits and disorders. Their small size, high fecundity, and rapid development allow for more high-throughput experiments compared to other vertebrate models. Given that zebrafish share >70% gene homologs with humans and their genomes can be readily edited using highly efficient CRISPR methods, we are now able to rapidly generate mutations impacting practically any gene of interest. Unfortunately, our ability to phenotype mutant larvae has not kept pace. To address this challenge, we have developed a protocol that obtains multiple phenotypic measurements from individual zebrafish larvae in an automated and parallel fashion, including morphological features (i.e., body length, eye area, and head size) and movement/behavior. By assaying wild-type zebrafish in a variety of conditions, we determined optimal parameters that avoid significant developmental defects or physical damage; these include morphological imaging of larvae at two time points (3 days post fertilization (dpf) and 5 dpf) coupled with motion tracking of behavior at 5 dpf. As a proof-of-principle, we tested our approach on two novel CRISPR-generated mutant zebrafish lines carrying predicted null-alleles of syngap1b and slc7a5, orthologs to two human genes implicated in autism-spectrum disorder, intellectual disability, and epilepsy. Using our optimized high-throughput phenotyping protocol, we recapitulated previously published results from mouse and zebrafish models of these candidate genes. In summary, we describe a rapid parallel pipeline to characterize morphological and behavioral features of individual larvae in a robust and consistent fashion, thereby improving our ability to better identify genes important in human traits and disorders.AUTHOR SUMMARYZebrafish (Danio rerio) are a well-established model organism for the study of neurodevelopmental disorders. Due to their small size, fast reproduction, and genetic homology with humans, zebrafish have been widely used for characterizing and screening candidate genes for many disorders, including autism-spectrum disorder, intellectual disability, and epilepsy. Although several studies have described the use of high-throughput morphological and behavioral assays, few combine multiple assays in a single zebrafish larva. Here, we optimized a platform to characterize morphometric features at two developmental time points in addition to behavioral traits of zebrafish larvae. We then used this approach to characterize two autism candidate genes (SYNGAP1 and SLC7A5) in two CRISPR-generated zebrafish null mutant models we developed in house. These data recapitulate previously published results related to enhanced seizure activity, while identifying additional defects not previously reported. We propose that our phenotyping platform represents a feasible method for maximizing the use of single zebrafish larvae in the characterization of additional mutants relevant to neurodevelopmental disorders.

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Heat induced temperature dysregulation and seizures in Dravet Syndrome/GEFS+ Gabrg2+/Q390X mice

Cited by 22 publications

References 44 publications

Endoplasmic reticulum stress increases inflammatory cytokines in an epilepsy mouse model Gabrg2^+/Q390X knockin: A link between genetic and acquired epilepsy?

Endoplasmic reticulum stress increases inflammatory cytokines in an epilepsy mouse model Gabrg2^+/Q390X knockin: A link between genetic and acquired epilepsy?

Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform

Assessment of autism zebrafish mutant models using a high-throughput larval phenotyping platform

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Heat induced temperature dysregulation and seizures in Dravet Syndrome/GEFS+ Gabrg2+/Q390X mice

Cited by 22 publications

References 44 publications

Endoplasmic reticulum stress increases inflammatory cytokines in an epilepsy mouse model Gabrg2+/Q390X knockin: A link between genetic and acquired epilepsy?

Endoplasmic reticulum stress increases inflammatory cytokines in an epilepsy mouse model Gabrg2+/Q390X knockin: A link between genetic and acquired epilepsy?

Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform

Assessment of autism zebrafish mutant models using a high-throughput larval phenotyping platform

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Product

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Endoplasmic reticulum stress increases inflammatory cytokines in an epilepsy mouse model Gabrg2^+/Q390X knockin: A link between genetic and acquired epilepsy?

Endoplasmic reticulum stress increases inflammatory cytokines in an epilepsy mouse model Gabrg2^+/Q390X knockin: A link between genetic and acquired epilepsy?