Mosaic and non-mosaic protocadherin 19 mutation leads to neuronal hyperexcitability in zebrafish

Robens, Barbara K.; Yang, Xinzhu; McGraw, Christopher M.; Turner, Laura H.; Robens, Carsten; Thyme, Summer B.; Rotenberg, Alexander; Poduri, Annapurna

doi:10.1016/j.nbd.2022.105738

Cited by 7 publications

(4 citation statements)

References 44 publications

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“…Pcdh19 crispants also showed an increased response to flashing light while ube3a inactivation only had a minor impact on light-induced seizures. Interestingly, pcdh19 has been previously associated with neuronal hyperactivity in zebrafish [ 67 ], but, to our knowledge, this study demonstrated for the first time its association with seizure-like behavior.…”

Section: Discussionmentioning

confidence: 61%

A Zebrafish-Based Platform for High-Throughput Epilepsy Modeling and Drug Screening in F0

Locubiche,

Ordóñez,

Abad

et al. 2024

IJMS

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The zebrafish model has emerged as a reference tool for phenotypic drug screening. An increasing number of molecules have been brought from bench to bedside thanks to zebrafish-based assays over the last decade. The high homology between the zebrafish and the human genomes facilitates the generation of zebrafish lines carrying loss-of-function mutations in disease-relevant genes; nonetheless, even using this alternative model, the establishment of isogenic mutant lines requires a long generation time and an elevated number of animals. In this study, we developed a zebrafish-based high-throughput platform for the generation of F0 knock-out (KO) models and the screening of neuroactive compounds. We show that the simultaneous inactivation of a reporter gene (tyrosinase) and a second gene of interest allows the phenotypic selection of F0 somatic mutants (crispants) carrying the highest rates of mutations in both loci. As a proof of principle, we targeted genes associated with neurodevelopmental disorders and we efficiently generated de facto F0 mutants in seven genes involved in childhood epilepsy. We employed a high-throughput multiparametric behavioral analysis to characterize the response of these KO models to an epileptogenic stimulus, making it possible to employ kinematic parameters to identify seizure-like events. The combination of these co-injection, screening and phenotyping methods allowed us to generate crispants recapitulating epilepsy features and to test the efficacy of compounds already during the first days post fertilization. Since the strategy can be applied to a wide range of indications, this study paves the ground for high-throughput drug discovery and promotes the use of zebrafish in personalized medicine and neurotoxicity assessment.

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Section: Discussionmentioning

confidence: 61%

A Zebrafish-Based Platform for High-Throughput Epilepsy Modeling and Drug Screening in F0

Locubiche,

Ordóñez,

Abad

et al. 2024

IJMS

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“…Variants can cause changes in protein structure and function in a variety of ways, including interference of homophilic adhesion, impairment of interaction with other calcium adhesion proteins, and a decrease in protein folding function and stability ( 23 ). Recent experiments on animal models illustrated that the PCDH19 variant impaired the mossy fiber synapse development in a mouse model, and led to neuronal hyperexcitability in both mosaic and non-mosaic PCDH19 mutation zebrafish models ( 24 , 25 ). The pathophysiology of PCDH19 -related epilepsy has also been reviewed recently with four main theories: gamma-aminobutyric acid type A receptor [GABAA(R)] dysregulation, cellular interference, blood-brain barrier dysfunction, and the aldo-keto reductase 1C family 1-3 (AKR1C1-3) gene product shortage ( 26 ).…”

Section: Discussionmentioning

confidence: 99%

Genetic variants and phenotype analysis in a five-generation Chinese pedigree with PCDH19 female-limited epilepsy

Zhou

Ouyang²,

Ji³

et al. 2023

Front. Neurol.

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ObjectiveAlbeit the gene of PCDH19-FE was ascertained, the correlation of gene mutation, PCDH19 protein structure, and phenotype heterogeneity remained obscure. This study aimed to report a five-generation pedigree of seven female patients of PCDH19-FE and tried to explore whether two variants were correlated with PCDH19 protein structure and function alteration, and PCDH19-FE phenotype.MethodsWe analyzed the clinical data and genetic variants of a PCDH19-FE pedigree, to explore the phenotype heterogeneity of PCDH19-FE and underlying mechanisms. In addition to the clinical information of family members, next-generation sequencing was adopted to detect the variant sites of probands with validation by sanger sequencing. And the sanger sequencing was conducted in other patients in this pedigree. The biological conservation analysis and population polymorphism analysis of variants were also performed subsequently. The structure alteration of mutated PCDH19 protein was predicted by AlphaFold2.ResultsBased on a five-generation pedigree of PCDH19-FE, missense variants of c.695A>G and c.2760T>A in the PCDH19 gene were found in the heterozygous proband (V:1), which resulted in the change of amino acid 232 from Asn to Ser (p.Asn232Ser) and amino acid 920 from Asp to Glu (p.Asp920Glu) influencing PCDH19 function. The other six females in the pedigree (II:6, II:8, IV:3, IV:4, IV:5, IV:11) exhibited different clinical phenotypes but shared the same variant. Two males with the same variant have no clinical manifestations (III:3, III:10). The biological conservation analysis and population polymorphism analysis demonstrated the highly conservative characteristics of these two variants. AlphaFold2 predicted that the variant, p.Asp920Glu, led to the disappearance of the hydrogen bond between Asp at position 920 and His at position 919. Furthermore, the hydrogen bond between Asp920 and His919 also disappeared when the Asn amino acid mutated to Ser at position 232.ConclusionA strong genotype-phenotype heterogeneity was observed among female patients with the same genotype in our PCDH19-FE pedigree. And two missense variants, c.695A > G and c.2760T>A in the PCDH19 gene, have been identified in our pedigree. The c.2760T>A variant was a novel variant site probably related to the PCDH19-FE.

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“…At present, the variable expression of epilepsy genes is known, and some cases could be carriers and show only EEG abnormalities. Mosaic carriers may explain this phenomenon ( Robens et al, 2022 ). Here, we reported another DHDDS mutation which presented with autosomal dominant inheritance pattern and lead to intractable epilepsy and/or myoclonus.…”

Section: Discussionmentioning

confidence: 99%

Case report: Identification of a recurrent pathogenic DHDDS mutation in Chinese family with epilepsy, intellectual disability and myoclonus

Dong,

Zhang,

Sheng

et al. 2023

Front. Genet.

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Background: Heterozygous mutations in the dehydrodolichol diphosphate synthase (DHDDS) gene are one of the causes generating developmental and epileptic encephalopathies. So far, only eleven mutations in the DHDDS gene have been identified. The mutation spectrum of the DHDDS gene in the Chinese population remains unclear.Methods: In this study, we enrolled a Chinese family with myoclonus and/or epilepsy and intellectual disability. The epilepsy and myoclonic tremor were improved after deep brain stimulation (DBS) of the subthalamic nucleus (STN) treatment. Whole exome sequencing and Sanger sequencing were employed to explore the genetic variations of the family.Results: Subsequent to data filtering, we identified a recurrent pathogenic mutation (NM_001243564.1, c.113G>A/p.R38H) in the DHDDS gene in the proband. Sanger sequencing further validated that the presence of the mutation in his affected mother but absent in the health family members. Further bioinformatics analysis revealed that this mutation (p.R38H), located in an evolutionarily conserved region of DHDDS, was predicted to be deleterious.Discussion: In this report, we present the first case of intractable epilepsy and/or myoclonus caused by p.R38H mutation of the DHDDS gene in the Chinese population. Furthermore, this study represents the third report of autosomal dominant familial inheritance of DHDDS mutation worldwide.

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Mosaic and non-mosaic protocadherin 19 mutation leads to neuronal hyperexcitability in zebrafish

Cited by 7 publications

References 44 publications

A Zebrafish-Based Platform for High-Throughput Epilepsy Modeling and Drug Screening in F0

A Zebrafish-Based Platform for High-Throughput Epilepsy Modeling and Drug Screening in F0

Genetic variants and phenotype analysis in a five-generation Chinese pedigree with PCDH19 female-limited epilepsy

Case report: Identification of a recurrent pathogenic DHDDS mutation in Chinese family with epilepsy, intellectual disability and myoclonus

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