Characterisation of Cdkl5 transcript isoforms in rat

Hector, Ralph D.; Dando, Owen; Kind, Peter C.; Bailey, Mark E.S.; Cobb, Stuart

doi:10.1016/j.gene.2016.12.001

Cited by 12 publications

(13 citation statements)

References 16 publications

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“…In mice, CDKL5 is expressed throughout the brain starting at late embryonic stages and through adulthood, indicating a role in postnatal development and function in the nervous system (Chen et al, 2010;Hector et al, 2016Hector et al, , 2017. CDKL5 mRNA is highly enriched in foetal and adult human brains (Hector et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Chemical genetic identification of CDKL 5 substrates reveals its role in neuronal microtubule dynamics

et al. 2018

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Loss‐of‐function mutations in CDKL5 kinase cause severe neurodevelopmental delay and early‐onset seizures. Identification of CDKL5 substrates is key to understanding its function. Using chemical genetics, we found that CDKL5 phosphorylates three microtubule‐associated proteins: MAP1S, EB2 and ARHGEF2, and determined the phosphorylation sites. Substrate phosphorylations are greatly reduced in CDKL5 knockout mice, verifying these as physiological substrates. In CDKL5 knockout mouse neurons, dendritic microtubules have longer EB3‐labelled plus‐end growth duration and these altered dynamics are rescued by reduction of MAP1S levels through shRNA expression, indicating that CDKL5 regulates microtubule dynamics via phosphorylation of MAP1S. We show that phosphorylation by CDKL5 is required for MAP1S dissociation from microtubules. Additionally, anterograde cargo trafficking is compromised in CDKL5 knockout mouse dendrites. Finally, EB2 phosphorylation is reduced in patient‐derived human neurons. Our results reveal a novel activity‐dependent molecular pathway in dendritic microtubule regulation and suggest a pathological mechanism which may contribute to CDKL5 deficiency disorder.

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Section: Introductionmentioning

confidence: 99%

Chemical genetic identification of CDKL 5 substrates reveals its role in neuronal microtubule dynamics

et al. 2018

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“…hCDKL5_1-4 are ubiquitously expressed, whilst hCDKL5_5 is expressed only in the adult testes and foetal brain (Hector et al, 2016). The majority of the CDKL5 coding region is orthologous and well-conserved between human, rat and mouse (Hector et al, 2016(Hector et al, , 2017a.…”

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confidence: 99%

Gene replacement ameliorates deficits in mouse and human models of cyclin-dependent kinase-like 5 disorder

Gao

Irvine

Eleftheriadou

et al. 2020

Brain

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Cyclin-dependent kinase-like 5 disorder is a severe neurodevelopmental disorder caused by mutations in the X-linked cyclin-dependent kinase-like 5 (CDKL5) gene. It predominantly affects females who typically present with severe early epileptic encephalopathy, global developmental delay, motor dysfunction, autistic features and sleep disturbances. To develop a gene replacement therapy, we initially characterized the human CDKL5 transcript isoforms expressed in the brain, neuroblastoma cell lines, primary astrocytes and embryonic stem cell-derived cortical interneurons. We found that the isoform 1 and to a lesser extent the isoform 2 were expressed in human brain, and both neuronal and glial cell types. These isoforms were subsequently cloned into recombinant adeno-associated viral (AAV) vector genome and high-titre viral vectors were produced. Intrajugular delivery of green fluorescence protein via AAV vector serotype PHP.B in adult wild-type male mice transduced neurons and astrocytes throughout the brain more efficiently than serotype 9. Cdkl5 knockout male mice treated with isoform 1 via intrajugular injection at age 28–30 days exhibited significant behavioural improvements compared to green fluorescence protein-treated controls (1012 vg per animal, n = 10 per group) with PHP.B vectors. Brain expression of the isoform 1 transgene was more abundant in hindbrain than forebrain and midbrain. Transgene brain expression was sporadic at the cellular level and most prominent in hippocampal neurons and cerebellar Purkinje cells. Correction of postsynaptic density protein 95 cerebellar misexpression, a major fine cerebellar structural abnormality in Cdkl5 knockout mice, was found in regions of high transgene expression within the cerebellum. AAV vector serotype DJ efficiently transduced CDKL5-mutant human induced pluripotent stem cell-derived neural progenitors, which were subsequently differentiated into mature neurons. When treating CDKL5-mutant neurons, isoform 1 expression led to an increased density of synaptic puncta, while isoform 2 ameliorated the calcium signalling defect compared to green fluorescence protein control, implying distinct functions of these isoforms in neurons. This study provides the first evidence that gene therapy mediated by AAV vectors can be used for treating CDKL5 disorder.

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“…CDKL5 is mostly expressed in neurons and its expression in glial cells is extremely low (Rusconi et al, 2008;Chen et al, 2010). Multiple splicing isoforms of CDKL5 have been reported in different species, and some of them are predominantly expressed in the brain (Chen et al, 2010;Fichou et al, 2011;Williamson et al, 2012;Hector et al, 2016;Hector et al, 2017). The functional significance of these spicing events is not clear.…”

Section: Developmental Neurobiologymentioning

confidence: 99%

Molecular and Synaptic Bases of CDKL5 Disorder

Yingguo

Xiong

2018

Developmental Neurobiology

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The X‐linked gene cyclin‐dependent kinase‐like 5 (CDKL5) encodes a serine/threonine kinase abundantly expressed in the brain. Mutations in CDKL5 have been associated with neurodevelopmental disorders characterized by early‐onset epileptic encephalopathy and severe intellectual disability, suggesting that CDKL5 plays important roles in brain development and function. Recent studies using cultured neurons, knockout mice, and human iPSC‐derived neurons have demonstrated that CDKL5 regulates axon outgrowth, dendritic morphogenesis, and synapse formation. The role of CDKL5 in maintaining synaptic function in the mature brain has also begun to emerge. Moreover, mouse models that are deficient for CDKL5 recapitulate some of the key clinical phenotypes in human patients. Here we review these findings related to the function of CDKL5 in the brain and discuss the underlying molecular and cellular mechanisms.

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Characterisation of Cdkl5 transcript isoforms in rat

Cited by 12 publications

References 16 publications

Chemical genetic identification of CDKL 5 substrates reveals its role in neuronal microtubule dynamics

Chemical genetic identification of CDKL 5 substrates reveals its role in neuronal microtubule dynamics

Gene replacement ameliorates deficits in mouse and human models of cyclin-dependent kinase-like 5 disorder

Molecular and Synaptic Bases of CDKL5 Disorder

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