Mutations in the cyclin-dependent kinase-like 5 gene(CDKL5)cause a severe neurodevelopmental disorder,yet the impact of truncating mutations remains unclear.Here,we introduce the Cdkl5^(492stop) mouse model,mimicking ...Mutations in the cyclin-dependent kinase-like 5 gene(CDKL5)cause a severe neurodevelopmental disorder,yet the impact of truncating mutations remains unclear.Here,we introduce the Cdkl5^(492stop) mouse model,mimicking C-terminal truncating mutations in patients.492stop/Y mice exhibit altered dendritic spine morphology and spontaneous seizure-like behaviors,alongside other behavioral deficits.After creating cell lines with various Cdkl5 truncating mutations,we found that these mutations are regulated by the nonsense-mediated RNA decay pathway.Most truncating mutations result in CDKL5 protein loss,leading to multiple disease phenotypes,and offering new insights into the pathogenesis of CDKL5 disorder.展开更多
目的研究丝/苏氨酸激酶周期素依赖性激酶样-5(CDKL5)在胃癌组织中的表达,并分析其与胃癌患者的临床病理特征之间的关系。方法应用荧光实时定量PCR法和免疫印迹法分别检测45例胃癌及其癌旁组织中CDKL5 m RNA及蛋白的表达,然后用免疫荧光...目的研究丝/苏氨酸激酶周期素依赖性激酶样-5(CDKL5)在胃癌组织中的表达,并分析其与胃癌患者的临床病理特征之间的关系。方法应用荧光实时定量PCR法和免疫印迹法分别检测45例胃癌及其癌旁组织中CDKL5 m RNA及蛋白的表达,然后用免疫荧光方法检测CDKL5蛋白在AGS和MKN-45胃癌细胞株中的表达情况。结果 CDKL5 m RNA和蛋白在胃癌组织中的表达均明显高于其在相应的癌旁组织中的表达(P<0.05);CDKL5 m RNA的表达与胃癌患者的年龄(P=0.033)、血管浸润(P=0.007)、T分期(P=0.049)和TNM分期(P=0.041)有关;多重逐步回归分析结果显示,血管浸润(P=0.013)和TNM分期(P=0.024)是影响CDKL5m RNA表达的重要因素;胃癌组织和细胞株中CDKL5蛋白除了在相对分子质量为107×103条带处显影外,还检测到在相对分子质量为85×103处的显影条带。结论 CDKL5在胃癌组织和细胞中均呈高表达,其m RNA表达与胃癌患者的血管浸润和TNM分期有关;CDKL5蛋白分别在相对分子质量为107×103和85×103条带处显影,提示CDKL5在蛋白水平可能存在不同的修饰方式或者表达方式。展开更多
Developmental and epileptic encephalopathies are severe neurological conditions in clinical practice,among which loss-of-function mutations in brain-enriched serine-threonine kinase cyclin dependent kinase like-5(CDKL...Developmental and epileptic encephalopathies are severe neurological conditions in clinical practice,among which loss-of-function mutations in brain-enriched serine-threonine kinase cyclin dependent kinase like-5(CDKL5)exists as one of the most common types.It is unknown,therefore,how precisely CDKL5 mutations lead to neuronal hyper-excitation.A recent study that looked at the connection between voltage-gated calcium channel Cav2.3 and CDKL5 in an experimental context was published in Nature Communications.This study has revealed that Cav2.3,a physi-ological phosphorylation target of CDKL5,would show delayed inactivation and increased cholinergic stimulation in CDKL5 knock out conditions.This would in turn cause neuronal hyperexcitability and related enhanced seizure susceptibility.This work,in our opinion,provided fresh insight into the epileptic encephalopathies linked to CDKL5 and highlighted Cav2.3 as a possible target for it.展开更多
基金supported by the Innovation of Science and Technology 2030-Major Project"Platform of Nonhuman Primate Models"(2021ZD0200900)the Ministry of Science and Technology(2018YFA0801404)+1 种基金the National Natural Science Foundation of China(82021001)the Shanghai Municipal Science and Technology Major Project.
文摘Mutations in the cyclin-dependent kinase-like 5 gene(CDKL5)cause a severe neurodevelopmental disorder,yet the impact of truncating mutations remains unclear.Here,we introduce the Cdkl5^(492stop) mouse model,mimicking C-terminal truncating mutations in patients.492stop/Y mice exhibit altered dendritic spine morphology and spontaneous seizure-like behaviors,alongside other behavioral deficits.After creating cell lines with various Cdkl5 truncating mutations,we found that these mutations are regulated by the nonsense-mediated RNA decay pathway.Most truncating mutations result in CDKL5 protein loss,leading to multiple disease phenotypes,and offering new insights into the pathogenesis of CDKL5 disorder.
文摘目的研究丝/苏氨酸激酶周期素依赖性激酶样-5(CDKL5)在胃癌组织中的表达,并分析其与胃癌患者的临床病理特征之间的关系。方法应用荧光实时定量PCR法和免疫印迹法分别检测45例胃癌及其癌旁组织中CDKL5 m RNA及蛋白的表达,然后用免疫荧光方法检测CDKL5蛋白在AGS和MKN-45胃癌细胞株中的表达情况。结果 CDKL5 m RNA和蛋白在胃癌组织中的表达均明显高于其在相应的癌旁组织中的表达(P<0.05);CDKL5 m RNA的表达与胃癌患者的年龄(P=0.033)、血管浸润(P=0.007)、T分期(P=0.049)和TNM分期(P=0.041)有关;多重逐步回归分析结果显示,血管浸润(P=0.013)和TNM分期(P=0.024)是影响CDKL5m RNA表达的重要因素;胃癌组织和细胞株中CDKL5蛋白除了在相对分子质量为107×103条带处显影外,还检测到在相对分子质量为85×103处的显影条带。结论 CDKL5在胃癌组织和细胞中均呈高表达,其m RNA表达与胃癌患者的血管浸润和TNM分期有关;CDKL5蛋白分别在相对分子质量为107×103和85×103条带处显影,提示CDKL5在蛋白水平可能存在不同的修饰方式或者表达方式。
基金supported by the National Natural Science Foundation of China(82173796).Ava。
文摘Developmental and epileptic encephalopathies are severe neurological conditions in clinical practice,among which loss-of-function mutations in brain-enriched serine-threonine kinase cyclin dependent kinase like-5(CDKL5)exists as one of the most common types.It is unknown,therefore,how precisely CDKL5 mutations lead to neuronal hyper-excitation.A recent study that looked at the connection between voltage-gated calcium channel Cav2.3 and CDKL5 in an experimental context was published in Nature Communications.This study has revealed that Cav2.3,a physi-ological phosphorylation target of CDKL5,would show delayed inactivation and increased cholinergic stimulation in CDKL5 knock out conditions.This would in turn cause neuronal hyperexcitability and related enhanced seizure susceptibility.This work,in our opinion,provided fresh insight into the epileptic encephalopathies linked to CDKL5 and highlighted Cav2.3 as a possible target for it.
