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Calsequestrins New Calcium Store Markers of Adult Zebrafish Cerebellum and Optic Tectum.
Frontiers in Neuroanatomy ( IF 2.9 ) Pub Date : 2020-03-18 , DOI: 10.3389/fnana.2020.00015
Sandra Furlan 1 , Marina Campione 1 , Marta Murgia 2, 3 , Simone Mosole 4 , Francesco Argenton 5 , Pompeo Volpe 2 , Alessandra Nori 2
Affiliation  

Calcium stores in neurons are heterogeneous in compartmentalization and molecular composition. Danio rerio (zebrafish) is an animal model with a simply folded cerebellum similar in cellular organization to that of mammals. The aim of the study was to identify new endoplasmic reticulum (ER) calcium store markers in zebrafish adult brain with emphasis on cerebellum and optic tectum. By quantitative polymerase chain reaction, we found three RNA transcripts coding for the intra-ER calcium binding protein calsequestrin: casq1a, casq1b, and casq2. In brain homogenates, two isoforms were detected by mass spectrometry and western blotting. Fractionation experiments of whole brain revealed that Casq1a and Casq2 were enriched in a heavy fraction containing ER microsomes and synaptic membranes. By in situ hybridization, we found the heterogeneous expression of casq1a and casq2 mRNA to be compatible with the cellular localization of calsequestrins investigated by immunofluorescence. Casq1 was expressed in neurogenic differentiation 1 expressing the granule cells of the cerebellum and the periventricular zone of the optic tectum. Casq2 was concentrated in parvalbumin expressing Purkinje cells. At a subcellular level, Casq1 was restricted to granular cell bodies, and Casq2 was localized in cell bodies, dendrites, and axons. Data are discussed in relation to the differential cellular and subcellular distribution of other cerebellum calcium store markers and are evaluated with respect to the putative relevance of calsequestrins in the neuron-specific functional activity.



中文翻译:

Calsequestrins成年斑马鱼小脑和视神经直肠的钙存储新标记。

神经元中的钙存储在区室化和分子组成上是异质的。 里约热内卢斑马鱼(斑马鱼)是一种具有简单折叠的小脑的动物模型,其小脑组织在细胞结构上与哺乳动物相似。该研究的目的是在斑马鱼成年大脑中鉴定新的内质网(ER)钙存储标记,重点是小脑和视神经支架。通过定量聚合酶链反应,我们发现了三个编码ER内钙结合蛋白calsequestrin的RNA转录物:卡斯卡1acasq1bcasq2。在脑匀浆中,通过质谱和蛋白质印迹检测到两个同工型。全脑的分级实验表明,Casq1a和Casq2富含含有ER微粒体和突触膜的重组分。通过原位 杂交,我们发现 卡斯卡1acasq2通过免疫荧光研究与钙锚蛋白的细胞定位相容的mRNA。Casq1在神经源性分化1中表达,该分化表达小脑的颗粒细胞和视神经外膜的脑室周围区域。Casq2浓缩在表达小白蛋白的Purkinje细胞中。在亚细胞水平,Casq1仅限于颗粒状细胞体,而Casq2则位于细胞体,树突和轴突中。讨论了与其他小脑钙存储标记物在细胞和亚细胞中的分布差异有关的数据,并评估了钙螯合蛋白在神经元特异性功能活性中的假定相关性。

更新日期:2020-03-18
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