Single Amino Acid Changes in the Ryanodine Receptor in the Human Population Have Effects In Vivo on Caenorhabditis elegans Neuro-Muscular Function
Single Amino Acid Changes in the Ryanodine Receptor in the Human Population Have Effects In Vivo on Caenorhabditis elegans Neuro-Muscular Function
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DOI：
10.3389/fgene.2020.00037
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发表时间：
2020-02-26
影响因子：
3.7
通讯作者：
Hope, Ian A.
中科院分区：
文献类型：
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作者：
Graham, Brittany;Shaw, Marie-Anne;Hope, Ian A.
兰尼碱受体介导神经和肌肉细胞兴奋时细胞内钙离子释放。Ryanodine受体错义变体引起许多肌肉病理学，例如恶性高热，并且与包括阿尔茨海默病在内的各种神经病理学有关。我们的特点是在体内的兰尼碱受体变异体的后果。通过基因组编辑产生了八种秀丽隐杆线虫菌株，其ryanodine受体修饰相当于人类肌病RYR1变体。在人类中，这些变异是罕见的，并赋予敏感性吸入麻醉剂氟烷时杂合子。在纯合子和杂合子C.优雅的仔细分析发现，由于不同的单个氨基酸残基的变化，即使在没有外部触发剂的情况下，不同的微妙的运动缺陷。不同的前和突触后的后果的变体，其特征在于通过胆碱能药物的反应。表型的范围反映了不同细胞类型中随着年龄的增长，兰尼碱受体调节输入的复杂性以及钙离子通道开放特性的关键性。具有这些单个氨基酸残基变化的兰尼碱受体仍然作为钙离子通道起作用，但是具有改变的性质，这可能对这些变体的人类携带者具有微妙的后果。钙离子信号微妙改变的长期后果可能是累积的，并且可能集中在较小的神经细胞中，而不是更健壮的肌肉细胞中。重要的是评估体内表型，以正确理解整个生物体的后果。
The ryanodine receptor mediates intracellular calcium ion release with excitation of nerve and muscle cells. Ryanodine receptor missense variants cause a number of myopathologies, such as malignant hyperthermia, and have been linked with various neuropathologies, including Alzheimer's disease. We characterized the consequences of ryanodine receptor variants in vivo. Eight Caenorhabditis elegans strains, with ryanodine receptor modifications equivalent to human myopathic RYR1 variants, were generated by genome editing. In humans, these variants are rare and confer sensitivity to the inhalational anaesthetic halothane when heterozygous. Increased sensitivity to halothane was found in both homozygous and heterozygous C. elegans. Close analysis revealed distinct subtle locomotion defects, due to the different single amino acid residue changes, even in the absence of the external triggering agent. Distinct pre- and postsynaptic consequences of the variants were characterized through the responses to cholinergic pharmacological agents. The range of phenotypes reflects the complexity of the regulatory inputs to the ryanodine receptor and the criticality of the calcium ion channel opening properties, in different cell types and with age. Ryanodine receptors with these single amino acid residue changes still function as calcium ion channels, but with altered properties which are likely to have subtle consequences for human carriers of such variants. The long-term consequences of subtly altered calcium ion signalling could be cumulative and may be focussed in the smaller nerve cells rather than the more robust muscle cells. It was important to assess phenotypes in vivo to properly appreciate consequences for a whole organism.