Eskitis Institute for Drug Discovery, Griffith University , Brisbane, QLD, Australia.
NPJ Schizophr. 2016 Aug 17;2:16027. doi: 10.1038/npjschz.2016.27. eCollection 2016.
Reelin expression is reduced in various regions in the post-mortem brain of schizophrenia patients but the exact role of reelin function in the neurobiology of schizophrenia remains elusive. Absence of reelin in knockout mouse causes inverted lamination of the neocortex due to aberrant neuronal migration. The aim of this study was to utilize patient-derived olfactory neurosphere-derived (ONS) cells to investigate whether extracellular reelin alters cell motility in schizophrenia patient-derived cells. ONS cells from nine patients were compared with cells from nine matched healthy controls. Automated high-throughput imaging and analysis were used to track motility of individual living cells on reelin-coated surfaces produced from reelin secreted into the medium by HEK293FT cells transfected with the full-length reelin plasmid pCrl. Automated assays were used to quantify intracellular cytoskeleton composition, cell morphology, and focal adhesions. Expression of reelin and components of the reelin signaling pathway were measured by western blot and flow cytometry. Reelin inhibited the motility of control cells but not patient cells, and increased the number and size of focal adhesions in control cells but not patient cells. Patient and control cells expressed similar levels of the reelin receptors and the reelin signaling protein, Dab1, but patient cells expressed less reelin. Patient cells were smaller than control cells and had less actin and acetylated α-tubulin, components of the cytoskeleton. These findings are the first direct evidence that cellular responses to reelin are impaired in schizophrenia and are consistent with the role of reelin in cytoarchitectural deficits observed in schizophrenia patient brains.
在精神分裂症患者死后的大脑的各个区域,reelin 的表达减少,但 reelin 功能在精神分裂症的神经生物学中的确切作用仍然难以捉摸。由于神经元迁移异常,reelin 缺失的敲除小鼠导致新皮层的层状结构反转。本研究的目的是利用患者来源的嗅球源性(ONS)细胞来研究细胞外 reelin 是否会改变精神分裂症患者来源细胞的细胞迁移能力。将来自 9 名患者的 ONS 细胞与来自 9 名匹配的健康对照的细胞进行比较。使用自动化高通量成像和分析来跟踪在由 HEK293FT 细胞转染全长 reelin 质粒 pCrl 分泌到培养基中的 reelin 产生的 reelin 包被表面上的单个活细胞的迁移能力。使用自动测定法来量化细胞内细胞骨架组成、细胞形态和焦点粘连。通过 Western blot 和流式细胞术测量 reelin 和 reelin 信号通路的成分的表达。Reelin 抑制对照细胞的迁移,但不抑制患者细胞的迁移,并且增加对照细胞而非患者细胞中的焦点粘连的数量和大小。患者和对照细胞表达相似水平的 reelin 受体和 reelin 信号蛋白 Dab1,但患者细胞表达较少的 reelin。与对照细胞相比,患者细胞较小,并且细胞骨架的组成部分肌动蛋白和乙酰化α-微管蛋白较少。这些发现是细胞对 reelin 的反应受损在精神分裂症中的第一个直接证据,并且与在精神分裂症患者大脑中观察到的 reelin 在细胞结构缺陷中的作用一致。