中国科学院昆明动物研究所机构知识库

人脑中的一种特别的双极神经元VEN在认知，自我意识，情感，方面可能有非常重要的作用。VEN数量和体积在灵长类物种与这些物种的认知程度呈高度正相关，表明VEN在灵长类认知，自我意识，情感的进化起到关键作用，是高级认知，智力，情感进化的产物。到目前为止，我们对VEN形态和功能的内在生物学机制依然知之甚少。为了加速VEN的功能研究，我们通过转录组测序提供了一个VEN的转录组表达数据，为后续的VEN功能机制研究创造条件。为了能够充分地探究VEN形态和功能的内在分子机制，探索VEN的演化，我们通过激光显微切割技术收集了人前扣带皮层BA24区第五层VEN，第三层锥体神经元，第五层锥体神经元和前扣带皮层BA32区第五层锥体神经元，以及猕猴和食蟹猴前脑岛第五层VEN，第五层锥体神经元，第三层锥体神经元进行转录组测序和分析。通过比较人四种神经元类型，我们识别了215个人VEN特异高表达基因，129个人VEN特异低表达基因。这些基因的功能主要与胞体，树突形态，髓鞘形成，中间纤维组织等有关，通过蛋白质互作网络分析，我们识别了参与食物摄取，焦虑，压力，痛觉与节律，神经突发生，髓鞘和神经纤维形成的基因，如NPY，HPCA，HPCAL1，RALGDS，NEFH，MBP和PLP1等。我们通过原位杂交、免疫组化和免疫荧光证实VAT1L， LYPD1，SULF2和CHST8在VEN中特异高表达，可以作为VEN markers。NEFH和VGLUT1在VEN中高表达，说明VEN是兴奋性投射神经元。我们也分别识别了食蟹猴和猕猴VEN特异高表达基因。食蟹猴VEN特异高表达基因在细胞命运决定，智力，神经突生长和抑制，行为等发面有重要作用。猕猴特异高表达基因参与了一些神经退行性疾病和认知疾病的发生，如阿尔兹海默症，脊髓小脑共济失调，智力缺陷，自闭症，癫痫等。通过分析人，猕猴和食蟹猴同源基因系统发生树发现，VEN按照物种聚类，表明种间差异要大于神经元类型差异。通过网络分析发现，60%以上的三个物种共享的VEN高表达同源基因有蛋白质相互作用，整个网络中80%以上的基因来自三个物种共享的VEN高表达同源基因，这些基因在转录及转录后修饰，翻译及翻译后修饰，囊泡运输，细胞骨架等层面调控VEN形态发生和功能形成，反映了人，猕猴和食蟹猴VEN最基本的分子属性和共有特征。总结来说，我们通过转录组数据全面系统地展现了人，猕猴和食蟹猴VEN的表达图景，识别了一些决定VEN形态和功能的基因，找到了人VEN特异markers，阐释了人，食蟹猴和猕猴VEN的系统发生关系，鉴定了与VEN基本属性有关的基因。

A very important and special bipolar spindle neuron called von Economo neuron (VEN) may play very important roles in cognition, self-awareness, emotion, The number and volume of VENs in primate species are highly positively correlated with the cognitive level of these species, suggesting VENs play key roles in the evolution of cognition, self-awareness and emotion in primates and the products of evolution of advanced cognition, intelligence, emotion.We still know little about the underlying biological mechanism of VEN morphogenesis and function although previous research has found some VEN-related genes up to now. In order to speed up the functional research of VENs, we provided a transcriptome expression data of VENs through transcriptome sequencing to set the stage for subsequent functional studies of VENs. In order to fully explore the evolution of VEN and the molecular mechanism of VEN morphogenesis and function, we collected VENs，layer Vb pyramidal neurons, layer III pyramidal neurons from BA24 of the human anterior cingulate cortex , cynomolgus monkey and rhesus monkey anterior insula cortex respectively, we also collected the layer Vb pyramidal neurons from human BA32 of the anterior cingulate cortex as a reference cell type. The collected neuron samples were subject to RNA sequencing. By comparing among human four cell types, we identified 215 VEN-specific highly expression genes and 129 VEN-specific lowly expression genes that were involed in cell body, dendritic morphology, myelin formation and intermediate fiber organization. We identified the genes involved in food intake, anxiety, stress, pain and rhythm, neurite outgrowth, myelin formation and nerve fiber formation by protein-protein interaction, such as NPY, HPCA, HPCAL1, RALGDS, NEFH, MBP and PLP1. We validated VAT1L, LYPD1, SULF2 and CHST8 were the VEN-specific highly expression genes by mRNA in situ hybridization, immunocytochemistry and Immunofluorescence, and they could be served as the VEN markers. High expression for VGLUT1 and NEFH in human VENs suggested that VENs were excitatory projection neurons. Moreover, giving the importance of high expression genes, we also identified respectively cynomolgus macaque and rhesus macaque VEN-specific highly expression genes, which play important roles in cell fate determination, intelligence, neurite outgrowth, depression, behavior, onset and progress of neurodegenerative and cognitive diseases such as Alzheimer's disease, spinocerebellar ataxia, mental retardation, autism and epilepsy. According to the homologous genes phylogenetic tree among human, cynomolgus macaque and rhesus macaque, it was found that VENs were clustered according to species, indicating that the differences between species were greater than the differences of neuron types for VENs, layer Vb pyramidal neurons and layer III pyramidal neurons. Moreover, over 60% of the three species-shared VENs highly expressed homologous genes(TSVHG) involed in protein-protein interaction network, and over 80% of the genes in the network were from TSVHG which regulated VEN morphogenesis and fuction on the levels of transcription and post-transcriptional modification, translation and post-translational modification, vesicular transport and cytoskeleton organization. It reflected the basic molecular properties and common features of VENs from human, cynomolgus macaque and rhesus macaque. Taken together, we systematically presented the expression profile of VENs from human, cynomolgus macaque and rhesus macaque, identified some genes that determined the morphology and function of VENs, found the human VEN-specific markers, stated phylogenetic relationship among human, cynomolgus macaque and rhesus macaque VENs, and identified the genes related to the basic properties of VENs.