Mapping of the Developmental Atlas of the Visual System and Research on Embryonic Neurogenesis Phenomena
试验速览
- 阶段
- 不适用
- 状态
- 招募中
- 发起方
- 入组人数
- 200
- 试验地点
- 1
- 主要终点
- Single-cell transcriptomic atlas and developmental trajectory of embryonic retinal and visual cortical neurons
研究概览
简要总结
This research studies how nerve cells in the human embryonic retina, visual brain regions, and brain areas responsible for higher cognitive functions grow, develop, and form interconnected functional networks.
Eye tissue, visual brain tissue, and other brain tissue linked to advanced cognitive functions will be collected from embryos whose pregnancies were terminated due to medical conditions or illnesses. High-throughput single-cell and single-nucleus sequencing will be utilized to map gene activity patterns and developmental growth pathways of retinal nerve cells.
Multiple testing tools will be combined to analyze these brain and retinal cells: Patch-seq (single-cell patch-clamp sequencing), high-density microelectrode arrays (MEA), and two-photon calcium imaging. With these tools, systematic measurements will be performed on the electrical activity, physical shape, synaptic connection patterns, and signal coding functions of neurons in the retina and visual brain regions. Multi-modal tissue maps and a public database will be constructed to store all collected research data.
Immunofluorescence staining will also be applied to compare structural differences and nerve fiber connections between visual brain regions and higher cognitive brain areas. The regenerative capacity and neuron formation process of embryonic brain stem cells, as well as the migration paths of developing neurons, will be tracked.
Overall, this study aims to fully uncover the neural foundation of visual signal processing, and identify the molecular regulatory networks that control nerve tissue development during the embryonic stage.
研究设计
- 研究类型
- Observational
- 观察模型
- Case Control
- 时间视角
- Retrospective
入排标准
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Abnormal group: Embryos with clinically confirmed embryonic developmental abnormalities requiring medical termination of pregnancy; intact retinal, visual and cognitive brain tissues available for snATAC-seq, scRNA-seq, electrophysiology, proteomics and RNAscope detection.
- •Normal group: Embryos confirmed free of any ocular and central nervous developmental defects by prenatal examination and anatomical observation; intact embryonic ocular and brain tissues meeting all experimental detection standards.
- •All sample donors have signed written informed consent authorizing the use of residual embryonic tissues for scientific research, with no monetary compensation involved.
排除标准
- •Embryonic ocular or brain tissues with severe necrosis, structural damage or microbial contamination that cannot support multi-omics and functional experiments.
- •Donors who withdraw or refuse the consent for tissue research use. Samples with irregular collection, transportation or cryopreservation procedures resulting in tissue degradation and failure to meet experimental requirements.
结局指标
主要结局
Single-cell transcriptomic atlas and developmental trajectory of embryonic retinal and visual cortical neurons
时间窗: Day 1 of tissue collection
Perform single-cell RNA sequencing on human embryonic eye and brain tissues from abnormal developmental group and normal control group. Identify all cell subtypes in retina and visual-associated cognitive cortex, reconstruct continuous developmental trajectory of retinal neurogenesis and visual cortical neurons, and compare neuronal subtype distribution differences between the two groups.
Genome-wide chromatin open regions in embryonic visual tissues
时间窗: Day 1 of tissue collection
Genome-wide chromatin open regions are detected via snATAC-seq on embryonic ocular and brain tissues from case and control groups.
Differential chromatin accessibility between normal and malformed embryonic visual tissues
时间窗: Day 1 of tissue collection
Differential chromatin accessibility signals are identified via integrated analysis of snATAC-seq and scRNA-seq data from embryonic ocular and brain tissues.
Candidate pathogenic genes underlying embryonic visual developmental abnormalities
时间窗: Day 1 of tissue collection
Genes linked to abnormal embryonic visual development are screened based on differential chromatin and transcriptomic profiles.
次要结局
- Action potential firing patterns of embryonic visual neurons(Day 1 of tissue collection)
- Synchronous electrical activity of embryonic neuronal networks(Day 1 of tissue collection)
- Spatial expression localization of key visual development genes detected by RNAscope(The day 1 of embryonic tissue collection after clinical termination of pregnancy)
- Differentially expressed proteins identified via global proteome sequencing(Day 1 of tissue collection)
- Spatial localization and expression abundance of proteins via immunohistochemistry(Day 1 of tissue collection)
研究者
Sheng Liu
Professor
Zhongshan Ophthalmic Center, Sun Yat-sen University