概括
鼠类红血红血病 (MEL) 细胞是用人类β-环球蛋白基因进行的. 化学诱导剂激活了这些基因,在红色球体分化过程中显示了人类和小鼠细胞之间保留的调节序列.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 细胞生物学 细胞生物学
背景情况:
- 鼠类红血球瘤 (MEL) 细胞是研究红细胞分化的一个有价值的模型.
- 了解全球蛋白基因表达的调节对于血液学研究至关重要.
研究的目的:
- 调查MEL细胞中外源人类和混合β-环球蛋白基因的调节.
- 为了确定全球因基因表达的调节序列是否在人类和小鼠之间得到保护.
主要方法:
- 在MEL细胞中引入混合小鼠-人类和完整的人类β-环球蛋白基因.
- 使用化学剂诱导差异化.
- 使用S1核酶和原始延伸试验分析mRNA水平.
- 通过核转录实验评估全球蛋白基因转录率.
主要成果:
- 化学诱导剂增加了外源性全球蛋白基因mRNA水平的5到50倍.
- 诱导的mRNAs表现出正确的启动和终止.
- 全球蛋白基因诱导是特定于红色素细胞的.
- 转录率的增加有助于混合mRNA的诱导.
结论:
- 特定诱导β-环球蛋白基因是它们的DNA序列的固有属性.
- 对于β-环球蛋白基因表达的调节序列在人类和小鼠之间保持着保留.
- MEL细胞提供了一个系统,用于研究红状腺分化过程中外源性全球蛋白基因调节.
相关概念视频
09:21Phenotypic Profiling of Human Stem Cell-Derived Midbrain Dopaminergic Neurons
2.0K
This protocol describes the cell culturing of human midbrain dopaminergic neurons, followed by immunological staining and the generation of neuronal phenotypic profiles from acquired microscopic high-content images allowing the identification of phenotypic variations due to genetic or chemical...
2.0K
10:54Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression
10.0K
In Parkinson's Disease (PD), Substantia Nigra (SNc) dopaminergic neurons degenerate, leading to motor dysfunction. Here we report a protocol for culturing ventral midbrain neurons from a mouse expressing eGFP driven by a Tyrosine Hydroxylase (TH) promoter sequence, harvesting individual fluorescent neurons from the cultures, and measuring their transcriptome using...
10.0K
08:45Isolation, Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains
14.5K
The causes of degeneration of midbrain dopaminergic neurons during Parkinson’s disease are not fully understood. Cellular culture systems provide an essential tool for study of the neurophysiological properties of these neurons. Here we present an optimized protocol, which can be utilized for in vitro modeling of...
14.5K
09:35Environmental Modulations of the Number of Midbrain Dopamine Neurons in Adult Mice
9.2K
This protocol describes two different environmental manipulations and a concurrent brain infusion protocol to study environmentally-induced brain changes underlying adaptive behavior and brain repair in adult...
9.2K
11:58Primary Culture of Mouse Dopaminergic Neurons
39.1K
Dopaminergic neurons play a vital regulatory role in the brain. Their loss is associated with Parkinson's disease. In this video, we show how to generate primary cultures of central dopaminergic neurons from embryonic mouse mesencephalon. Such cultures are useful to study the extreme vulnerability of these neurons to various...
39.1K
09:54Comprehensive Profiling of Dopamine Regulation in Substantia Nigra and Ventral Tegmental Area
26.4K
Dopamine is distinctly regulated in the midbrain nuclei, which contain the cell bodies and dendrites of the dopamine neurons. Here we describe a dissection and sample-handling approach to maximize results, and thus conclusions and insights, on dopamine regulation in the midbrain nuclei of the substantia nigra (SN) and ventral tegmental area (VTA) in...
26.4K


