| Optogenetic Mapping of Synaptic Connections in Mouse Brain Slices to Define the Functional Connectome of Identified Neuronal Populations
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Author:
Date:
2017-01-05
[Abstract] Functional connectivity in a neural circuit is determined by the strength, incidence, and neurotransmitter nature of its connections (Chuhma, 2015). Using optogenetics the functional synaptic connections between an identified population of neurons and defined postsynaptic target neurons may be measured systematically in order to determine the functional connectome of that identified population. Here we describe the experimental protocol used to investigate the excitatory functional connectome of ventral midbrain dopamine neurons, mediated by glutamate cotransmission (Mingote et al., 2015). Dopamine neurons are made light sensitive by injecting an adeno-associated virus (AAV) encoding channelrhodopsin (ChR2) into the ventral midbrain of DATIREScre mice. The efficacy and ...
[摘要] 神经回路中的功能连通性由其连接的强度，发病率和神经递质特性决定（Chuhma，2015）。使用光遗传学，可以系统地测量识别的神经元群体和定义的突触后靶神经元之间的功能性突触连接，以便确定该识别群体的功能性连接群。这里我们描述了用于研究由谷氨酸共转播介导的腹侧中脑多巴胺神经元的兴奋性功能性连接体的实验方案（Mingote等，2015）。通过将编码通道视紫红质（ChR2）的腺相关病毒（AAV）注射到DATIREScre小鼠的腹侧中脑中，使多巴胺神经元变得光敏。多巴胺合成酶酪氨酸羟化酶的免疫荧光证实ChR2表达在多巴胺神经元中的功效和特异性。然后，切片膜片钳记录由接受多巴胺神经元投影的区域中的神经元产生，并且确定兴奋性连接的发生率和强度。所有接受多巴胺神经元投射的区域的连接发生率和强度的总结构成功能性连接体。
【背景】为了建立特定神经回路的功能，有必要确定解剖连接，解剖连接的映射及其功能连接，连接的强度，发病率和神经递质性质的映射。使用单因素限制的病毒性突触后追踪技术，可以描述包括多巴胺系统在内的神经回路的复杂解剖连接（Callaway and Luo，2015; ...
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| Estimation of the Chromosomal Copy Number in Synechococcus elongatus PCC 7942
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Author:
Date:
2016-07-05
[Abstract] Cyanobacteria are prokaryotic organisms that perform oxygenic photosynthesis. Freshwater cyanobacteria, such as Synechococcus elongatus PCC 7942 and Synechocystis sp. PCC 6803, are model organisms for the study of photosynthesis, gene regulation, and biotechnological applications because they are easy to manipulate genetically. However, while studying these cyanobacteria, care has to be taken with respect to genetic heterogeneity in the establishment of gene disruptants, because these cyanobacteria contain multiple chromosomal copies per cell. Here, we describe a method for the estimation of chromosomal copy number in Synechococcus 7942. Using this method, we have recently observed that the chromosomal copy number of Synechococcus 7942 ...
[摘要] 蓝细菌是进行含氧光合作用的原核生物。 淡水蓝细菌，例如细长聚球藻PCC 7942和聚集细胞 PCC 6803是用于研究光合作用，基因调节和生物技术应用的模式生物，因为它们易于基因操作。 然而，当研究这些蓝细菌时，必须注意遗传异质性在基因破坏物的建立中，因为这些蓝细菌每个细胞含有多个染色体拷贝。 在这里，我们描述了用于估计在聚球藻7942中的染色体拷贝数的方法。使用这种方法，我们最近观察到，聚球藻7942的染色体拷贝数显着地改变 其生长阶段。 这种技术可用于研究多倍体不仅在蓝细菌，而且在其他多倍体生物。
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| Estradiol Receptor (ER) Chromatin Immunoprecipitation in MCF-7 Cells
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Author:
Date:
2013-07-20
[Abstract] Steroid hormone receptors, for example estradiol receptor, act like transcription factors. In the cell, steroids bind to a specific receptor. Upon ligand binding, many steroid receptors dimerize and enter nuclei where they bind specific DNA sequences called Hormone Responsive Elements (HRE) and regulate gene transcription. ER is able to bind DNA sites that are not Estrogen Responsive Elements (ERE) so regulating also the transcription of genes that are not classically controlled by estrogens.
[摘要] 类固醇激素受体，例如雌二醇受体，像转录因子一样起作用。 在细胞中，类固醇与特异性受体结合。 在配体结合时，许多类固醇受体二聚化并进入细胞核，其中它们结合称为激素应答元件（HRE）的特异性DNA序列并调节基因转录。 ER能够结合不是雌激素反应元件（ERE）的DNA位点，从而也调节不经典地由雌激素控制的基因的转录。
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