Max Planck Institute of Biophysics, 60438 Frankfurt, Germany.
Cell. 2011 Nov 11;147(4):907-21. doi: 10.1016/j.cell.2011.09.049.
Each olfactory sensory neuron (OSN) in mouse chooses one of 1,200 odorant receptor (OR) genes for expression. OR genes are chosen for expression by greatly varying numbers of OSNs. The mechanisms that regulate the probability of OR gene choice remain unclear. Here, we have applied the NanoString platform of fluorescent barcodes and digital readout to measure RNA levels of 577 OR genes in a single reaction, with probes designed against coding sequences. In an inbred mouse strain with a targeted deletion in the P element, we find that this element regulates OR gene choice differentially across its cluster of 24 OR genes. Importantly, the fold changes of NanoString counts in ΔP or ΔH mice are in very close agreement with the fold changes of cell counts, determined by in situ hybridization. Thus, the P and H elements regulate the probability of OR gene choice, not OR transcript level per OSN.
在小鼠中,每个嗅觉感觉神经元(OSN)选择 1200 个气味受体(OR)基因中的一个进行表达。OR 基因的表达选择受到 OSN 数量的极大影响。调控 OR 基因选择概率的机制尚不清楚。在这里,我们应用荧光条码的 NanoString 平台和数字读取来测量单个反应中 577 个 OR 基因的 RNA 水平,使用针对编码序列设计的探针。在一个具有 P 元件靶向缺失的近交系小鼠中,我们发现该元件在其 24 个 OR 基因簇中差异调控 OR 基因的选择。重要的是,在ΔP 或ΔH 小鼠中 NanoString 计数的倍数变化与通过原位杂交确定的细胞计数的倍数变化非常吻合。因此,P 和 H 元件调控 OR 基因选择的概率,而不是每个 OSN 的 OR 转录本水平。