Rothman Andrea, Feinstein Paul, Hirota Junji, Mombaerts Peter
The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
Mol Cell Neurosci. 2005 Mar;28(3):535-46. doi: 10.1016/j.mcn.2004.11.006.
From a repertoire of approximately 2000 odorant receptor (OR) alleles in the mouse genome, a mature olfactory sensory neuron (OSN) is thought to choose only one functional allele of one OR gene for expression. OSNs that express a given OR gene are scattered throughout an epithelial region that is gene specific. The DNA sequences that enable OR gene choice and specify the epithelial pattern are not known. Within the upstream regions of several mouse, rat, and human OR genes, we have previously recognized putative homeodomain and O/E-like binding sites in proximity to each other. Here, we define a minimal promoter region for expression of the mouse OR gene M71 with small transgenes. This region contains a homeodomain and an O/E-like binding site. Combined mutations in both sites abolish transgene expression. When identical mutations are introduced at the endogenous M71 locus by gene targeting, the number of M71-expressing OSNs is reduced by a factor of three and the epithelial pattern is ventralized. The stronger impact observed with the mutant transgenes compared to the targeted mutations may reflect a multiplicity of regulatory sites within the OR gene cluster. We propose that these homeodomain and O/E sites regulate the probability of M71 gene choice differentially across the olfactory epithelium.
在小鼠基因组中约2000个气味受体(OR)等位基因的库中,一个成熟的嗅觉感觉神经元(OSN)被认为仅选择一个OR基因的一个功能等位基因进行表达。表达给定OR基因的OSN分散在一个基因特异性的上皮区域中。能够实现OR基因选择并确定上皮模式的DNA序列尚不清楚。在几个小鼠、大鼠和人类OR基因的上游区域内,我们之前已经识别出彼此相邻的假定同源结构域和O/E样结合位点。在这里,我们用小转基因定义了小鼠OR基因M71表达的最小启动子区域。该区域包含一个同源结构域和一个O/E样结合位点。两个位点的联合突变消除了转基因表达。当通过基因靶向在内源M71位点引入相同的突变时,表达M71的OSN数量减少了三分之一,并且上皮模式向腹侧化。与靶向突变相比,在突变转基因中观察到的更强影响可能反映了OR基因簇内多个调控位点。我们提出,这些同源结构域和O/E位点在整个嗅觉上皮中差异调节M71基因选择的概率。