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High-Throughput Microarray Detection of Vomeronasal Receptor Gene Expression in Rodents

机译:高通量微阵列检测啮齿类动物的犁鼻受体基因表达

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We performed comprehensive data mining to explore the vomeronasal receptor (V1R & V2R) repertoires in mouse and rat using the mm5 and rn3 genome, respectively. This bioinformatic analysis was followed by investigation of gene expression using a custom designed high-density oligonucleotide array containing all of these receptors and other selected genes of interest. This array enabled us to detect the specific expression of V1R and V2Rs which were previously identified solely based on computational prediction from gene sequence data, thereby establishing that these genes are indeed part of the vomeronasal system, especially the V2Rs. 168 V1Rs and 98 V2Rs were detected to be highly enriched in mouse vomeronasal organ (VNO), and 108 V1Rs and 87 V2Rs in rat VNO. We monitored the expression profile of mouse VR genes in other non-VNO tissues with the result that some VR genes were re-designated as VR-like genes based on their non-olfactory expression pattern. Temporal expression profiles for mouse VR genes were characterized and their patterns were classified, revealing the developmental dynamics of these so-called pheromone receptors. We found numerous patterns of temporal expression which indicate possible behavior-related functions. The uneven composition of VR genes in certain patterns suggests a functional differentiation between the two types of VR genes. We found the coherence between VR genes and transcription factors in terms of their temporal expression patterns. In situ hybridization experiments were performed to evaluate the cell number change over time for selected receptor genes.
机译:我们进行了全面的数据挖掘,以分别使用mm5和rn3基因组来研究小鼠和大鼠中的犁鼻受体(V1R和V2R)组成。该生物信息学分析之后,使用定制设计的高密度寡核苷酸阵列研究基因表达,该阵列包含所有这些受体和其他感兴趣的选定基因。该阵列使我们能够检测以前仅根据基因序列数据的计算预测确定的V1R和V2R的特定​​表达,从而确定这些基因确实是犁鼻系统的一部分,尤其是V2R。检测到168个V1R和98个V2R在小鼠犁鼻器器官(VNO)中高度富集,在大鼠VNO中有108个V1R和87 V2Rs。我们监测了小鼠VR基因在其他非VNO组织中的表达谱,结果基于其非嗅觉表达模式,将某些VR基因重新指定为VR样基因。表征小鼠VR基因的时间表达谱并对其模式进行分类,揭示了这些所谓的信息素受体的发育动态。我们发现了许多时间表达模式,这些模式表明可能的行为相关功能。 VR基因在某些模式下的组成不均提示这两种VR基因之间存在功能差异。我们发现VR基因和转录因子之间的时间表达方式的一致性。进行原位杂交实验以评估所选受体基因随时间变化的细胞数。

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