Microarray and real-time PCR analyses of gene expression in the honeybee brain following caffeine treatment
Microarray and real-time PCR analyses of gene expression in the honeybee brain following caffeine treatment
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DOI：
10.1385/jmn:27:3:269
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发表时间：
2005-01-01
影响因子：
3.1
通讯作者：
Maleszka, R
中科院分区：
文献类型：
--
作者：
Kucharski, R;Maleszka, R
为了验证咖啡因可能诱导蜜蜂大脑中基因表达变化的想法，我们使用高通量cDNA微阵列对比了对照组和咖啡因处理组大脑的转录谱。对八个转录本的子集进行额外的定量实时PCR，以可视化咖啡因诱导的时间变化。在咖啡因处理的大脑中显著上调的基因包括那些参与突触信号传导（GABA：Na同向转运体、多巴酚丁胺D2R样受体和突触蛋白）、细胞骨架修饰（驱动蛋白和微管马达）、蛋白质翻译（核糖体蛋白RpL4、延伸因子）和钙依赖性过程（钙转运蛋白、钙调蛋白依赖性环核苷酸磷酸二酯酶）的基因。此外，我们的研究还发现了一些新的咖啡因诱导基因，这些基因似乎是蜜蜂所独有的。咖啡因敏感基因表达的时间依赖性分析显示，治疗后1小时显著上调，4小时后中度下调，24小时后无其他变化。我们的研究结果提供了初步的证据表明，多巴胺能系统和钙离子交换的主要目标咖啡因在蜜蜂的大脑，并建议在无脊椎动物的大脑中的分子反应咖啡因是类似的脊椎动物生物体。
To test the idea that caffeine might induce changes in gene expression in the honeybee brain, we contrasted the transcriptional profiles of control and caffeine-treated brains using high-throughput cDNA microarrays. Additional quantitative real-time PCR was performed on a subset of eight transcripts to visualize the temporal changes induced by caffeine. Genes that were significantly upregulated in caffeine-treated brains included those involved in synaptic signaling (GABA:Na symporter, doparnine D2R-like receptor, and synapsin), cytoskeletal modifications (kinesin and microtubule motors), protein translation (ribosomal protein RpL4, elongation factors), and calcium-dependent processes (calcium transporter, calmodulin-dependent cyclic nucleotide phosphodiesterase). In addition, our study uncovered a number of novel, caffeine-inducible genes that appear to be unique to the honeybee. Time-dependent profiling of caffeine-sensitive gene expression shows significant upregulation 1 h after treatment followed by moderate downregulation after 4 h with no additional changes occuring after 24 h. Our results provide initial evidence that the doparninergic system and calcium exchange are the main targets of caffeine in the honeybee brain and suggest that molecular responses to caffeine in an invertebrate brain are similar to those in vertebrate organisms.