Takamatsu Yoshiki, Nakagoshi Hideki, Rachidi Mohammed, Lopes Carmela, Nishida Yasuyoshi, Ohsako Shunji
Department of Brain Structure, Tokyo Metropolitan Institute for Neuroscience, 2-6 Musashidai, Fuchu, Tokyo 183-8526, Japan.
Cell Tissue Res. 2002 Nov;310(2):237-52. doi: 10.1007/s00441-002-0631-y. Epub 2002 Sep 28.
Transgenic flies that can drive GAL4 expression under the control of the 7 kb 5'-region of the Drosophila Ca(2+)/calmodulin-dependent protein kinase II (dCaMKII) gene (dCaMKII-GAL4) were established. Characteristic features of this dCaMKII-GAL4 driven reporter expression were compatible with the endogenous dCaMKII expression pattern: The dCaMKII-GAL4 driven reporter gene was expressed preferentially in the central nervous system of the embryo and larvae. Reporter expression was also observed in the brain, thoracic ganglion, and gut of the adult. The whole-brain distribution and projections of dCaMKII-GAL4-expressing cells in the adults were visualized three-dimensionally by using UAS-linked reporter genes. Prominent signals of nuclear-localized beta-Gal reporter gene expression were found in extensive brain regions, especially in the Kenyon cells of the mushroom body (MB), cells in the pars intercerebralis, and subesophageal ganglion (SOG). tau reporter gene expression highlighting neurite projections was detected in the MB lobes, median bundle, antennal lobe glomeruli, and fibers of clusters in the SOG, ventrolateral protocerebrum and superior lateral protocerebrum. These observations agree with those of a previous study mapping the dCaMKII-dependent memory circuits in courtship conditioning. Interestingly, green fluorescent protein reporter gene expression in adult MB lobes was predominantly observed in the alpha and beta lobes with a core-deficient pattern, but not in the alpha' and beta' lobes, similar to Fasciclin II immunoreactivity.
构建了能够在果蝇钙/钙调蛋白依赖性蛋白激酶II(dCaMKII)基因7 kb 5'-区域控制下驱动GAL4表达的转基因果蝇(dCaMKII-GAL4)。这种由dCaMKII-GAL4驱动的报告基因表达的特征与内源性dCaMKII表达模式相符:dCaMKII-GAL4驱动的报告基因在胚胎和幼虫的中枢神经系统中优先表达。在成虫的脑、胸神经节和肠道中也观察到报告基因的表达。通过使用与UAS相关的报告基因,对成虫中表达dCaMKII-GAL4的细胞的全脑分布和投射进行了三维可视化。在广泛的脑区,特别是在蘑菇体(MB)的肯扬细胞、脑间部的细胞和咽下神经节(SOG)中发现了核定位β-半乳糖苷酶报告基因表达的显著信号。在MB叶、中束、触角叶小球以及SOG、腹侧原脑和上外侧原脑的簇状纤维中检测到突出神经突投射的tau报告基因表达。这些观察结果与先前一项在求偶条件反射中绘制dCaMKII依赖性记忆回路的研究结果一致。有趣的是,在成虫MB叶中绿色荧光蛋白报告基因的表达主要在α和β叶中以核心缺失模式观察到,而在α'和β'叶中未观察到,这与Fasciclin II免疫反应性相似。