A bacterial artificial chromosome transgenic mouse model for visualization of neurite growth

• Published in: Science China. Life sciences 2015-04, Vol.58 (4), p.373-378
• Main Authors: Tao, Tao, Chen, Chen, Sun, Jie, Peng, YaJing, Zhu, MinSheng
• Format: Article
• Language: English
• Subjects: Animals; Biomedical and Life Sciences; Chromosomes, Artificial, Bacterial; Life Sciences; Mice; Mice, Transgenic; Neurites; Recombinant Fusion Proteins - genetics; Research Paper; 可视化; 微管蛋白; 神经元发育; 红色荧光蛋白; 细菌人工染色体; 融合蛋白; 转基因小鼠模型; 轴突生长
• ISSN: 1674-7305; 1869-1889
• DOI: 10.1007/s11427-015-4820-9

Class Ⅲ β-tubulin （Tubb3） is a component of the microtubules in neurons and contributes to microtubule dynamics that are required for axon outgrowth and guidance during neuronal development. We here report a novel bacterial artificial chromosome （BAC） transgenic mouse line that expresses Class Ⅲ β-tubulin fused to mCherry, an improved monomeric red fluorescent protein, for the visualization of microtubules during neuronal development. A BAC containing Tubb3 gene was modified by insertion of mCherry complementary DNA downstream of Tubb3 coding sequence via homologous recombination, mCherry fusion protein was expressed in the nervous system and testis of the transgenic animal, and the fluorescent signal was observed in the neurons that located in the olfactory bulb, cerebral cortex, hippocampal formation, cerebellum, as well as the retina. Besides, Tubb3-mCherry fusion protein mainly distributed in neurites and colocalized with endogenous Class Ⅲ β-tubulin The fusion protein labels Purkinje cell dendrites during cerebellar circuit formation. Therefore, this transgenic line might be a novel tool for scientific community to study neuronal development both in vitro and in vivo.