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

Abstract

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.

Nerve Growth Factor Secretion From Pulp Fibroblasts is Modulated by Complement C5a Receptor and Implied in Neurite Outgrowth

“…1Ae–g ). Nerve fibers were localized in the same tooth sections using ß-III-tubulin antibody, a marker of central and peripheral nerve fibers 22 . Uniform and scattered ß-III-tubulin expression was detected throughout the intact section ( Fig.…”

Pulp Fibroblasts Control Nerve Regeneration through Complement Activation

“…2G, H). Nerve fiber localization was investigated on the same tooth sections through β-III-tubulin staining, a differentiated neuron marker (Tao et al 2015). In intact teeth, β-III-tubulin was mainly localized beneath the odontoblast cell layer (Fig.…”