实验方法> 生物化学技术> 化学生物学实验技术>A Genetic Strategy for the Analysis of Individual Axon Morphologies in cGMP Signalling Mutant Mice

A Genetic Strategy for the Analysis of Individual Axon Morphologies in cGMP Signalling Mutant Mice

关键词: genetic strategy analysis来源: 互联网

One of the many physiological functions of cyclic guanosine 3′,5′ monophosphate (cGMP) signalling is the regulation of a specific mode of axonal branching. The bifurcation of axons from dorsal root ganglion (DRG) neurons at the dorsal root entry zone of the embryonic spinal cord is triggered by a cGMP �signalling pathway comprising the ligand C-type natriuretic peptide (CNP), the cGMP-producing natriuretic peptide receptor 2 (Npr2), and the cGMP-dependent protein kinase Iα (cGKIα). Absence of any of these components causes a loss of bifurcation and sensory axons instead only turn in either a rostral or a caudal direction. In this chapter we describe a genetic strategy to study the impact of cGMP signalling on the arborization of individual DRG neurons in mice. Expression of an alkaline phosphatase (AP) reporter is selectively induced in Npr2-positive DRG neurons by tamoxifen-dependent activation of a Cre �recombinase under the control of the Npr2 promoter. This approach might also be employed for the analysis of axonal branching in neuronal subsets expressing Npr2 elsewhere in the nervous system.

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