Transgenic mice expressing a dual, CRE-inducible reporter for the analysis of axon guidance and synaptogenesis.
Improved and modular tools are needed for the neuroanatomical dissection of CNS axonal tracts, and to study the cell-intrinsic and cell-extrinsic cues that govern their assembly and plasticity. Here we describe a general purpose transgenic tracer that can be used to visualize axonal tracts and synaptic terminals in any region of the embryonic neural tube or postnatal CNS, on any wild type or mutant genetic background. The construct permits CRE-inducible expression of a dicistronic axonal marker encoding two surface reporter proteins: a farnesylated GFP and the human Placental Alkaline Phosphatase (PLAP). Both proteins localize alongside the neuronal surface, permitting the concomitant detection of cell body, neurites, and presynaptic and postsynaptic sites in the same neuron. This provides a CRE-inducible dual system for imaging neural circuits in vivo, and to study their assembly and remodeling in cultured neurons, neural stem cells, and tissue explants derived from the reporter line. Unlike existing lines, this reporter does not encode a ubiquitously expressed, floxable LacZ gene, permitting the simultaneous analysis of beta galactosidase activity in mutant lines.
Badaloni
Aurora
A
San Raffaele Scientific Institute and Università Vita-Salute San Raffaele, Milano, Italy.
Bonanomi
Dario
D
Albieri
Ilaria
I
Givogri
Irene
I
Bongarzone
Ernesto
E
Valtorta
Flavia
F
Consalez
G Giacomo
GG
D000469:Alkaline Phosphatase / Q000032:analysis* / Q000235:genetics; D000818:Animals; D001369:Axons / Q000737:chemistry / Q000502:physiology*; D002490:Central Nervous System / Q000737:chemistry / Q000196:embryology / Q000254:growth & development*; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D019426:Integrases / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic / Q000196:embryology / Q000235:genetics / Q000254:growth & development*; D009474:Neurons / Q000737:chemistry / Q000502:physiology; D013569:Synapses / Q000737:chemistry / Q000502:physiology*; D019076:Transgenes
D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases; D000469:Alkaline Phosphatase