STOCK Tg(tetO-GCAMP2)12iRyu/J

Stock number: 017755
Research areas: Neurobiology Research, Research Tools

Description

These inducible calcium sensing GCaMP2 transgenic mice allow transcranial imaging of neurons in live animals.

Detailed description

This inducible calcium sensor transgene contains a conditional eGFP-calmodulin fusion protein, GCaMP2. Hemizygotes are viable and fertile, normal in size and do not display any gross physical or behavioral abnormalities. When bred with transgenic mice expressing a tetracycline transactivator (tTA) GCAMP2, expression is observed in cells of double mutant offspring. An increase of green fluorescence can be observed as the result of elevated intracellular calcium. For example, when bred to mice expressing the mouse olfactory marker protein (OMP) driving tTA (Stock No. 017754), GCAMP2 expression is localized in the vomeronasal neurons and main olfactory sensory neurons, which detect olfactory chemical stimuli. These chemical messages are detected by their binding to G protein-coupled receptors. The receptors activation elevates intracellular calcium, which results in an increase of fluorescence in specific neurons. These mice may be useful for slice preparation imaging or transcranial imaging of activated neurons in live animals.

Development

This transgene contains a calcium-sensing molecule, GCaMP2, under the transcriptional control of the tetracycline-responsive promoter element (tetO). GCaMP2 consists of, from 5' to 3', a 13-residue peptide of myosin light chain kinase (M13), a circularly permutated eGFP interrupted at residue 145, calmodulin (CaM), and a polyA recognition sequence. Mutations resulting in increased brightness (D180Y and V93I), thermal stability (V163A, and S175G) and GFP dimerization prevention (A206K) were introduced. This construct was injected into fertilized (CBAxFVB)F1 oocytes and mice from founder line 12i were bred to C57BL/6 mice to establish a colony. Transgenic offspring were bred to C57BL/6 for at least 3 generations. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Tg(tetO-GCaMP2)12IRyu
Name: transgene insertion 12I, Ron Yu
MGI accession ID: MGI:5301601
Generation method: Transgenic
Attributes: Reporter; Inducible
Marker symbol: Tg(tetO-GCaMP2)12IRyu
Marker name: transgene insertion 12I, Ron Yu
Chromosome: UN
Strain of origin: (CBA x FVB)F1
Expressed genes: GFP,Green Fluorescent Protein,; Mylk,myosin, light polypeptide kinase,mouse, laboratory; Calm1,calmodulin 1,mouse, laboratory; GCaMP,Genetically encoded calcium indicator,
Site of expression: When bred to mice expressing a tetracycline transactivator (tTA) or reverse transactivator (rtTA), doxycycline induction results in the expression of a GFP-calmodulin fusion protein, GCaMP2, in transactivator expressing tissues of the offspring.
Molecular note: This transgene contains a calcium-sensing molecule, GCaMP2, under the transcriptional control of the tetracycline-responsive promoter element (tetO). GCaMP2 consists of, from 5' to 3', a 13-residue peptide of myosin light chain kinase (M13), a circularly permutated eGFP interrupted at residue 145, calmodulin (CaM), and a polyA recognition sequence. Mutations resulting in increased brightness (D180Y and V93I), thermal stability (V163A, and S175G), and GFP dimerization prevention (A206K) were introduced.