B10

HCN4-GCaMP8 Strain Information Sheet

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(PDF mouse profile sheet HCN4-GCaMP8)

Strain Name:

Common Name: HCN4-GCaMP8

CHROMusTM designation and lines available: R24:B10:L3

Jackson Stock number: NA

Development: The green fluorescent calcium indicator GCaMP8 was inserted at the ATG site of the HCN4 gene in the BAC RP23-281H22 (CHORI) through homologous recombination. The resulting recombinant BAC was injected into the male pronucleus of fertilized oocytes which were then implanted into pseudo pregnant females. Resulting offspring were screened for the presence of the transgene (founders). Colonies were established from each founder and tested for expression.

Transgenic Numbers:

# pups born # founders # expressors
11 5 1

Description: The genetically encoded calcium indicator GCaMP8 is expressed under control of the HCN4 promoter, directing expression to the SA node and the AV node of the heart and the nervous system (brain and spinal cord). Expression is observed in the developing 13.5 dpc heart and in the brain and spinal cord. GCaMP8 responds to calcium levels in the cell. When calcium increases, a conformation change occurs resulting in an increase in fluorescence. When calcium decreases, fluorescence decreases. This mouse is useful for examining calcium signaling in the SA and AV nodes.

Phenotypic Data:

Figure 1: Native fluorescent images (GCaMP8) in 13 dpc embryo (left panel, genotypically negative, right panel genotypically positive) showing expression in the forebrain and spinal cord.

Figure 1: Native fluorescent images (GCaMP8) in 13.5 dpc embryo (left panel, genotypically negative, right panel genotypically positive) showing expression in the forebrain and spinal cord.

 

 

 

 

 

 

 

 

Figure 2: Native fluorescence (GCaMP8) in the heart of a 13 dpc embryo. Note the atrial signaling in the positive embryo.

Figure 2: Native fluorescence (GCaMP8) in the heart of a 13.5 dpc embryo. Note the atrial signaling in the positive embryo.

 

 

 

 

 

 

 

 

 

 

 

Figure 3: anti-gfp staining in the heart of a 13 dpc embryo. Whole embryo and heart showing staining in the SA node.

Figure 3: anti-gfp staining in the heart of a 13.5 dpc embryo. Whole embryo and heart showing staining in the SA node.

 

 

 

 

 

 

 

Figure 4:  calcium signaling in the SA node of a 13.5 dpc embryo.

 

Genotyping Protocol:

Primer CalmF AAG GGC GAG GAG CTG TTC A
Primer CalmR CGA TCT GCT CTT CAG TCA GTT GGT
Expected size of product: 456bp
Cycling conditions:

  1. 94oC 3 minutes
  2. Repeat 30 times
    1. 30 sec @ 94oC
    2. 30 sec @ 57oC
    3. 60 sec @ 72oC
  3. 72oC 5 minutes
  4. 10oC hold

PCR results:

GCaMP PCR for strain info

 

Terms of Use: Please contact chromus@cornell.edu for information on MTA and fees.

So as to appropriately acknowledge and credit the scientists who have produced the HCN4-GCaMP8 mouse strain, it is expected that you will include these individuals in any publication that is submitted prior to the initial publication of this line from the Kotlikoff laboratory.

Acknowledgement: Please inform us of any publications resulting from the use of this mouse and use the following statement to acknowledge their development:

The mouse strain HCN4-GCaMP8 was developed by CHROMusTM which is supported by the National Heart Lung Blood Institute of the National Institute of Health under award number R24HL120847.

 

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