B4

αMHC-GCaMP8 Strain Information Sheet

request mice

Strain Name:

Common Name: αMHC-GCaMP8

CHROMusTM designation and lines available: R24:B4

Jackson Stock number: NA

Development: The green fluorescent calcium indicator GCaMP8 was inserted under control of the αMHC minimal promoter (Addgene 55594). The resulting transgene 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 # expressers
20 3 3

Description: The genetically encoded calcium indicator GCaMP8 is expressed under control of the αMHC promoter, directing expression to cardiomyocytes in the heart. 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 cardiomyocytes.

Phenotypic Data:

qPCR analysis of GCAMP8 message in the heart.

qPCR analysis of GCAMP8 message in the heart.

Line 10 heart stained with anti GFP antibody which also detects GCaMP8.

Line 10 heart stained with anti GFP antibody which also detects GCaMP8, showing expression in cardiomyocytes.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Video showing GCaMP8 activity in heart, showing an increase in fluorescence with increased calcium during contraction.

 

 

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 αMHC-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 αMHC-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.

 

Comments are closed