B3

B3: acta2-RCaMP1.07 Strain Information Sheet

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 (PDF mouse profile sheet acta2-RCaMP1.07)

Strain Name:

Common Name:  acta2-RCaMP1.07

CHROMusTM designation and lines available: R24:B3:L2; R24:B3:L3

Jackson Stock number:  NA

Development:  The red fluorescent calcium indicator RCaMP1.07 (a gift from Dr. Masamichi Ohkura, Brain Science Institute, Saitama University) was inserted at the ATG site of the acta2 gene in the BAC RP23-370F21 (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
18 3 3

Description:  The genetically encoded calcium indicator RCaMP1.07 is expressed in smooth muscle cells in blood vessels, airways of the lung and gut.  Expression may be present in other smooth muscle containing tissues which have not been examined.  RCaMP1.07 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 smooth muscle tissues.

Phenotypic Data:

Figure 1: Native fluorescent images (RCaMP1.07) showing expression in frozen sections of A) heart and B) lung. v: vessel b: bronchiole

Figure 1: Native fluorescent images (RCaMP1.07) showing expression in frozen sections of A) heart and B) lung. v: vessel b: bronchiole

Figure 2: Native fluorescence (RCaMP1.07) in whole organs A) brain 2x B) brain 5x C) vessel 5x D) lung 2.7x

Figure 2: Native fluorescence (RCaMP1.07) in whole organs A) brain 2x B) brain 5x C) vessel 5x D) lung 2.7x

 

 

 

 

 

 

 

 

 

 

 

Figure 3: Calcium signaling in the small intestine.

 

Figure 4: Calcium signaling in the heart, primarily in the atrium and in the vessels.

 

 

Genotyping Protocol:

Primer B119 GCT TGT CTG TAA GCG GAT GCC
Primer B120 TGC TGC TGC CAC TCT AGT GAG AAA
Expected size of product: 602bp
Cycling conditions:

  1. 94oC 3 minutes
  2. Repeat 30 times
    1. 30 sec @ 94oC
    2. 30 sec @ 58oC
    3. 30 sec @ 72oC
  3. 72oC 5 minutes
  4. 10oC hold

PCR results:

B3 genotyping results

 

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 acta2-RCaMP1.07 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 acta2-RCaMP1.07 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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