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View Abstract
Establishing standard operating procedures (SOPs) as tools for the analysis of behavioral phenotypes is fundamental to mouse functional genomics. It is essential that the tests designed provide reliable measures of the process under investigation but most importantly that these are reproducible across both time and laboratories. For this reason, we devised and tested a set of SOPs to investigate mouse behavior. Five research centers were involved across France, Germany, Italy, and the UK in this study, as part of the EUMORPHIA program. All the procedures underwent a cross-validation experimental study to investigate the robustness of the designed protocols. Four inbred reference strains (C57BL/6J, C3HeB/FeJ, BALB/cByJ, 129S2/SvPas), reflecting their use as common background strains in mutagenesis programs, were analyzed to validate these tests. We demonstrate that the operating procedures employed, which includes open field, SHIRPA, grip-strength, rotarod, Y-maze, prepulse inhibition of acoustic startle response, and tail flick tests, generated reproducible results between laboratories for a number of the test output parameters. However, we also identified several uncontrolled variables that constitute confounding factors in behavioral phenotyping. The EUMORPHIA SOPs described here are an important start-point for the ongoing development of increasingly robust phenotyping platforms and their application in large-scale, multicentre mouse phenotyping programs.
Protocol Steps
1
Establish Standard Operating Procedures
Devise and test a set of SOPs to investigate mouse behavior across five research centers in France, Germany, Italy, and the UK as part of the EUMORPHIA program
Not specifiedNot specified
Note: Cross-validation experimental study to investigate robustness of designed protocols
View evidence from paper
“we devised and tested a set of SOPs to investigate mouse behavior. Five research centers were involved across France, Germany, Italy, and the UK”
2
Conduct SHIRPA Test
Perform SHIRPA (Standardized SHIRPA and Research Profile Assessment) as part of the test battery for phenotypic assessment
Not specifiedNot specified
Note: SHIRPA is one of the operating procedures employed in the test battery
View evidence from paper
“the operating procedures employed, which includes open field, SHIRPA, grip-strength, rotarod, Y-maze, prepulse inhibition of acoustic startle response, and tail flick tests”
3
Conduct Additional Behavioral Tests
Perform complementary tests including open field, grip-strength, rotarod, Y-maze, prepulse inhibition of acoustic startle response, and tail flick tests
Note: All tests conducted as part of standardized test battery
View evidence from paper
“the operating procedures employed, which includes open field, SHIRPA, grip-strength, rotarod, Y-maze, prepulse inhibition of acoustic startle response, and tail flick tests”
4
Analyze Four Inbred Reference Strains
Test C57BL/6J, C3HeB/FeJ, BALB/cByJ, and 129S2/SvPas strains to validate the test procedures
Not specifiedNot specified
Note: Four inbred reference strains selected as they reflect common background strains in mutagenesis programs
View evidence from paper
“Four inbred reference strains (C57BL/6J, C3HeB/FeJ, BALB/cByJ, 129S2/SvPas), reflecting their use as common background strains in mutagenesis programs, were analyzed”
5
Cross-Laboratory Validation
Validate reproducibility of test results across multiple laboratories to identify robust output parameters and confounding variables
Not specifiedNot specified
Note: Study generated reproducible results between laboratories for a number of test output parameters but identified several uncontrolled variables
View evidence from paper
“generated reproducible results between laboratories for a number of the test output parameters. However, we also identified several uncontrolled variables that constitute confounding factors”
Subjects / Specimens
Species
mouse
Strain
Four inbred reference strains: C57BL/6J, C3HeB/FeJ, BALB/cByJ, 129S2/SvPas
Age
Not specified
Sex
unknown
Weight
Not specified
Four inbred reference strains reflecting their use as common background strains in mutagenesis programs
