TABLE 1.

Variables for consideration, key to behavioral outcomes

Guidance	Addresses	Recommend	References
Sample size blinding randomization	Power false reporting	N = 15–20 per genotype/treatment/sex	50, 51, 52
Age and sex	Age‐dependent effects	One sex cannot be tested; genders cannot be combined with powered Ns and statistics showing no difference; Animals of wildly different ages cannot be compared	53, 54, 55
	Sex‐specific effects
Breeding schema	Developmental environment	WT × WT and KO × KO is unacceptable; a KO's maternal behavior is likely different than WT; thus, het by het breeding recommended	51, 52, 55, 56, 57, 58, 59
Housing conditions	Developmental environment	Housed within genotype/treatment or mixed genotype/treatment; house isolated or grouped	51, 52, 58, 60
Background strain	Background genetics	Consistent reporting/congenic when possible	55, 61, 62
Order of testing	Test‐re test influencing on behavior	Non stressful to stressful	63, 64, 65
Littermates or litter effects	Developmental environment consistency	Reporting and accounting for appropriate control subjects accounting for maternal care; litter size	66, 67, 68, 69
Task validation	Tests must be validated to measure what you are assessing.	Examples include a benzodiazepine will change behavior in an anxiety test and/or a sedative lower motor activity	57
Reproducibility	Chance findings/type 1 error	For rigorous, reliable behavioral outcome measures	59, 70