Optimization of a contextual conditioning protocol for rats using combined measurements of startle amplitude and freezing: the effects of shock intensity and different types of conditioning.

Contextual conditioning in rats is typically quantified using startle amplitude or freezing time. Our goal was to create a robust contextual conditioning protocol combining both startle amplitude and freezing time as measures of contextual anxiety. Comparison of 0.8 mA - 250 ms shocks with an established shock configuration (0.3 mA - 1 s) favoured the first parameters. Subsequently, we systematically investigated the effect of shock intensity (0.6 mA, 0.8 mA or 1.0 mA) and concurrently compared two different contextual conditioning procedures (shocks alone versus unpaired shock-tone presentations). In future experiments, this second type of contextual conditioning may form the optimal contrasting condition for a cued fear conditioning group, trained with explicit cue-shock pairings. The 0.8 mA shocks produced significant contextual freezing and startle potentiation, whereas the 0.6 mA and 1.0 mA shocks only led to a significant increase of freezing time. We found no major differences between the two types of conditioning, implying that these procedures might be equivalent. In conclusion, training with ten 0.8 mA - 250 ms shocks produced reliable contextual conditioning as measured with both startle amplitude and freezing time.