Mutation of the <i>rsh</i> gene does not influence conditioning or place memory tested directly after training.

<p>Following a 30 s pre-test period (black bars), wild-type CS and <i>rsh¹</i> mutant flies were trained in two equal length periods for a total of either 6 or 20 min with 41°C (light gray bars). A 3 min memory was tested directly following in the post-test period (dark gray bars). The training, retention intervals, and testing patterns (both pre and post) are diagrammed for each panel, the time axis is not to scale. (<i>A</i>), Conditioning and memory tests were similar between the genotypes with 6 min of training (N = 331; pre-test: U = 12753.5, z = 1.07, P = 0.28; 1^st training period: U = 11877.0, z = 2.08, P = 0.04; 2^nd training period: U = 12888.5, z = 0.92, P = 0.36; post-test: U = 13237.0, z = 0.51, P = 0.61). (<i>B</i>) Conditioning and memory tests were also similar between the genotypes with 20 min of training (N = 232; pre-test: U = 6106.5, z = 1.22, P = 0.22; 1^st training period: U = 5740.5, z = 1.93, P = 0.06; 2^nd training period: U = 5802.0, z = −1.81,  = 0.07; post-test: U = 6463.0, z = −0.52, P = 0.60). (<i>C</i>) The <i>rsh</i> gene is necessary for normal short-term place memory. Flies were trained with intermittent training and then held for varying times (1 – 40 min) before being tested for memory with a short reminder training. The <i>rsh¹</i> flies had memory performance similar to wild-type CS levels with a 1 min delay between training and the memory test (N = 447, U = 24641.5, z = 0.24, P = 0.8). Significant differences were found at several time points following training (10 min: N = 295, U = 8637.0, z = .02, **  =  P<0.01; 20 min: N = 330, U = 10074.5, z = 3.95, ***  =  P<0.001; 30 min: N = 311, U = 10926.0, z = 1.45, P = 0.1; 40 min: N = 351, U = 12941.5, z = 2.48, **  =  P<0.01). The values are means and error bars represent s.e.m.</p

Control behaviors of wild-type CS and <i>rsh¹</i> mutant flies.

<p>MCH avoidance: ANOVA F(3,32)  = 1.07, P = 0.4; Oct avoidance: F(3,20)  = 1.3, P = 0.3; Sugar attractiveness: ANOVA F(3,44) = 0.75, P = 0.53; Activity: F(1,561) = 3.3, P = 0.07.</p

Mutation of the <i>rsh</i> gene reveals a major role in aversive olfactory memory (ARM) and is necessary for appetitive olfactory memory shortly after conditioning.

<p>Flies were either trained with odorants paired with electric shock or sugar reward. The training, cold-shock, retention intervals, and testing patterns (both pre and post) are diagrammed for each panel, the time axis is not to scale. (<i>A</i>) Olfactory memory tested three min after training is reduced in <i>rsh¹</i> flies compared to CS flies, although levels do not reach statistical significance (F(1,12)  = 3.5, P = 0.09). To reveal the <i>rsh</i> function in aversive olfactory memory, wild-type CS and <i>rsh¹</i> flies were trained with odorant / shock pairings, then after 2 hrs were given a cold-shock, memory was tested 1 hr later. Memory performance of <i>rsh¹</i> flies was significantly lower than wild-type CS flies with this procedure (F(1,10)  = 5.0, *  =  P = 0.04). (<i>B</i>) Appetitive olfactory short-term memory was tested at 3, 30, and 60 min after the odorant / sucrose training session. A <i>rsh¹</i> phenotype was evident at all tested time points after training (3 min: F(1,16)  = 29.2, ***  =  P<0.001; 30 min: F(1,14)  = 12.3, **  =  P<0.01; 60 min: F(1,14)  = 12.1, **  =  P<0.01). (<i>C</i>) The <i>rsh¹</i> appetitive short term olfactory memory phenotype is rescued with a transgenic copy of the wild-type version of the <i>rsh</i> gene (F(3,32)  = 13.0, P<0.0001; post-hoc tests: CS vs <i>rsh¹</i> ***  = P<0.001, <i>rsh¹</i> vs. <i>rsh¹</i>; hs-<i>rsh-1</i> *  = P<0.05, CS vs. <i>rsh¹</i>; hs-<i>rsh-1</i>, *  = P<0.05; <i>rsh¹</i> vs. CS; hs-<i>rsh-1</i> *  = P<0.05; CS vs. CS; hs-<i>rsh-1</i> *  = P<0.05). The values are means and error bars represent s.e.m.</p