dBACE overexpression in adult MB does not alter memory.

<p>Flies were fed with RU for 48 h before conditioning to induce <i>UAS-dBACE</i> transgene expression. STM assessed 2 h after one training session is not affected. The score of <i>MBSw</i>/<i>dBACE</i> flies is not different from that of the genetic control groups (<i>F</i>_(2,77) = 4.048, *<i>p</i> = 0.0214, <i>n</i> ≥ 24, Newman–Keuls <i>post-hoc</i>, <i>MBSw</i>/<i>dBACE</i> vs +/<i>dBACE p</i> > 0.05, <i>MBSw</i>/<i>dBACE</i> vs <i>MBSw</i>/+ <i>p</i> > 0.05). Bars, Mean ± SEM. PI, Performance index.</p

dBACE overexpression exacerbates the memory deficit caused by APPL partial loss-of-function.

<p>Unless indicated (A, w/o RU), flies were fed with RU for 48 h. (A) Flies were submitted to one cycle training and tested 2 h later. <i>Appl</i>^<i>d</i><i>/+;MBSw/+</i> flies show an STM deficit (<i>F</i>_(4,101) = 11.99, ***<i>p</i> < 0.0001, <i>n</i> ≥ 14, Newman-Keuls <i>post-hoc</i>, <i>Appl</i>^<i>d</i><i>/+;MBSw/+</i> vs <i>+</i> *<i>p</i> < 0.05), and <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dBACE</i> flies exhibit a STM score significantly lower than the genetic controls (Newman-Keuls <i>post-hoc</i>, <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dBACE</i> vs <i>Appl</i>^<i>d</i><i>/+;MBSw/+ ***p <</i> 0.001, <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dBACE</i> vs <i>+</i>/<i>dBACE ***p <</i> 0.001). <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dBACE</i> flies not fed with RU (w/o RU) display a STM score significantly different from flies of the same genotype fed with RU (Newman-Keuls <i>post-hoc</i>, *<i>p</i> < 0.05), and similar to <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>+</i> flies (Newman-Keuls <i>post-hoc</i>, <i>p</i> > 0.05). (B) Learning is not affected. To assess learning, flies were tested immediately after one cycle training (<i>F</i>_(2,27) = 0.8522, <i>p</i> = 0.4385, <i>n</i> ≥ 8). (C) Neither shock reactivity (<i>F</i>_(2,21) = 2.747, <i>p</i> = 0.0896, <i>n</i> ≥ 7) nor olfactory acuity (octanol, <i>F</i>_(2,46) = 0.3490, <i>p</i> = 0.7073, <i>n</i> ≥ 15; methylcyclohexanol, <i>F</i>_(2,52) = 2.959, <i>p</i> = 0.0610, <i>n</i> ≥ 17) is impaired. Bars, Mean ± SEM. PI, Performance index. (D, E) Analysis of <i>dBACE</i> and <i>Appl</i> transcription. Total RNA was extracted from <i>MBSw</i>/<i>dBACE</i>, <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/+ and <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dBACE</i> heads. Resulting cDNA was quantified using tubulin (Tub) expression as a reference. Results shown are ratios to the reference. (D) Quantification of <i>dBACE</i> mRNA level (<i>t</i> test, <i>p</i> = 0.8376, <i>n</i> = 4). (E) Quantification of <i>Appl</i> mRNA level (<i>t</i> test, <i>p</i> = 0.2330, <i>n</i> = 3). Bars, Mean ± SEM. ns, not significant.</p

dAβ expression alters memory only when associated with APPL partial loss-of-function.

<p>Unless indicated (B, w/o RU), flies were fed with RU for 48 h prior to either conditioning or mRNA extraction. (A, B) Flies were submitted to one cycle training and memory was tested 2 h later. (A) <i>MBSw</i>/<i>dAβ</i> flies exhibit a STM score similar to one of the genetic control groups (<i>F</i>_(2,53) = 8.421, ***<i>p</i> = 0.0007, <i>n</i> ≥ 17, Newman-Keuls <i>post-hoc</i>, <i>MBSw</i>/<i>dAβ</i> vs <i>+</i>/<i>dAβ p</i> > 0.05, <i>MBSw</i>/<i>dAβ</i> vs <i>MBSw</i>/<i>+</i> **<i>p</i> < 0.01). (B) <i>Appl</i>^<i>d</i><i>/+;MBSw/+</i> flies show an STM deficit (<i>F</i>_(4,144) = 12.41, ***<i>p</i> < 0.0001, <i>n</i> ≥ 26, Newman-Keuls <i>post-hoc</i>, <i>Appl</i>^<i>d</i><i>/+;MBSw/+</i> vs <i>+</i> **<i>p</i> < 0.01). <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dAβ</i> flies display a score significantly decreased compared to the controls (Newman-Keuls <i>post-hoc</i>, <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dAβ</i> vs <i>Appl</i>^<i>d</i><i>/+;MBSw/+ **p</i> < 0.01, <i>Appl</i>^<i>d</i>/+; <i>MBSw</i>/<i>dAβ</i> vs <i>+</i>/<i>dAβ ***p</i> < 0.001). <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dAβ</i> flies not fed with RU (w/o RU), exhibit scores significantly higher than flies of the same genotype fed with RU (Newman-Keuls <i>post hoc</i>, *<i>p</i> < 0.05), and similar to that of <i>Appl</i>^<i>d</i><i>/+;MBSw/+</i> control flies (Newman-Keuls <i>post hoc</i>, <i>p</i> > 0.05). (C) Learning is not affected. To assess learning, flies were submitted to one cycle training and tested immediately after (<i>F</i>_(2,30) = 1.684, <i>p</i> = 0.2038, <i>n</i> ≥ 9). (D) <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dAβ</i> flies exhibit wild-type shock reactivity (<i>F</i>_(2,34) = 0.7100, <i>p</i> = 0.4992, <i>n</i> ≥ 11), as well as wild-type olfactory acuity (octanol, <i>F</i>_(2,34) = 0.0507, <i>p</i> = 0.9506, <i>n</i> ≥ 11; methylcyclohexanol, <i>F</i>_(2,35) = 0.1433, <i>p</i> = 0.8670, <i>n</i> = 12). (E, F) Quantitative PCR analyses. RNA was extracted from <i>MBSw</i>/<i>dAβ</i><i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/+ and <i>Appl</i>^<i>d</i>/+;<i>MBSw</i>/<i>dAβ</i> fly heads. Resulting cDNA was quantified using tubulin (Tub) expression as a reference. Results shown are ratios to the reference. (E) Quantification of <i>dAβ</i> mRNA level. PCR reactions were conducted with primers specific of the <i>UAS-dAβ</i> construct (<i>t</i> test, <i>p</i> = 0.5788, <i>n</i> = 2). (F) Quantification of <i>Appl</i> mRNA level (<i>t</i> test, <i>p</i> = 0.1931, <i>n</i> = 2). Bars, Mean ± SEM. PI, Performance index. ns, not significant.</p