Rotarod Trials and Performance Analysis
Explore the detailed analysis of Rotarod trials conducted on Q58 mice, including protocols, results, and comparisons with published data. Discover insights into motor phenotype verification and performance differences between genotypes and genders.
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Presentation Transcript
Q58 Rotarod Trials 2/13/15 - Matthew Schneider
Q58 Rotarod Trials Received the Q58 mice in an FVB background from the Bezprozvanny lab last year Bred mice for a rotarod group to verify motor phenotype in our hands We had not previously seen a rotarod phenotype in this particular background
Q58 Rotarod Protocol 28 Mice 14 Wt and 14 Tg 7 males and 7 females for each genotype Tested at 4 months, 6 months, and 8 months 5 day rotarod protocol Monday mice are handled Tuesday training day Wednesday, Thursday, Friday Accelerating rotarod from 0 to 40 rpm in 360 seconds; 3 trials each day No weight differences between Tg and Wt mice at any of the time points
Q58 Rotarod Results All Mice 350 300 250 Time to fall (s) 200 150 100 50 0 4 Mo. 6 Mo. 8 Mo. Wt Tg Females Males 350 350 300 300 250 Time to fall (s) 250 Time to fall (s) Data presented using SEM for error bars 200 200 150 150 100 100 50 50 0 0 4 Mo. 6 Mo. 8 Mo. 4 Mo. 6 Mo. 8 Mo. Wt Tg Wt Tg
Comparison of the rotarod performance of Q58 mice in our lab to Bezprozvanny s published data ** p < 0.01 SCA2-58Q mice males bred with B6D2 Company females SCA2-58Q (N6 FVB) males bred with FVB Company females Only females used Rotarod Protocol: Mice are screened at 5 rpm, if they fall off before 5 minutes they aren t used Tested for 4 days and results analyzed from day 4 (3 trials/day) 0 40 rpm in 200 seconds
Comparison of the rotarod performance of Q58 mice in our lab to Bezprozvanny s published data SCA2-58Q (N6+ FVB) males bred with FVB Company females Animals were 9 months old at experiment start Rotarod Protocol Pre-drug feeding: beamwalk, then rotarod, same testing as the first two Post-drug feeding: Day 1 beamwalk training Day 2 beamwalk testing Day 3-5 Rest Day 6 rotarod training (3 trials/day) Day 7 rotarod testing (3 trials/day)
ITPR1 ASO Work From a pool of ASOs we received the two most promising based on in-vitro work Injected the ASOs into mice to determine possible ITPR1 knockdown
Itpr1 ASOs lower Itpr1 mRNA abundance ITPR1 P<0.001 for both mAtxn2 Iba1 4.5 1.4 2.5 P<0.001 4 1.2 2 3.5 1 mAtxn2 / Actin Itpr1 / Actin 3 Iba1 / Actin 1.5 2.5 0.8 2 1 0.6 1.5 1 0.4 0.5 0.5 0.2 0 0 ASO 178 ASO 195 Saline 0 ASO 178 ASO 195 Saline ASO 178ASO 195 Saline N=3 males and 3 females per treatment All wildtype mice Mice were 5 months old 250 g ASO treated for 7 days qPCR results shown
Q72 BAC Mouse Rotarod Behavioral Testing
Q72 BAC Mice Tested to verify rotarod phenotype with new protocol and different mouse background 28 Mice 14 Wt and 14 Tg 7 males and 7 females for each genotype Tested at 4 months, 6 months, and 8 months 5 day rotarod protocol Monday mice are handled Tuesday training day Wednesday, Thursday, Friday Accelerating rotarod from 0 to 40 rpm in 360 seconds; 3 trials each day
Q72 BAC Differences Significant weight differences between Wt and Tg Mice Tg mice were visibly more active ( bouncy ) All Mice * 30 25 20 Weight (g) 15 10 5 0 Female Tg Wt Male 30 * 35 25 * 30 Weight (g) 20 25 Weight (g) 15 20 10 15 5 10 0 5 Tg - F Wt - F 0 Tg - M Wt - M
Rotarod Performance Results Day 2 Day 1 * * 250 200 180 200 160 140 Time to fall (s) Time to fall (s) 150 120 100 100 80 60 50 40 20 0 0 Wt Tg Wt Tg
Rotarod Performance Results Cont. Day 3 250 * Avg. Day 3 200 300 time to fall (s) 150 250 100 200 Time to fall (s) 50 150 0 Wt Tg 100 50 0 Wt Wt Wt Wt Wt Wt Wt Tg Tg Tg Tg Tg Tg Tg Wt Wt Wt Wt Wt Wt Wt Tg Tg Tg Tg Tg Tg Tg F F F F F F F F F F F F F F M M M M M M M M M M M M M M
