Treatment with the mitochondrial-targeted antioxidant peptide SS -31 rescues neurovascular coupling responses and cerebrovascular endothelial function and improves cognition in aged mice methods
Aim. Evidence-backed execution summary for Treatment with the mitochondrial-targeted antioxidant peptide SS -31 rescues neurovascular coupling responses and cerebrovascular endothelial function and improves cognition in aged mice methods from Treatment with the mitochondrial-targeted antioxidant peptide SS -31 rescues neurovascular coupling responses and cerebrovascular endothelial function and improves cognition in aged mice.
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mouse
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Measurement of neurovascular coupling responses
reagent used in the protocol.
- Use
- After behavioral testing, mice in each group were anesthetized with isoflurane (4% induction and 1% maintenance), endotracheally intubated, and ventilated. Neurovascular coupling responses were assessed by measuring changes in cerebral blood flow above the left barrel cortex using laser speckle contrast imaging in r...
Assessment of endothelial NO-mediated vasodilation in isolated cerebral vessels
reagent used in the protocol.
- Use
- To assess the specific effect of SS-31 treatment on endothelial NO mediation, isolated segments of the middle cerebral arteries were cannulated and pressurized, as reported (Springo et al.,; Toth et al., ). Dilations to acetylcholine (10 -8 -10 -5 M) and ATP (10 -7 ...
Restoration of cerebromicrovascular function is associated with improved cognitive function in aged mice treated with...
Previous studies show that selective pharmacologically induced neurovascular uncoupling is associated with cognitive impairment (Tarantini et al., ). To determine how restoration of cerebromicrovascular function impacts cognitive function in aged mice, animals were tested in the radial-arm water maze (Fig...
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- Previous studies show that selective pharmacologically induced neurovascular uncoupling is associated with cognitive impairment (Tarantini et al., ). To determine how restoration of cerebromicrovascular function impacts cognitive function in aged mice, animals were tested in the radial-arm water maze (Fig...
Restoration of cerebromicrovascular function is associated with improved cognitive function in aged mice treated with...
In SS -31-treated aged mice, rescue of neurovascular coupling responses associates with improved radial-arm water maze ( RAWM ) performance. Young (3 months old), aged (24 months old), and SS -31-treated aged mice were tested in the RAWM. Panel a: Heatmap representing th...
- Use
- In SS -31-treated aged mice, rescue of neurovascular coupling responses associates with improved radial-arm water maze ( RAWM ) performance. Young (3 months old), aged (24 months old), and SS -31-treated aged mice were tested in the RAWM. Panel a: Heatmap representing th...
Rotarod, motor skill learning
Motor coordination was assessed using the rotarod as described (Tarantini et al., ). Analysis of day-to-day changes in performance on the accelerating rotarod test was used to evaluate the motor skill learning. The grid hanging test was also used to evaluate balance and motor function. A grip streng...
- Use
- Motor coordination was assessed using the rotarod as described (Tarantini et al., ). Analysis of day-to-day changes in performance on the accelerating rotarod test was used to evaluate the motor skill learning. The grid hanging test was also used to evaluate balance and motor function. A grip streng...
Restoration of cerebromicrovascular function is associated with improved cognitive function in aged mice treated with...
Successful escape rate from the maze was assessed by measuring the percent of animals that could find the hidden platform within the 60 s allowed for each trial. During acquisition, mice from all groups showed an increase in successful escape rate consistent with the learning of the task. Young mice exhibited s...
- Use
- Successful escape rate from the maze was assessed by measuring the percent of animals that could find the hidden platform within the 60 s allowed for each trial. During acquisition, mice from all groups showed an increase in successful escape rate consistent with the learning of the task. Young mice exhibited s...
Restoration of cerebromicrovascular function is associated with improved cognitive function in aged mice treated with...
We also compared path length (i.e., the distance that the mouse swam between maze entry and successful escape through the hidden platform) and escape latency (i.e., the time elapsed between entry and successful escape). During acquisition, mice from all groups displayed shorter path length (Figure e) and lower...
- Use
- We also compared path length (i.e., the distance that the mouse swam between maze entry and successful escape through the hidden platform) and escape latency (i.e., the time elapsed between entry and successful escape). During acquisition, mice from all groups displayed shorter path length (Figure e) and lower...
Treatment with SS-31 restores motor learning function in aged mice
Motor skill learning was assessed using a modified rotarod test. During a 4-day-long experimental period, young mice displayed a steeper motor skill learning curve compared to aged control mice. After the first day, aged control animals consistently fell off the accelerating rotarod earlier compared to y...
- Use
- Motor skill learning was assessed using a modified rotarod test. During a 4-day-long experimental period, young mice displayed a steeper motor skill learning curve compared to aged control mice. After the first day, aged control animals consistently fell off the accelerating rotarod earlier compared to y...
Analysis of gait function
To determine how aging and SS-31 treatment affect gait coordination, we tested the animals using an automated computer-assisted method (CatWalk; Noldus Information Technology Inc. Leesburg, VA, USA) as described (Tarantini et al.,; Toth, Tarantini, Springo et al., ).
- Use
- To determine how aging and SS-31 treatment affect gait coordination, we tested the animals using an automated computer-assisted method (CatWalk; Noldus Information Technology Inc. Leesburg, VA, USA) as described (Tarantini et al.,; Toth, Tarantini, Springo et al., ).
Treatment with SS-31 restores motor learning function in aged mice
SS -31 treatment in aged mice improves motor skill learning but not gait coordination. a) Motor skill learning on the accelerating rotarod. Daily changes in mean latencies to fall for young (3 months old), aged (24 months old) and SS -31-treated aged mice are shown. Data are mean ...
- Use
- SS -31 treatment in aged mice improves motor skill learning but not gait coordination. a) Motor skill learning on the accelerating rotarod. Daily changes in mean latencies to fall for young (3 months old), aged (24 months old) and SS -31-treated aged mice are shown. Data are mean ...
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Rotarod, motor skill learning
Motor coordination was assessed using the rotarod as described (Tarantini et al., ). Analysis of day-to-day changes in performance on the accelerating rotarod test was used to evaluate the motor skill learning. The grid hanging test was also used to evaluate balance and motor function. A grip strength test was used to measure the maximal muscle strength of forelimbs of the mice.
Restoration of cerebromicrovascular function is associated with improved cognitive function in aged mice treated with...
To analyze working memory function (short-term memory that is involved in immediate conscious perception), we examined re-entries into incorrect arms (without hidden platform) that were previously attempted for escape. We found that working memory function was impaired in aged mice as compared to young controls (Figure c). Aged mice that were treated with SS-31 showed significant restoration of working memory to levels comparable to young animals (Figure c). SS-31 treatment thus resulted in a marked improvement in working memory while there was no clear effect of SS-31 on learning.
EXPERIMENTAL PROCEDURES
Young (3 months, n = 30) and aged (24 months, n = 40) male C57BL/6 mice were used. Mice in the aged cohort were assigned to two groups ( n = 20 each group). One group of the aged mice was injected daily with SS-31 (elamipretide; also known as MTP-131 or Bendavia, 10 mg Kg -1 day -1, for 10 days). This dosage of SS-31 has been shown to exert potent cardioprotective effects (Dai et al., ). All procedures were approved by the Institutional Animal Use and Care Committees of the University of Oklahoma Health Sciences Center.
Statistical analysis
Data were analyzed by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. Principal component analysis, followed by MANOVA and the PCA-biplot approach were used to analyze the gait data. A p value less than.05 was considered statistically significant. Data are expressed as mean ± SEM.
Quantitative real-time RT-PCR
A quantitative real-time RT-PCR technique was used to analyze mRNA expression in cortical samples using validated TaqMan probes (Applied Biosystems) and a Strategen MX3000 platform, as previously reported (Toth, Tarantini, Ashpole et al., ).
Measurement outputs
What raw and processed outputs should exist?
Treatment with SS-31 did not alter the expression of Nos1 and Nos3 in the aged mouse brain (Figure h). The expression of arginase 1 was increased in aged mice and te...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
Previous studies show that selective pharmacologically induced neurovascular uncoupling is associated with cognitive impairment (Tarantini et al., ). To determine how resto...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
In SS -31-treated aged mice, rescue of neurovascular coupling responses associates with improved radial-arm water maze ( RAWM ) performance. Young (3 mont...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
Motor coordination was assessed using the rotarod as described (Tarantini et al., ). Analysis of day-to-day changes in performance on the accelerating rotarod t...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
Analysis plan
How should the outputs become interpretable results?
Acquisition
Capture run-level gait data for each animal and preserve the timepoint or treatment labeling.
inferred from protocolPreprocessing / cleaning
To further ascertain the endothelial protective effects of SS-31, endothelium-dependent vasodilator responses were tested in isolated, cannulated branches of the MCA.
from paperScoring or quantification
Quantify the primary readouts for this experiment: Treatment with SS-31 did not alter the expression of Nos1 and Nos3 in the aged mouse brain (Figure h). The expression of arginase 1 was increased in aged mice and te...; Previous studies show that selective pharmacologically induced neurovascular uncoupling is associated with cognitive impairment (Tarantini et al., ). To determine how resto...; In SS -31-treated aged mice, rescue of neurovascular coupling responses associates with improved radial-arm water maze ( RAWM ) performance. Young (3 mont...; Motor coordination was assessed using the rotarod as described (Tarantini et al., ). Analysis of day-to-day changes in performance on the accelerating rotarod t....
from paperStatistical comparison
To further ascertain the endothelial protective effects of SS-31, endothelium-dependent vasodilator responses were tested in isolated, cannulated branches of the MCA...; To substantiate the endothelial protective effects of SS-31 in vitro, we assessed the effects of SS-31 on cellular mtROS production in cultured CMVECs derived f...; In SS -31-treated aged mice, rescue of neurovascular coupling responses associates with improved radial-arm water maze ( RAWM ) performance. Young (3 mont...; To analyze working memory function (short-term memory that is involved in immediate conscious perception), we examined re-entries into incorrect arms (without hidden...
from paperReporting output
Report representative outputs alongside summary comparisons for Treatment with SS-31 did not alter the expression of Nos1 and Nos3 in the aged mouse brain (Figure h). The expression of arginase 1 was increased in aged mice and te..., Previous studies show that selective pharmacologically induced neurovascular uncoupling is associated with cognitive impairment (Tarantini et al., ). To determine how resto..., In SS -31-treated aged mice, rescue of neurovascular coupling responses associates with improved radial-arm water maze ( RAWM ) performance. Young (3 mont..., Motor coordination was assessed using the rotarod as described (Tarantini et al., ). Analysis of day-to-day changes in performance on the accelerating rotarod t....
inferred from protocolStructured statistical methods
To further ascertain the endothelial protective effects of SS-31, endothelium-dependent vasodilator responses were tested in isolated, cannulated branches of the MCA...; To substantiate the endothelial protective effects of SS-31 in vitro, we assessed the effects of SS-31 on cellular mtROS production in cultured CMVECs derived f...; In SS -31-treated aged mice, rescue of neurovascular coupling responses associates with improved radial-arm water maze ( RAWM ) performance. Young (3 mont...; To analyze working memory function (short-term memory that is involved in immediate conscious perception), we examined re-entries into incorrect arms (without hidden...
source structuredSource and audit
What supports the facts on this page?
Evidence quotes (5)
Motor coordination was assessed using the rotarod as described (Tarantini et al., ). Analysis of day-to-day changes in performance on the accelerating rotarod test was used to evaluate the motor skill learning. The grid hanging test was also used to evaluate balance and motor function. A grip strength test was used to measure the maximal muscle strength of forelimbs of the mice.
To analyze working memory function (short-term memory that is involved in immediate conscious perception), we examined re-entries into incorrect arms (without hidden platform) that were previously attempted for escape. We found that working memory function was impaired in aged mice as compared to young controls (Figure c). Aged mice that were treated with SS-31 showed significant restoration of working memory to levels comparable to young animals (Figure c). SS-31 treatment thus resulted in a marked improvement in working memory while there was no clear effect of SS-31 on learning.
Young (3 months, n = 30) and aged (24 months, n = 40) male C57BL/6 mice were used. Mice in the aged cohort were assigned to two groups ( n = 20 each group). One group of the aged mice was injected daily with SS-31 (elamipretide; also known as MTP-131 or Bendavia, 10 mg Kg -1 day -1, for 10 days). This dosage of SS-31 has been shown to exert potent cardioprotective effects (Dai et al., ). All procedures were approved by the Institutional Animal Use and Care Committees of the University of Oklahoma Health Sciences Center.
Data were analyzed by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. Principal component analysis, followed by MANOVA and the PCA-biplot approach were used to analyze the gait data. A p value less than.05 was considered statistically significant. Data are expressed as mean ± SEM.
A quantitative real-time RT-PCR technique was used to analyze mRNA expression in cortical samples using validated TaqMan probes (Applied Biosystems) and a Strategen MX3000 platform, as previously reported (Toth, Tarantini, Ashpole et al., ).
Machine-readable layer
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