Source Paper
Stefano Tarantini, Marta Noa Valcarcel-Ares, Peter Toth, Andriy Yabluchanskiy, Zsuzsanna Tucsek et al.
Redox Biology • 2019
Adjustment of cerebral blood flow (CBF) to neuronal activity via neurovascular coupling (NVC) has an essential role in maintenance of healthy cognitive function. In aging increased oxidative stress and cerebromicrovascular endothelial dysfunction impair NVC, contributing to cognitive decline. There is increasing evidence showing that a decrease in NAD<sup>+</sup> availability with age plays a critical role in a range of age-related cellular impairments but its role in impaired NVC responses remains unexplored. The present study was designed to test the hypothesis that restoring NAD<sup>+</sup> concentration may exert beneficial effects on NVC responses in aging. To test this hypothesis 24-month-old C57BL/6 mice were treated with nicotinamide mononucleotide (NMN), a key NAD<sup>+</sup> intermediate, for 2 weeks. NVC was assessed by measuring CBF responses (laser Doppler flowmetry) evoked by contralateral whisker stimulation. We found that NVC responses were significantly impaired in aged mice. NMN supplementation rescued NVC responses by increasing endothelial NO-mediated vasodilation, which was associated with significantly improved spatial working memory and gait coordination. These findings are paralleled by the sirtuin-dependent protective effects of NMN on mitochondrial production of reactive oxygen species and mitochondrial bioenergetics in cultured cerebromicrovascular endothelial cells derived from aged animals. Thus, a decrease in NAD<sup>+</sup> availability contributes to age-related cerebromicrovascular dysfunction, exacerbating cognitive decline. The cerebromicrovascular protective effects of NMN highlight the preventive and therapeutic potential of NAD<sup>+</sup> intermediates as effective interventions in patients at risk for vascular cognitive impairment (VCI).
Objective: Assessment of learning and memory through discrimination between familiar and novel objects, sensitive to hippocampal and cortical microvascular function
This is a Novel Object Recognition Test protocol using mouse as the model organism. The procedure involves 6 procedural steps, 16 equipment items, 6 materials. Extracted from a 2019 paper published in Redox Biology.
Model and subjects
mouse • C57BL/6 • male • 24 months (aged cohort) • 20
Study window
~4.2 hours hands-on
Core workflow
Habituation phase • Acquisition (familiarization) phase • Delay period
Primary readouts
Key equipment and reagents
Verified items
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Animals explore empty open-field arena to acclimate to testing environment
Note: First phase of novel object recognition test
“During the habituation phase the animals explored the empty open-field arena for 5 min”
Mice explore two identical objects in the open-field arena
Note: Objects are made of washable odorless plastic, different shapes and colors but identical in size
“in the acquisition phase the mice explore two identical objects for 2 min”
Time interval between acquisition and trial phases
Note: Allows assessment of memory retention
“After a 4 h delay, a trial phase occurred”
Animals explore one familiar object and one novel object in the arena
Note: Exploration defined as directing nose at distance ≤2 cm to object and/or touching with nose. Sitting or climbing not considered exploration
“During this period animals explored the familiar object and a novel object for 2 min. Exploration of the objects was defined as directing the nose at a distance ≤2 cm to the object and/or touching it with the nose”
Clean arena and objects between trials to prevent olfactory cues
Note: Use 70% ethanol for cleaning
“The arena and the objects were cleaned with 70% ethanol between the trials to prevent the existence of olfactory cues”
Record and analyze exploration time using Ethovision software
Note: Calculate Recognition Index (RI) as measure of retention
“For data collection and analysis Ethovision software (Noldus Information Technology Inc., Leesburg, VA, USA) was used”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
Assessment of learning and memory through discrimination between familiar and novel objects, sensitive to hippocampal and cortical microvascular function
Objective
Assessment of learning and memory through discrimination between familiar and novel objects, sensitive to hippocampal and cortical microvascular function
Subjects
From papermouse • C57BL/6 • male • 24 months (aged cohort)
Sample count
From paper20
Cohort notes
From paperAged mice from National Institute on Aging colony at Charles River Laboratories
Habituation phase (5 minutes)
Acquisition (familiarization) phase (2 minutes)
Delay period (4 hours)
Trial phase (2 minutes)
Time spent exploring novel object relative to total exploration time
From paperA percent of time spent exploring the novel object relative to the total time spent exploring both objects was used as a measure of novel object recognition.
Artifact type
Longitudinal gait metrics and per-animal performance tables
Comparison focus
Compare recovery trajectory across post-injury timepoints and treatment conditions
Recognition Index (RI) = T novel / (T novel + T familiar)
From paperA percent of time spent exploring the novel object relative to the total time spent exploring both objects was used as a measure of novel object recognition.
Artifact type
Longitudinal gait metrics and per-animal performance tables
Comparison focus
Compare recovery trajectory across post-injury timepoints and treatment conditions
Discrimination between familiar and novel objects
From paperA percent of time spent exploring the novel object relative to the total time spent exploring both objects was used as a measure of novel object recognition.
Artifact type
Longitudinal gait metrics and per-animal performance tables
Comparison focus
Compare recovery trajectory across post-injury timepoints and treatment conditions
Learning and memory performance
From paperA percent of time spent exploring the novel object relative to the total time spent exploring both objects was used as a measure of novel object recognition.
Artifact type
Longitudinal gait metrics and per-animal performance tables
Comparison focus
Compare recovery trajectory across post-injury timepoints and treatment conditions
Time spent exploring novel object relative to total exploration time
From paperRaw 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
Final reported form
Group comparisons of gait indices, stride metrics, or recovery curves
Recognition Index (RI) = T novel / (T novel + T familiar)
From paperRaw 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
Final reported form
Group comparisons of gait indices, stride metrics, or recovery curves
Discrimination between familiar and novel objects
From paperRaw 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
Final reported form
Group comparisons of gait indices, stride metrics, or recovery curves
Learning and memory performance
From paperRaw 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
Final reported form
Group comparisons of gait indices, stride metrics, or recovery curves
Acquisition
Capture run-level gait data for each animal and preserve the timepoint or treatment labeling.
Preprocessing / cleaning
A percent of time spent exploring the novel object relative to the total time spent exploring both objects was used as a measure of novel object recognition.
Scoring or quantification
Quantify the primary readouts for this experiment: Time spent exploring novel object relative to total exploration time; Recognition Index (RI) = T novel / (T novel + T familiar); Discrimination between familiar and novel objects; Learning and memory performance.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Time spent exploring novel object relative to total exploration time, Recognition Index (RI) = T novel / (T novel + T familiar), Discrimination between familiar and novel objects, Learning and memory performance.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Stefano Tarantini et al. (2019). Nicotinamide mononucleotide (NMN) supplementation rescues cerebromicrovascular endothelial function and neurovascular coupling responses and improves cognitive function in aged mice. Redox Biology
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Noldus Information Technology Inc.
Columbus Instruments
Chatillon Ametek Force Measurement
Noldus Information Technology Inc.
Kent Scientific Co • MousVent G500
Kent Scientific Co
Living Systems Instrumentations
Leica Microsystems
Transonic Systems Inc.
Noldus Information Technology Inc.
Noldus Information Technology Inc.
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Current status surfaces were computed from experiment data updated Mar 14, 2026.
Source access
Jump back into the original paper or the methods evidence section when you need exact wording, exclusions, or method-specific caveats.
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Steps
6
Evidence Quotes
28
Protocol Items
22
Linked Products
8
Canonical Sync
Pending
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Computed from the current experiment record updated Mar 14, 2026.
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Steps
6
Evidence
28
Specific Products
8/8
Canonical Sync
Pending
What this score means
The verification score reflects evidence coverage, subject detail, paper provenance, step depth, and whether linked products resolve to specific item pages instead of generic searches.
Computed from the current experiment record updated Mar 14, 2026.
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