Cognitive, learning and memory domain

Novel object recognition – This test, by evaluating the differences in the exploration time of novel and familiar objects, provides a measure for cognition, especially for recognition and episodic memory. The animal is put inside a transparent plexiglass box 50cm all dimensions, with two similar “familiar” objects for a habituation stage of 10 minutes. After an hour the animal is re-entered to the arena for a 4 minute test session, now with one “familiar” and one different “novel” object. Duration of time spent near each object, and the frequency and number of approaches are tracked.

[This test is applicable to rats and mice]

References
Lotan A, Lifschytz T, Wolf G, Keller S, Ben-Ari H, Tatarsky P, Pillar N, Oved K, Sharabany J, Merzel TK, Matsumoto T, Yamawaki Y, Mernick B, Avidan E, Yamawaki S, Weller A, Shomron N, Lerer B (2018). Differential effects of chronic stress in young-adult and old female mice: cognitive-behavioral manifestations and neurobiological correlates. 23(6) : 1432-1445.

Benhamron S, Keren Nitzan K, Valitsky M, Karussis D, Kassis I, Rosenmann H (2020). Amelioration of cognitive deficits and brain pathology by CSF exchange therapy with artificial CSF enriched with mesenchymal stem cell secretions in Alzheimer’s disease-mice. J Alzheimers Dis. 76(1).

Radial arm water maze – Evaluates working memory, spatial learning and cognitive ability. The apparatus consists of 6 arms, 30 cm in length converging on a central 40X40 cm pool, filled with water to cover a plexiglass platform. On day 1 of the protocol the animal is trained to locate the platform (15 trials of up to 1 minute, alternating between hidden and visible platforms). On day 2 (test) all 15 trials are with the platform hidden. Entry into the wrong arm or non-entry into the correct arm after 15 seconds are designated as errors. Time taken for each animal to rech the platform in each trial is designated as latency to platform. Number of errors for each animal and latencies to platform in each trial is tracked and scored.

[This test is applicable to rats and mice]

References

Nitzan K, Benhamron S, Valitsky M, Kesner EE, Lichtenstein M, Ben-Zvi A, Ella E, Segalstein Y, Saada A, Lorberboum-Galski H, Rosenmann H (2019). Mitochondrial Transfer Ameliorates Cognitive Deficits, Neuronal Loss, and Gliosis in Alzheimer’s Disease Mice. J Alzheimers Dis. 72(2):587-601

Ben-Ari H, Lifschytz T, Wolf G, Rigbi A, Blumenfeld-Katzir T, Merzel TK, Koroukhov N, Lotan A, Lerer B (2019). White matter lesions, cerebral inflammation and cognitive function in a mouse model of cerebral hypoperfusion. Brain Research 1711: 193-201.

T-maze – This test was used for assessing the spatial short-term memory and alternation behavior, that is, determining the mouse’s ability to recognize and differentiate between a new unknown and a familiar compartment The T-shaped maze is made of plastic with 2 arms 45 cm in length that extendes at a right angle from a 57-cm-long alley. The arms have a width of 10 cm and are surrounded by 10-cm-high walls. The test consists of 2 trials with an interval of 1 h (or alternatively of 24h), during which time the animals are put back to their home cages. During an 8-minute acquisition trial, one of the short arms is closed. In a 3-minute retention trial, mice have access to both arms and to the alley. The number of entries into the unfamiliar arm and the time spent in the unfamiliar arm are recorded. Mice normally tend to enter more times and spend more time in the new unknown arm than in the familiar one or in the alley.[This test is applicable to rats and mice]

References

Zueger M, Urani A, Chourbaji S, Zacher C, RocheM , Harkin A, Gass P (2005). Olfactory bulbectomy in mice induces alterations in exploratory behavior Neurosci. Lett., 374 (2) 142-146

Benhamron S, Rozenstein-Tsalkovich L, Nitzan K, Abramsky O, Rosenmann H (2018) . Phos-tau peptide immunization of amyloid-tg-mice reduced non-mutant phos-tau pathology, improved cognition and reduced amyloid plaques. Exp Neurol. 303:48-58.

Y-maze – This test evaluates short-term memory. The Y-maze is a three-arm maze with all arms at equal angles, 30 cm in length and 5 cm in width with walls 12 cm high. Mice are initially placed in the middle, and the sequence of arm entries are recorded manually for each mouse over a 8-minute period. The triads with all three arms represented (i.e., ABC, CAB, or BCA but not ABB) – are considered ‘correct triads’. [This test is applicable to rats and mice]

References

He C, Tsipis CP, LaManna JC, Xu K (2017). Environmental Enrichment Induces Increased Cerebral Capillary Density and Improved Cognitive Function in Mice. Adv Exp Med Biol 977, 175–181.

Eight arm maze – Evaluates working memory, spatial learning and cognitive ability. Using water deprivation (by limiting water consumption overnight) and a reward of 50 mcl water at end of each arm, each animal is introduced into the maze until either completing visiting all eight arms or 25 trial (whichever comes first). The lower number of arm entries needed to complete visit to all arms- the better the cognitive score. This procedure is repeated for five consecutive days.

References
Rosenmann H, Grigoriadis N, Eldar-Levy H, Avital A, Rozenstein L, Touloumi O, Behar L, Ben-Hur T, Avraham Y, Berry E, Segal M, Ginzburg I, Abramsky O.(2008) A novel transgenic mouse expressing double mutant tau driven by its natural promoter exhibits tauopathy characteristics. Eur Neurol 212(1): 71-84

Morris water maze – This test evaluate cognitive ability, especially short term (cortical) and longer term (hippocampal) working memory and spatial learning. The test apparatus is a round metal pool, filled with an opaque water to a height that covers a plexiglass platform 16X16 cm located 20 cm from the pool edge. In acquisition trials (4 consecutive for each animal, repeated for 5 days), the time required for the animal to find the platform is tracked and recorded. Reversal trials consist of time taken to find the platform relocated to the opposite side of pool. Probe trials consist of trials in clear water, without the platform, in which the time spent by the animal in each of four quadrants of the pool is tracked and recorded.

[This test is applicable to rats (pool diameter 140cm, height 60 cm) and mice (pool diameter 120 cm, height 40 cm)]

References

Mutlu O, Ulak G, Celikyurt IK, Tanyeri P, Akar FY, Erden F (2011) Effects of olanzapine and clozapine on memory acquisition, consolidation and retrieval in mice using the elevated plus maze test. Neurosci Lett 501(3): 143-147

Cued and context fear conditioning – This test aims to evaluate fear memory- contextual (hippocampus mediated) and cued (amygdale mediated). The test device consists of two compartments and has a grid floor capable of generating low electrical current for a given time period. The animal is put inside the device and receives a 2 second, 0.5 mA shock coupled to a tone for two repeats. After 48 hours, the freezing time of the animal after exposure to either the acoustic tone (cued fear conditioning) or the visual context (context fear conditioning) is measured out of a total 5 minute exposure.

[This test is applicable to rats and mice]

References
Lotan A, Lifschytz T, Mernick B, Lory O, Levi E, Ben-Shimol E, Goelman G, Lerer B (2017). Alterations in the expression of a neurodevelopmental gene exert long-lasting effects on cognitive-emotional phenotypes and functional brain networks: translational evidence from the stress-resilient Ahi1 knockout mouse. Molecular Psychiatry 22(6): 884-899.

Wolf G, Lifschytz T, Ben-Ari H, Tatarskyy P, Merzel TK, Lotan A, Lerer B (2018). Effect of chronic unpredictable stress on mice with developmental under-expression of the Ahi1 gene: behavioral manifestations and neurobiological correlates. Translational Psychiatry 8(1): doi: 10.1038/s41398-018-0171-1.

Attentional set shifting– This test aims to evaluate animal ability of the animal to learn rules associated with finding food, and its cognitive flexibility in re-learning as these rules are changed. Animals are first acclimated to a plexiglass chamber. Thereafter, two bowls are presented with different odors/mediums, one of them baited. Trials consist of the animal able to discern which of the cups contains food using simple discrimination (odor and medium), compound discrimination rules. Thereafter, the animals are required to adjust to changes in the location of the food (from one bowl to the other) according to intra-dimensional (same dimension- odor or medium change), and extra-dimensional (reward for medium instead of odor and vice versa) rules. For each session, number of correct choices and errors are recorded.

[This test is applicable to rats and mice]

References
Young JW, Powell SB, Geyer MA, Jeste DV, Risbrough VB (2010) The mouse attentional-set-shifting task: A method for assaying successful cognitive aging? Cogn Affect Behav Neurosci 10(2):243-251

Open field habituation – This test evaluates long term non-associative, non-aversive spatial learning by measuring the decrease in the exploratory activity of the animal in a test session carried out 24 h after the first exploration session (delta of 2nd session – 1st session). Animals are exposed to a novel environment by placing them in a 40 cm × 50 cm × 60 cm open field box. The distance walked is measured for a 5 min period (EthoVision). Twenty-four hours later animals are re-exposed to the same environment and the distance they walked is recorded. Bigger delta between days (shorter distance on the test session compared to the first session) represents intact learning.

References

Bolivar VJ, Caldarone BJ, Reilly AA, Flaherty L (2000). Habituation of activity in an open field: A survey of inbred strains and F1 hybrids. Behav Genet 30,285–293.

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Cognitive, learning and memory domain