DOI: 10.5061/dryad.6wwpzgn8k
title: Raw data behavior in olfactory tubing maze and open field

description: Preclinical and clinical studies show that mild to moderate hypothermia is
 neuroprotective in sudden cardiac arrest, ischemic stroke, perinatal
 hypoxia/ischemia, traumatic brain injury and seizures. Induction of
 hypothermia largely involves physical cooling therapies, which induce
 several clinical complications, while some molecules have shown to be
 efficient in pharmacologically induced hypothermia (PIH). Neurotensin
 (NT), a 13 amino-acid neuropeptide that regulates body temperature,
 interacts with various receptors to mediate its peripheral and central
 effects. NT induces PIH when administered intracerebrally. However, these
 effects are not observed if NT is administered peripherally, due to its
 rapid degradation and poor passage of the blood brain barrier (BBB). We
 conjugated NT to peptides that bind the low-density lipoprotein receptor
 (LDLR) to generate “vectorized” forms of NT with enhanced BBB
 permeability. We evaluated their effects in epileptic conditions following
 peripheral administration. One of these conjugates, VH-N412, displayed
 improved stability, binding potential to both the LDLR and NTSR-1,
 rodent/human cross-reactivity and improved brain distribution. In a mouse
 model of kainate (KA)-induced status epilepticus (SE), VH-N412 elicited
 rapid hypothermia associated with anticonvulsant effects, potent
 neuroprotection and reduced hippocampal inflammation. VH-N412 also reduced
 sprouting of the dentate gyrus mossy fibers and preserved learning and
 memory skills in the treated mice. In cultured hippocampal neurons,
 VH-N412 displayed temperature-independent neuroprotective properties. To
 the best of our knowledge, this is the first report describing the
 successful treatment of SE with PIH. In all, our results show that
 vectorized NT may elicit different neuroprotection mechanisms mediated by
 hypothermia and/or by intrinsic neuroprotective properties.
description: <em>Olfactory Tubing Maze (OTM)</em>
 The hippocampus-dependent learning and memory performance of
 control mice injected with saline solution (SHAM), SE and SE + VH-N412
 mice were assessed 3 weeks after SE (12-week-old male mice). Behavioral
 assessment, based on the OTM, was carried out as we previously described
 (Roman et al., 2002; Girard et al., 2014) and we have shown that the FVB/N
 mouse strain used in the present study performed very well in this task
 (Girard et al., 2016). Briefly, mice were trained to learn odor-reward
 associations. The mice placed on water restriction were submitted to the
 presentation of odors in testing chambers (TC). Two synthetic odors
 (strawberry and jasmine), coming from 2 distinct arms and randomly
 assigned to these arms, were simultaneously presented to the mice. Each of
 these 2 odors was arbitrarily associated either with a drop of water (the
 reward) or with a non-aversive but unpleasant sound, which were delivered
 depending on the arm chosen by the mice. The OTM was composed of 4
 identical TC joined to each other forming a circular structure in which
 mice could move freely only in a clockwise direction. The procedure was
 fully automated. The movement of the mice was detected by photoelectric
 cells. Odors, water, sound delivery, and the automatic doors separating
 the different TC were controlled by a microcomputer running with LabVIEW
 software (National Instruments, Austin, TX, USA) that also automatically
 recorded the behavioral data. The learning procedure included 3
 habituation days followed by 5 days of training, each daily session being
 composed of 12 trials/odor presentations corresponding to 3 clockwise
 laps/blocks of 4 trials. Two parameters were examined: the inter-trial
 interval (ITI) and the percentage of correct responses. The ITI was
 calculated as the time between the response to an odor presentation in one
 testing chamber and the response in the next chamber. The evolution of the
 ITI reflects the fact that, after a response to an odor, the animal must
 learn to backtrack to the testing chamber and to run to the next chamber
 waiting for the opening of the entrance door for the next trial. This
 constitutes a procedural aspect of the task. The percentage of correct
 responses was the ratio of the number of correct responses to the total
 number of odor presentations per session. This percentage was used to
 evaluate the effectiveness of the association between an odor and its
 reinforcement, a process that pertains to the hippocampus-dependent
 declarative memory subcategory. <em>Open field
 test </em> As we described previously (Girard et
 al., 2016, 2014), the open field test was used to assess both exploratory
 behavior and spontaneous locomotor activity 4 weeks after SE and VH-N412
 treatment. The mice were tested using an open field square made of white
 plastic with 50 x 50 cm surface area and 45 cm-high walls. Testing was
 carried out in a dimly illuminated room (40 lux). Monitoring was done by
 an automated tracking system using a video camera mounted above the
 apparatus (Viewpoint VideoTrack version 3.0, Lyon, France). The field was
 divided virtually into 2 regions of interest: a central area (20 x 20 cm)
 and a peripheral area. The animals were placed in the center of the field
 and 2 behavioral parameters were registered during 2 consecutive 5 min
 sessions: i) the percentage of time spent in the central part versus total
 time, ii) the total traveled distance. The square was cleaned prior to the
 test and between each animal with 70% ethanol.
description: # Raw data behavior in olfactory tubing maze and open field
 [https://doi.org/10.5061/dryad.6wwpzgn8k](https://doi.org/10.5061/dryad.6wwpzgn8k) ## Description of the data and file structure Epilepsy is associated with learning and memory difficulties in patients and animal models. We thus assessed whether our hypothermia inducing molecule VH-N412-induced reduction of neurodegeneration and inflammation ameliorates mnesic capacities of our epileptic animals. Three weeks after status epilepticus, a group of mice was submitted to behavioral tests. We evaluated hippocampus-dependent learning and memory performance using the olfactory tubing maze (OTM) and the inter-trial interval (ITI). Exploratory and spontaneous locomotor activity of the same mice was assessed in the open-field paradigm where the average distances covered by mice from the different groups were evaluated. ### Files and variables #### File: Behavior\_RAW\_DATA\_OTM\_and\_OPEN\_FIELD.xlsx **Description:** Evaluate the mnesic capacities of epileptic mice treated or not with the hypothermia-inducing agent VHN-412 using 2 different tests, the olfactory tubing maze and the open field tests. ##### Variables In the olfactory tubing maze results, the inter-trial interval (ITI) is measured in seconds. In the open-field, the average distances covered by mice are measured in centimeters.