Patent Grant
11219193
The present invention relates to a device for studying the ability of at least one mouse to orientate itself in a maze involving topographical memory and spatial memory.
The present invention relates to the field of laboratory instrumentation, and more particularly that intended for analyses of behaviour and learning in laboratory animals.
In a known manner, an early sign of the development of pathological ageing or of dementia, for example of the Alzheimer's type, is disorientation in a familiar environment. However, there is at present no behavioural procedure in animal studies that tackles characterization of the ability of such an individual to find its bearings in a familiar environment, i.e. with which the animal has previously been familiar for a considerable time.
A device is known comprising a central platform and various boxes containing food, connected to the central platform by a channel equipped with an opaque door (CN 101 884 306A).
A memory training device for animals is also known, constituted by a maze and comprising a central rotating device, connected to a motor (US 2013/0233249).
A device of the maze type for rats is also known, comprising several rows of stacked training boxes, with communication between adjacent cages, in particular by a ladder (CN 201995441 U).
There is a known document by G. Winocur et al. Neurobiology of Aging, 31 (2010), 143-150, “A study of remote spatial memory in aged rats”, in which a population of rats is introduced into a complex environment comprising a plurality of compartments making it possible to satisfy one of the rats' particular needs (playing, eating, drinking, interacting). This document does not make it possible to test the rats' orientation faculties inside the complex environment as all the compartments are visible to the rats all the time. Moreover, the various activities offered to the rats in the compartments are not recorded and no quantitative data relating to the use of said activities are measured. Finally, this test has only been described for rats and is not suitable for mice. Up to now it has been used in pharmacological studies after hippocampal lesion, but not in the context of Alzheimer's disease.
In fact, the behavioural tests used at present (tests of activity, memory, anxiety, response to stress, addiction, etc.) are strongly dependent on the animal species used, or even on the strain used of the species under consideration. Thus, very often, tests intended for rats and for mice are different, as the mouse is for example more mobile and exploratory than the rat, whereas the latter is more competent in the procedures. Measurement of complex behavioural responses therefore requires the development of new behavioural tests that are particularly suited to the species under consideration.
Various animal behaviour tests are used at present in behavioural neuropharmacology. However, none specifically addresses topographical memory in mice. The reference tests address spatial memory, such as learning in a circular swimming pool popularized by Richard Morris, or learning in a radial arm maze with eight arms that allows simultaneous analysis of errors of reference memory and of working memory. These tests have the advantage that they are used for rats and mice. However, they are essentially relevant for measuring defects in learning.
The aim of the present invention is to respond at least in large part to the above problems and moreover to provide other advantages.
Another aim of the invention is to provide directly a test usable for all the lines of transgenic mice that model Alzheimer's disease. Another aim of the invention is more effective and accurate measurement of the orientation faculties of a mouse population.
Another aim of the invention is to provide a test making it possible to characterize autism, and/or feeding behaviour, and/or social interaction and/or the sciences of movement and/or the state of depression of a mouse population.
Another aim of the invention is to offer a novel automated platform for analysing the behaviour of mice in a complex environment.
At least one of the aforementioned aims is achieved with a device for studying the topographical memory of at least one mouse, said device comprising:
It is thus possible to put a mouse population inside a special structure that will stimulate their orientation faculties, their topographical memories of said structure and more particularly of the houses in which the mice will find activities and rewards. Preferably, 6 to 8 individuals are placed in said structure.
The agora is an uncovered central space, open on at least one side to the traffic spaces of the device. The mice are placed in the agora at the start of an experiment.
The houses are located all around the agora, and at an equal distance from the latter, so that the length of the possible paths for reaching it is identical for all the houses when starting from the centre of the device. Thus, the probability that a healthy mouse, not already familiar with the device, will reach any one of the houses is identical.
A direct path is constituted by the smallest combination of streets taken for going from one point to another. A direct path is that taken by the at least one mouse for reaching a given destination, one of the houses, as quickly as possible, without making a U-turn and/or without taking a street that does not lead to the desired place.
The traffic spaces can be closed temporarily or permanently at certain positions by means of doors, in order to analyse the exploration strategy adopted by the at least one mouse.
Preferably, all the houses are located on a single level, and/or are distributed around the agora according to a star plan, each house being located at one point of said star. Advantageously, the agora is organized according to a five-pointed star plan.
More particularly, the device according to the first aspect of the invention further comprises:
The processing unit is programmed for processing the information returned by the various sensors and/or instruments and/or cameras of the device. Preferably, said returned information is processed in real time. The processing unit is programmed for recording the information collected according to a special protocol making it possible to select information of interest from all of the instrumental data. By way of non-limitative examples, the protocol allows the user to display areas of interest on the device, mouse monitoring thresholds, acquisition times, acquisition frequencies of the sensors and/or the sampling frequencies of the cameras.
Advantageously, the device can further comprise a control unit arranged for modifying the operating parameters of the sensors and/or cameras and/or light sources and/or instruments of said device. The control unit is connected to at least some of the cameras and/or light sources and/or sensors and/or instruments of said device by wire or wirelessly, and is configured for transmitting configuration parameters and/or receiving data.
The activity of the mice in the device is measured, both in the various houses and in the traffic spaces, by means of the plurality of sensors located in the device, thus making it possible to collect quantified or quantifiable data relating to the behaviour of the mice in the device. By way of non-limitative examples, the following parameters finally make it possible to characterize, in a quantifiable manner, certain aspects of the behaviour of the mice and of their faculty for orientation in the device:
They can be parameters directly measured by sensors, video monitoring or extracts of the data measured and then calculated.
For this purpose, the mice are introduced into it preferably according to a particular protocol comprising two phases:
The first phase of learning comprises several sessions of variable duration. For example, the learning phase can comprise one, four, eight or sixteen sessions of durations equal to a day, a week, two weeks or four weeks, respectively.
The second phase preferably takes place a certain time after the last learning session (for example 24 hours, 48 hours or seven days). It can comprise several sessions of tests, of variable durations and under different experimental conditions.
The experimental conditions make it possible to generate a particular craving in the mice, and in particular with respect to one of the activities or one of the stimuli that they are able to find in the device. For example, the mice can be deprived of drink or food for some hours (for example of the order of 20 h).
The mice can also be subjected to particular treatment in order to cause particular deficiencies, preferably in relation to their memory and/or their ability to find their bearings. By way of non-limitative example, the quality of the mice's topographical memory can be tested by inducing a pathological state, in the non-transgenic models of Alzheimer's disease or at an age at which transgenic animals develop the first symptoms, for example animals overexpressing mutated forms of human APP (APP: Amyloid Precursor Protein) and triggering Alzheimer's disease spontaneously.
Certain mice can also undergo the second phase without having been subjected to any stress in order to highlight certain behavioural differences.
During the second phase, the mice can be put back into the device, individually or collectively.
A device according to the invention is suitable for studying mice of any type of lines, such as C57BL/6 or Swiss, for example.
More particularly, the plurality of means of stimulation comprises at least one means for:
It is thus possible, by means of the present device, to carry out tests for characterizing autism, and/or the feeding behaviour of the mice, and/or their social interaction and/or the movement sciences and/or the state of depression of a mouse population.
Preferably, the plurality of means of stimulation can comprise a food distributor and the plurality of sensors can comprise at least one sensor arranged for measuring the volume of food eaten by at least one mouse and/or the number of usages of said food distributor and/or the time spent using it.
Moreover, the travel time for going to the house comprising the means for giving food to the at least one mouse as well as the number of errors in getting there are two parameters that are measured and calculated.
In a particular embodiment, the food distributor can comprise:
Advantageously and applicable to the various embodiments of the invention, the plurality of means of stimulation comprises at least one drink distributor and the plurality of sensors comprises at least one sensor arranged for measuring the volume of liquid drunk by the at least one mouse and/or the number of times each drink distributor is used and/or the time spent using it.
Moreover, the travel time for going to the house comprising the means for giving drink to the at least one mouse as well as the number of errors in getting there are two parameters that are measured and calculated.
In a particular embodiment, the means for giving drink to the at least one mouse can comprise:
Preferably and applicable to the various embodiments of the invention, the plurality of means of stimulation comprises a games platform comprising at least one tube arranged so that the at least one mouse can pass through it.
Advantageously, the at least one tube is made of a translucent material so that the mouse that is inside it can be seen, for example made of Plexiglas, glass or a plastic that is transparent to infrared light.
In this case the plurality of sensors can also comprise a camera for measuring the activity of the at least one mouse in the at least one tube.
Advantageously and applicable to the various embodiments of the invention, the plurality of means of stimulation comprises a running platform comprising at least one activity wheel.
In this case, the plurality of sensors can also comprise at least one sensor for measuring the number of revolutions of the wheel carried out and/or the average speed and/or the instantaneous speed and/or the usage time of each activity wheel.
Preferably and compatible with all the embodiments of the invention, the plurality of means of stimulation comprises an openwork cage of dimensions suitable for receiving another mouse and arranged to allow the at least one mouse located in the house to touch the other mouse located in said cage.
The cage is placed in one of the houses in such a way that a mouse entering the house can interact with the mouse enclosed in the cage, and preferably touch it with the end of its nose and/or with the end of at least one paw.
The cage is preferably made of a grating, the mesh of which allows establishment of this type of physical contact between mice.
In this case, the plurality of sensors can comprise a camera for measuring the interaction of the at least one mouse with the other mouse.
In particular, the following additional parameters finally make it possible to characterize, in a quantifiable manner, certain aspects of the behaviour of the mice and of their faculty for orientation in the device:
In particular and applicable to the various embodiments of the invention, the plurality of sensors comprises at least one sensor for detecting and measuring the movements of the at least one mouse inside the maze. For this purpose, said at least one sensor for detecting and measuring the movements of the at least one mouse inside the maze is arranged for detecting and tracking the shadow cast by the at least one mouse on the floor of the maze, by determining its centre of gravity for example, or its nose for more accurate tracking of its interactions with the device in a general way.
This sensor makes it possible to measure more particularly the movements of the at least one mouse in the maze and its demeanor in the maze (streets and/or houses). As explained above, these measurements, and/or those carried out for each house and for each activity, make it possible to calculate a certain number of parameters that make it possible, in particular by statistical processing of the data measured for each mouse of the plurality of mice introduced into the device, to characterize the quality of the geographical memory of such mice.
Moreover, the travel time for going to the houses comprising respectively the means for giving drink and food to the at least one mouse as well as the number of errors in getting there are two parameters that are measured and calculated.
In this case, at least a part of the floor of said device can be made of a material that is transparent to infrared radiation.
Preferably, the device can further comprise:
Preferably, the diodes, which emit in the infrared, are arranged for providing backlighting of the floor of the device according to the invention; and the infrared camera is positioned at a sufficient height to track the at least one mouse in all of the elements of the device. The device is preferably constituted by plastic that is transparent to infrared radiation.
According to another aspect of the invention, applicable with any one of the embodiments of the first aspect, a method is proposed for evaluating the ability of the at least one mouse to orientate itself in the device according to the invention [any one of the preceding claims], said method comprising:
Other advantages and characteristics of the invention will also become apparent on the one hand from the following description and on the other hand from several embodiments given by way of indication and non-limitatively with reference to the attached schematic drawings, in which:
The embodiments which will be described below are in no way limitative; it is possible in particular to imagine variants of the invention comprising only a selection of characteristics described below in isolation from the other characteristics described, if this selection of characteristics is sufficient to confer a technical advantage or to differentiate the invention with respect to the state of the prior art. This selection comprises at least one, preferably functional, characteristic without structural details, or with only a part of the structural details if this part alone is sufficient to confer a technical advantage or to differentiate the invention with respect to the state of the prior art.
In particular, all the variants and all the embodiments described can be combined with each other if there is no objection to this combination from a technical point of view.
In the figures, the elements common to several figures retain the same reference.
With reference to
The maze thus formed is preferably in the shape of a five-pointed star, in which the houses 111-115 each occupy an apex. The obstacles 141-145 are located in the middle of the arms of the star and make it possible to define in particular two traffic loops:
The outer traffic loop 161-165 is connected to the inner traffic loop 171-175 via passageways 181-185 which make it possible to avoid the obstacles 141-145.
Preferably, the outer side walls of the traffic means 161-165 forming the outer traffic loop are of a height such that the at least one mouse cannot get over it and get out of the device 100.
Advantageously, the outer side walls of the traffic means 161-165 forming the outer traffic loop are made of a translucent material so as to be able to observe the at least one mouse moving about inside and allow the animal to make use of its spatial memory on these outer traffic paths by utilizing external reference points linked to the configuration of the room and of the device and thus reinforce its cartographic and/or topographical memory of said device.
Thus, the houses 111-115 are all located at an equal distance from the agora 150 and have the same number of possible paths for reaching them from the agora 150, the direct paths all being of the same length. In other words, the houses 111-115 are arranged in such a way that the probability of reaching them from the agora 150 is the same for each of them.
The houses 111-115 are of sufficient sizes for at least one mouse to be able to enter and move about inside. In particular, they are of a sufficient height for the at least one mouse to be able to stand upright.
Advantageously, the dimensions of the sides of the houses are approximately 280 mm×160 mm, for a height of 300 mm.
Each house 111-115 has an opening 241-245 allowing the at least one mouse to enter and leave.
Each opening 241-245 of each house 111-115 opens onto a traffic space 161-165 allowing the at least one mouse to walk about in the device 100.
The device 100 according to the invention takes the form of a maze comprising the plurality of traffic spaces 161-165 and 171-175, the houses 111-115 and obstacles 141-145.
In the embodiment illustrated in
In the example illustrated in
Thus, the house 111 comprises at least one activity wheel 130, which the at least one mouse can enter and in which it can run, the wheel rotating about a fixed axis of rotation. The house 111 is called RUN in the following paragraphs.
Additionally, the house 111 can also comprise at least one sensor 131 for measuring the activity of the mouse with said activity wheel. Typically, but not limitatively, the at least one sensor 131 can be of the speed sensor type mounted on the activity wheel in order to measure the average and instantaneous rotary speed, the acceleration, a rev counter for measuring the number of revolutions made and a chronometer for measuring the duration of use of said activity wheel. A system based on the Spontaneous Activity Wheel from Bioseb can be envisaged (www.bioseb.com/bioseb/anglais/default/item_id=40.php).
The house 112 comprises a cage 122 occupying at least a part of the space of said house 112, and into which a mouse is introduced. Preferably, it is a mouse of the opposite sex to those moving about in the device 100. The cage 122 is arranged so that the mouse that is enclosed there can interact with the at least one mouse that enters the house 112. Typically, the cage 122 comprises a grating the mesh of which is sufficiently spaced for the mice to push the ends of their noses through and/or establish physical contact with each other, on either side of said cage 122. The house 112 is called INTERACT in the following paragraphs.
Additionally, the house 112 can also comprise at least one sensor 132 for measuring the activity of the mouse in said house 112, and more particularly its interaction with the mouse enclosed in the cage 122. Typically, but not limitatively, the at least one sensor 132 can be of the camera type so as to be able to study the behaviour of the mice in the house 112, and/or identify certain particular demeanours, revealing certain behavioural pathologies or deviances, for example the time and the number of direct contacts between a mouse and the animal placed in the compartment behind the grating.
The house 113 comprises at least one activity tube 123, which the at least one mouse can enter and walk in, and that it can pass through freely. Preferably, the at least one tube is located on the horizontal and/or slightly raised relative to the floor. The house 113 is called PLAY in the following paragraphs.
Additionally, the house 113 can also comprise at least one sensor 133 for measuring the activity of the mouse with the at least one activity tube 123, for example made of transparent Plexiglas. Typically, but not limitatively, the at least one sensor 133 can be of the camera type in order to study the behaviour of the at least one mouse in the tube and/or a certain particular demeanour and reveal certain behavioural pathologies or deviances, for example the time spent in the at least one tube and/or the number of times that the at least one mouse enters said tube.
The house 114 comprises a food dispenser 124 arranged so that a mouse can eat there with total autonomy. Preferably, the food dispenser 124 is of the automatic granule distributor type, comprising a main reservoir 126 for food granules, a secondary reservoir connected to the main reservoir via an automatic door that allows the secondary reservoir to be filled when it is empty or below a certain threshold. The at least one mouse can thus eat freely from the secondary reservoir. The house 114 is called EAT in the following paragraphs. The main reservoir 126 includes a feeding reservoir 127 connected to the main reservoir, and a flap 128 (motorized or not) located between the main reservoir and the feeding reservoir, the flap being arranged to open and close automatically depending on the level of food present in the feeding reservoir.
Additionally, the house 114 can also comprise at least one sensor 134 for measuring the activity of the mouse with the dispenser 124. Typically, but not limitatively, the at least one sensor 134 can be arranged for measuring the volume of food ingested by the at least one mouse at each intake, the number of times that it uses said dispenser 124 and/or its time of use. Moreover, the at least one sensor 134 also makes it possible to measure each feed intake (each mouthful), its date and time, its duration, and the quantity ingested. An automated system of the BioDAQ type from Research Diets Inc. can be used (www.researchdiets.com/biodaq/applications/food-water-intake).
Finally, the house 115 comprises at least one liquid dispenser 125 and a teat 129 located at the base of said dispenser that dispenses the liquid from the dispenser. Preferably, the house 115 comprises two dispensers 125 so as to be able to carry out measurements of hedonism, the first dispenser comprising for example a solution of sugar water and the second dispenser comprising a solution of plain water. Preferably, the dispenser 125 contains a liquid that is edible by the at least one mouse. The dispenser 125 can comprise a reservoir arranged to contain a given volume of liquid and a pipette to allow the at least one mouse to drink autonomously. The house 115 is called DRINK in the following paragraphs.
Additionally, the house 115 can also comprise at least one sensor 135 for measuring the activity of the mouse with the liquid dispenser 125. Typically, but not limitatively, the at least one sensor 135 can be arranged for measuring the volume of each portion of water taken by the at least one mouse, the number of times that it uses said dispenser 125 and/or its time of use. An automated system of the BioDAQ type from Research Diets Inc. can be used (www.researchdiets.com/biodaq/applications/food-water-intake).
As illustrated in
Advantageously, the at least one sensor 210 can be of the camera type, with a field of view at least partly covering the device 100.
In a clever way, the movement of the at least one mouse can be measured by a technique based on tracking the shadow cast by the at least one mouse on the floor 230. For this purpose, at least one lighting source 221-224 is arranged on one side of the device 100 so as to illuminate the floor of the latter uniformly.
Preferably, the at least one lighting source 221-224 is located under the floor of said device.
Advantageously, the at least one lighting source 221-224 is of the infrared source type so that the mice are not perturbed and/or influenced by the illumination of the floor 230.
In this particular embodiment, the floor 230 is transparent to the radiation emitted by the at least one lighting source 221-224, and the at least one environment sensor 210 is sensitive to infrared radiation. Thus, when the at least one mouse moves on the floor, the sensor 210 detects less light at the location of said mouse. It is therefore possible for the movement of the at least one mouse throughout the maze to be measured efficiently and at less cost.
The device also comprises a processing unit 211 that makes it possible to utilize all of the data measured by the various sensors 121-125 located in each house 111-115, as well as by the at least one environment sensor 210. The data measured are collected according to a protocol defined by the user (drawings of the areas of interest, monitoring thresholds of the animals, duration of acquisition, time intervals for collection of information, etc.). The final data are preferably presented in the form of a spreadsheet in a very versatile format to allow the user to apply their own calculation macros.
By way of non-limitative examples, the following parameters can be calculated from the data from the set of sensors 121-125, 210:
The present invention thus relates to a device for studying the ability of the at least one mouse to find its bearings in a maze, said device comprising a plurality of houses distributed around a central agora, in which means of stimulation allow the at least one mouse to carry out certain activities and/or satisfy certain primary needs. Each house further comprises at least one sensor for measuring the activity of the mouse in said house, and more particularly its use of the means of stimulation. Finally, a sensor makes it possible to detect and measure the movements of each mouse in the device. Quantitative parameters originating from these measurements thus make it possible to highlight the quality of the geographical memory of the at least one mouse.
Several examples of tests and of usage of the device according to the invention will now be described:
The mice tend to explore the various houses differently, with the highest number of entries in the house Run and the lowest scores in the house Drink. However, all the houses are properly recognized by the animals, suggesting that they explore the entire complexity of the apparatus effectively.
The results for the animals deprived of drink are presented in
As can be seen in
Familiarization in the device according to the invention therefore constitutes a rich and complex environment, which affects brain plasticity and neurogenesis of the hippocampus.
The present invention thus allows quantitative measurement of animal behaviour, in particular in the context of neuropharmacology and preclinical animal research, in particular aimed at measurement of learning processes based on the spontaneous latent exploration of a complex environment and processes of familiarization of the mice with this environment.
The present device makes it possible to measure significant differences between the behaviour of the animals with familiarization and of the animals without familiarization in an amnesia test (for water or food). The period of familiarization can vary from one day to 4 weeks or over a longer period.
The present device is able to measure the impact of cognitive deficits in particular associated with age, with pharmacological models of amnesia (treatment with scopolamine, for example), with pharmacological and genetic models of dementia (as illustrated here with the injection of amyloid peptide, but also transgenic lines of Alzheimer's disease) and other debilitating conditions (stressed or depressive animals, for example).
The present invention thus in particular makes it possible to analyse one of the important early warning signs for Alzheimer's disease: the symptom of spatial disorientation.
According to one of the embodiments, the device makes it possible to characterize autism.
According to one of the other embodiments, the device makes it possible to characterize feeding behaviour.
According to one more the other embodiments, the device makes it possible to characterize social interaction.
According to one more the other embodiments, the device makes it possible to characterize the movement sciences.
According to one more the other embodiments, the device makes it possible to characterize the state of depression of a mouse population.
Of course, the invention is not limited to the examples which have just been described and numerous adjustments can be made to these examples without exceeding the scope of the invention. In particular, the different characteristics, forms, variants and embodiments of the invention can be combined with one another according to various combinations inasmuch as they are not incompatible or mutually exclusive. In particular all the variants and embodiments described previously can be combined with each other.
| Number | Date | Country | Kind |
|---|---|---|---|
| 1557093 | Jul 2015 | FR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2016/067520 | 7/22/2016 | WO | 00 |
| Publishing Document | Publishing Date | Country | Kind |
|---|---|---|---|
| WO2017/017010 | 2/2/2017 | WO | A |
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