行为实验的硬件和软件

V. Molodtsov, V. Smirnov, S. Solnushkin, V. Chikhman
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引用次数: 0

摘要

实验生理学研究的一个领域是对疼痛的研究。世界上许多实验室使用的常见疼痛刺激之一是电流。在行为实验中,对移动的实验动物施加电冲击的一种常用方法是对实验笼地板的金属条施加电压。当动物锁定具有不同电位的栅栏时,它会受到电击。在实验过程中,希望改变晶格上的电势的极性,以尽量减少动物避免电暴露的能力,位于具有相同电势的杆上。笼底金属条的极性反转顺序是这样组织的,在电流脉冲的供应周期中,没有一对铁条具有相同的极性,并且当动物关闭任何一对铁条时,每个周期至少会受到一次电击。在已知的实验技术中,施加在相邻导电棒之间的轿厢底板上的电压是通过控制电压表上的电压值的变压器手动控制的。在这种情况下,疼痛敏感性阈值的确定发生了一个错误,由于一个大的,手动可调的间距变化的电压。在许多实验中,需要提供更准确的电压电平最小值的设置和表征最小疼痛敏感阈值的电流值的注册,以及确保在不同动物的不同实验中电压电平变化步骤的准确重复性。例如,为了排除动物找到笼底等电位条以避免触电的可能性,他们对笼底的每条导电条单独施加电刺激,提供多相脉冲重复模式。为了准确有效地测定实验动物的疼痛阈值,开发并实施了MD280装置。装置中的每个电子钥匙连接到特定的寄存器位,笼底的每条通过电子钥匙连接到受控的恒压源。依次将寄存器和电压源连接到通过USB控制器连接到计算机的控制单元上。此外,该装置通过实现可调相位(通过地板导电棒上脉冲的时移设置)的脉冲馈送序列控制,消除动物避免电气冲击的可能性,并在改变参数的每一步固定和保存测量序列,从而提供正确的信息。与设备的交互使用USB 2.0 Full-Speed的计算机进行。通过USB接口与PC机的数据交换由FT245RL (FTDI)芯片上实现的USB-FIFO转换器和一个将FIFO缓冲区中的数据转换为内部命令并将数据记录到设备的FIFO缓冲区中的块执行。已经开发了RatCage程序,该程序在该设备的基础上提供了对大鼠阈值疼痛敏感性的研究。程序的图形界面允许在开始工作时选择实验参数:电刺激效果(恒定、脉冲、分相等),设置冲击参数值(电压幅值、脉冲持续时间、频率),参数如何变化(自动或手动)。施加测量信号开始后,电压幅值逐渐增大。实验者观察实验动物(大鼠)的行为,当最初的伤害性反应(惊吓)出现时,按下“记住”按钮。该程序允许您在实验数据库中保存这些值以及实验协议。由于电效应模式参数(振幅、持续时间、脉冲频率及其数量)的平滑调整,这确保了对最小疼痛敏感阈值的有效测量,当动物的主要伤害性反应发生时自动测量参数,并且避免了动物通过喂有相位的电脉冲来避免电冲击的可能性。所开发的方法和实施的工具已在ipp生理学研究所进行的疼痛研究中使用。
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Hardware and software of behavior experiments
One of the areas of experimental physiological research is the study of pain. One of the common pain stimuli used in many laboratories in the world is electric current. In behavioral experiments, a common way of organizing an electrical impact on a moving laboratory animal is to apply electrical voltage to the metal bars of the experimental cage floor. When the animal locks bars with different potential, it receives an electrical shock. It is desirable during the experiment to change the polarity of the electric potential on the bars of the lattice, in order to minimize the ability of the animal to avoid electrical exposure, located on the bars with the same electric potential. The sequence of polarity reversal on metal bars of the cage floor was organized in such a way that not a single pair of bars had the same polarity during the supply cycles of electric current pulses and the animal received an electrical shock at least once per cycle when it closed any pair of bars. Experimental techniques are known in which the voltage applied to the cage floor between adjacent conductive bars is manually controlled by means of a transformer with control of the voltage value on the voltmeter. In this case, the determination of the threshold of pain sensitivity occurs with an error due to a large, manually adjustable pitch of change in electrical voltage. In a number of experiments, it is desirable to provide a more accurate setting of the minimum values of the voltage level and the registration of the current value characterizing the minimum pain sensitivity threshold, as well as to ensure accurate repeatability of the steps of voltage level variation in different experiments with different animals. In order to exclude the possibility for animals to find equipotential bars of the cage floor to avoid electrical shock, for example, they apply electric stimulation independently to each conductive bar of the cage floor, providing a multiphase pulse repetition mode. For accurate and effective determination of the pain threshold of laboratory animals, the device MD280 was developed and implemented. Each electronic key in the device is connected to a specific register bit, and each bar of the cage floor is connected via an electronic key to a controlled constant-voltage source. In turn, the register and voltage source are connected to a control unit connected to the computer via a USB controller. In addition, the device provides correct information by eliminating the possibility of an animal avoiding electrical impact by implementing control of the pulse feed sequence with adjustable phasing (set by the time shift of the pulses on the conductive bars of the floor) and by fixing and saving the measurement sequence at each step of changing the parameters. Interaction with the device is carried out using a computer with USB 2.0 Full-Speed. Data exchange with the PC via the USB interface is performed by a USB-FIFO converter implemented on an FT245RL (FTDI) chip and a block that converts data from the FIFO buffer into internal commands and records data into the FIFO buffer from the device. The program RatCage has been developed, which provides, on the basis of the device, a study of the threshold pain sensitivity of rats. The graphical interface of the program allows at the start of work to select the parameters of the experiment: electrostimulating effects (constant, pulsed, phased, etc.), setting the values of the impact parameters (voltage amplitude, pulse duration, frequency), how parameters change (automatically or manually). After the start of the measurement signal is applied, the voltage amplitude gradually increases. The experimenter observes the behavior of the laboratory animal (rat) and when the primary nociceptive reaction (startle) appears, presses the “Remember” button. The program allows you to save these values along with the experiment protocol in the experimental database. This ensures an effective measurement of the minimum pain sensitivity threshold due to the smooth adjustment of the parameters of the electrical effect pattern (amplitude, duration, frequency of impulses, their number), automatic measurement of parameters when an animal’s primary nociceptive reaction occurs, and also the avoidance of the possibility that the animal avoids electrical impact by feeding electrical pulses with phasing. The developed approach and the implemented tools have been used in pain studies conducted at the I.P. Pavlov Institute of Physiology RAS.
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