CN109173056B - A rat spinal nerve signal detection and evaluation system and method - Google Patents
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Abstract
本发明公开了一种大鼠脊髓神经信号检测与评估系统及方法，包括刺激电极、记录电极、脉冲刺激器和神经信号采集系统，刺激电极连接脉冲刺激器，记录电极连接神经信号采集系统，本发明利用电子学检测方法对大鼠的脑初级运动皮层进行功能电刺激，记录脊椎T9‑T12节段对应的脊髓神经信号和相应位点坐标，从相关系数和延迟时间两个指标检测大鼠脑运动皮层支配下肢运动的神经在脊髓中的传导通路，结果表明有效位点记录到的信号之间有着较高的相关系数，且存在一定的延时。本发明能够为微电子神经桥接实验的探测电极植入提供实用数据，从电子信息学角度对大鼠脊髓传导功能进行检测和评估，在动物实验水平证明了微电子神经桥接实验的可行性。
The invention discloses a rat spinal cord nerve signal detection and evaluation system and method, comprising a stimulation electrode, a recording electrode, a pulse stimulator and a nerve signal acquisition system. The stimulation electrode is connected to the pulse stimulator, and the recording electrode is connected to the nerve signal acquisition system. The invention uses an electronic detection method to perform functional electrical stimulation on the primary motor cortex of the rat's brain, records the spinal cord nerve signals corresponding to the T9-T12 segment of the spine and the coordinates of the corresponding site, and detects the rat brain from the two indicators of the correlation coefficient and the delay time. The motor cortex controls the conduction pathway of the nerves in the spinal cord for lower extremity movement. The results show that there is a high correlation coefficient between the signals recorded at the effective site, and there is a certain delay. The invention can provide practical data for the implantation of probe electrodes in the microelectronic nerve bridging experiment, detect and evaluate the conduction function of the rat spinal cord from the perspective of electronic informatics, and prove the feasibility of the microelectronic nerve bridging experiment at the level of animal experiments.
Description
Technical Field
The invention relates to the technical field of nerve signal detection and evaluation, in particular to a rat spinal cord nerve signal detection and evaluation system and method.
Background
The spinal cord is a part of the central nervous system of human and vertebrate animals, and inside the vertebral canal, the upper end is connected with medulla oblongata, and the two sides send out paired nerves which are distributed to four limbs, body wall and viscera. The inside of the spinal cord is provided with an H-shaped (butterfly-shaped) gray matter area which is mainly composed of nerve cells; the periphery of the gray matter region is a white matter region, which is mainly composed of myelinated nerve fibers. The spinal cord is the central pivot for many simple reflexes. A number of paired nerves (called spinal nerves) are sent from both sides of the spinal cord and distributed throughout the body to the skin, muscles and internal organs. The spinal cord is the pathway between the peripheral nerves and the brain. And is also a low-level central pivot for many simple reflex activities. Spinal cord injury is usually complicated by spinal trauma. In severe cases, spinal cord injury can cause paralysis of the lower extremities, incontinence of urine and feces, etc. The spinal cord is located in the spinal canal, is cylindrical, slightly deviated from the front to the back, and is enveloped by a capsule, which is consistent with the curvature of the spinal column. The upper end of the spinal cord is connected with the medulla oblongata at the level of the occipital foramen, and the lower end of the spinal cord is level with the lower edge of the first lumbar vertebra and is about 40-45 cm long. The end of the spinal cord becomes thinner, called the spinal column. Down the spinal column is an elongated terminal filament, which is already a thin plexus of denervated tissue, wrapped in dura mater at the level of the second sacral vertebra, ending down the back of the coccyx. The whole length of the spinal cord is different in thickness, two dilatations are provided, and the cervical dilatation is called from the fourth segment of the cervical vertebra to the first segment of the thoracic vertebra; the expansion of the waist is called from the ninth to twelfth nodes of the thoracic vertebrae. The surface of the spinal cord has two median longitudinal grooves, front and back, which are divided into two symmetrical halves. The anterior median fissure is deeper in the anterior aspect and the posterior median sulcus is shallower in the posterior aspect. Two pairs of lateral sulci are also present, namely the anterolateral sulci and the posterolateral sulci. The anterior root emerges from the anterolateral sulcus and consists of motor nerve fibers; the posterior root enters the spinal cord through the posterolateral sulcus and consists of the central processes of the ganglion sensory neurons. Each posterior root has an enlarged spinal ganglion before it meets the anterior root. The lumbar, sacral, and anterior and posterior roots of the caudal region travel a long distance down the spinal canal around the terminal filaments before passing through the corresponding intervertebral foramina, which collectively form the cauda equina. In adults (men), generally, the spinal cord is absent below the first lumbar vertebra and only the cauda equina is present; a transverse section of the spinal cord showing gray matter in the central part and white matter in the peripheral part; the cervical spinal cord, gray matter and white matter are well developed.
With the development of economic levels of countries in the world, the incidence of spinal cord injury tends to increase year by year. Spinal cord injury is the most serious complication of spinal cord injury, often leading to severe dysfunction of the limb below the injured segment. The spinal cord injury not only brings serious physical and psychological damage to patients, but also causes huge economic burden to the whole society. Due to the socio-economic loss caused by spinal cord injury, prevention, treatment and rehabilitation of spinal cord injury have become a major topic in the medical community today. Spinal cord injury is a trauma to the central nervous system and is one of the major causes of disability in humans. Approximately 20-40 people per million have spinal cord injuries that can cause motor and sensory impairment, loss of labor and living abilities, and significant mental injury. The scholars try to achieve the purpose of curing the spinal cord injury by stem cell transplantation, bionic scaffold materials, nerve growth inhibition factor elimination and other methods, and although certain effect is achieved, a plurality of adverse factors and treatment technology are difficult. Therefore, how to effectively treat spinal cord injuries remains a significant challenge. The early treatment of spinal injuries comprises first aid, emergency treatment, early special treatment and the like. The correctness of early treatment measures directly influences the life safety of patients and the recovery of spinal and spinal functions. Early assessment of patients with various wounds should be performed from the wound site. Patients with reduced consciousness or coma often fail to complain of pain. The possibility of spinal injury is suspected for any patient with craniocerebral injury, severe facial or scalp laceration and multiple injuries, and further injury to nervous tissues is reduced through orderly rescue and transportation.
Rat, a collective term for various rodents in the murine family, often refers to various unrelated species in the family. The artificial feeding is started in the later stage of the 18 th century, habit of the artificial feeding is that the artificial feeding is carried out day and night, the artificial feeding is loved to live alone, the gallbladder is small and frightened, incisors are long, gnawing and biting are favored, disease resistance is strong, sensitivity is strong, genetics is consistent, irritability and attack are easy to bite hands when catching, especially, female mice in the lactation period are more fierce, and the hands which are stretched into a mouse cage when workers feed are often bitten actively. Rats respond to experimental conditions more closely, are known as precise biological research tools, are widely used for research on aspects such as endocrine, medicine, ethology, geriatrics, tumors, infectious diseases, cardiovascular diseases, traditional Chinese medicine and the like, have multiple varieties and strains, and can be selected for different experiments.
With the further crossing and fusion of electronic technology and neuroscience, the university of southeast university biological electronics expert of Wangzhanggong function professor, the university of biomedical engineering expert of Ludao and the university of southeast university neuroscience expert of Judao firstly propose to reconstruct the spinal function by adopting a microelectronic bridging mode. The method aims to realize the bridging, signal regeneration and function recovery of nerve channels at two ends of a spinal cord injury plane by means of an implanted microelectronic chip, namely, the microelectronic system is used for establishing physical bridging for a central nervous system interrupted after injury so as to realize the bidirectional conduction of nerve signals, thereby realizing the reconstruction of the central nervous function. Based on the scheme, a conduction function detection experiment of rat brain primary motor cortical nerves in the spinal cord based on functional electrical stimulation is carried out and evaluated.
Disclosure of Invention
The present invention is directed to a system and a method for detecting and evaluating rat spinal cord neural signals, so as to solve the problems mentioned in the background art.
In order to achieve the purpose, the invention provides the following technical scheme: a rat spinal cord neural signal detection and evaluation system comprises stimulation electrodes, recording electrodes, a pulse stimulator and a neural signal acquisition system, wherein the stimulation electrodes are respectively connected with the pulse stimulator, and the recording electrodes are connected with the neural signal acquisition system.
Preferably, the stimulating electrode adopts a tungsten wire single electrode of American MicroProbes, and has the electrode model of WE30030.5A3, the shaft diameter of 0.081mm and the tip diameter of 2-3 μm.
Preferably, the recording electrode is a tungsten wire single electrode manufactured by MicroProbes, USA, with model number of WE30031.0A3, axial diameter of 0.081mm, and tip diameter of 2-3 μm.
Preferably, the pulse stimulator adopts a Master-9 pulse stimulator, the frequency of the pulse stimulator is 1Hz, the wave width of the pulse stimulator is 1ms, and the magnitude of the stimulation current is adjusted through an isolator.
Preferably, the neural signal acquisition system has a sampling frequency of 10kHz, analog band-pass filtering of 0.1-1kHz, a digital notch of 50Hz and an amplification factor of 50.
Preferably, the using method comprises the following steps:
A. the full-automatic brain stereotaxic apparatus is used for performing functional electrical stimulation on a primary motor cortex of a rat brain, and simultaneously detecting the cross section of a spinal T9 segment;
B. the point with the largest signal amplitude is selected as the effective point,
C. after the effective sites are determined, keeping the three-dimensional coordinates of the recording electrodes in the T9 spinal segment spinal cord of the vertebra unchanged, moving the other recording electrode to the tail end, and carrying out signal detection on the T10, T11 and T12 spinal segments spinal cords of the vertebra according to the same method;
D. simultaneously recording evoked potentials of two sites in the spinal cord, detecting and evaluating correlation and time delay between signals by a cross-correlation function analysis method, ensuring that the signals come from the same signal source, and connecting the recording sites to form a conduction path of the rat brain primary motor cortex nerve signals in the spinal cord.
Preferably, in the step A, the implantation site of the stimulation electrode is 2.00mm after the coronary suture, the sagittal suture is 2.00mm to the right and is 2.00mm deep, and the implantation site is a rat brain primary motor cortex.
Preferably, in the step C, the implantation site of the recording electrode uses the junction of the posterior median sulcus of the spinal cord and the spinal segment as an origin, and uses the transverse diameter direction as an X axis, the dorsal-abdominal direction as a Y axis, and the head-tail direction as a Z axis.
Compared with the prior art, the invention has the beneficial effects that: the invention utilizes an electronic detection method to carry out functional electrical stimulation on the brain primary motor cortex of a rat, records spinal cord nerve signals (evoked potentials) and corresponding site coordinates corresponding to spinal T9-T12 segments, and detects the conduction path of the nerves of the rat brain motor cortex governing the movement of lower limbs in the spinal cord from two indexes of correlation coefficients and delay time. The experimental result shows that the signals recorded by the effective sites have higher correlation coefficients and have certain time delay. The higher correlation coefficient reflects that the signals recorded by the two sites are most likely to originate from the same signal source, namely the cortex site stimulated in the experiment. The delay time reflects the conduction velocity of the signal when the distance between the two recording sites is fixed, and the conduction velocity is within a reasonable range to prove that the detected signal is a neural signal and not a stimulation artifact. The spinal nerve conduction velocity measured in the experiment approximately coincides with 10-50m/s reported by Mccomas. Because different anesthetic drugs and anesthetic depths can cause different influences on nerve conduction velocity and functions, the conduction velocity measured in the experiment is only used as a judgment basis for detecting whether the conduction velocity is a stimulation artifact or not, and provides a reference for measuring the nerve signal conduction velocity of the rat under the anesthesia with the same dose. The experiment solves the problem of detecting electrode implantation in the microelectronic nerve bridging scheme, and provides practical data for reconstructing motor function lost due to spinal cord injury by microelectronic technology.
Drawings
FIG. 1 is a schematic diagram of the system of the present invention;
FIG. 2 is a T9 and T10 segmental spinal cord evoked potential map of rat spine of the present invention;
FIG. 3 is a graph of the correlation analysis of spinal evoked potentials of the T9 and T10 segments of rat vertebrae;
FIG. 4 is a T9 and T11 segmental spinal cord evoked potential map of rat vertebrae of the present invention;
FIG. 5 is a graph of the correlation analysis of spinal evoked potentials of the T9 and T11 segments of rat vertebrae;
FIG. 6 is a T9 and T12 segmental spinal cord evoked potential map of rat vertebrae of the present invention;
FIG. 7 is a graph of the correlation analysis of spinal evoked potentials of the T9 and T12 segments of rat vertebrae.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1, the present invention provides a technical solution: a rat spinal cord neural signal detection and evaluation system comprises a stimulation electrode 1, a recording electrode 2, a pulse stimulator 3 and a neural signal acquisition system 4, wherein the stimulation electrode 1 is connected with the pulse stimulator 3, and the recording electrode 2 is connected with the neural signal acquisition system 4. The stimulating electrode 1 adopts a tungsten wire single electrode of American MicroProbes, the model of the electrode is WE30030.5A3, the axial diameter is 0.081mm, and the diameter of the tip is 2-3 mu m; the recording electrode 2 adopts a tungsten wire single electrode produced by American MicroProbes, the model of the electrode is WE30031.0A3, the axial diameter is 0.081mm, and the diameter of the tip is 2-3 μm; the pulse stimulator 3 adopts a Master-9 pulse stimulator, the frequency of the pulse stimulator is 1Hz, the wave width is 1ms, and the magnitude of the stimulation current is adjusted by an isolator; the neural signal acquisition system 4 has the sampling frequency of 10kHz, the analog band-pass filtering of 0.1-1kHz, the digital notch of 50Hz and the amplification factor of 50.
The using method of the invention comprises the following steps:
A. the full-automatic brain stereotaxic apparatus is used for performing functional electrical stimulation on a primary motor cortex of a rat brain, and simultaneously detecting the cross section of a spinal T9 segment;
B. the point with the largest signal amplitude is selected as the effective point,
C. after the effective sites are determined, keeping the three-dimensional coordinates of the recording electrodes in the T9 spinal segment spinal cord of the vertebra unchanged, moving the other recording electrode to the tail end, and carrying out signal detection on the T10, T11 and T12 spinal segments spinal cords of the vertebra according to the same method;
D. simultaneously recording evoked potentials of two sites in the spinal cord, detecting and evaluating correlation and time delay between signals by a cross-correlation function analysis method, ensuring that the signals come from the same signal source, and connecting the recording sites to form a conduction path of the rat brain primary motor cortex nerve signals in the spinal cord.
In the step A, the implantation site of the stimulating electrode is 2.00mm behind a coronary seam, the sagittal seam is 2.00mm to the right side, and the depth is 2.00mm, and the implantation site is a rat brain primary motor cortex; and C, taking the junction of the posterior median sulcus of the spinal cord and the spinal segment as an origin point, taking the transverse radial direction as an X axis, taking the dorsal-abdominal direction as a Y axis, and taking the head-tail direction as a Z axis.
Spinal conduction function tests were performed on spinal cords corresponding to spinal T9 and T10 segments of 6 SD rats. The electrode implantation site coordinates were recorded and normalized, and the results are given in the following table:
the T9 and T10 spinal cord evoked potentials of primary rat vertebrae are shown in figure 2.
The cross-correlation function analysis of the spinal cord evoked potentials of the T9 and T10 segments of the rat spine shows that the correlation coefficient is 0.951 as shown in figure 3, which indicates that the signal correlation degree of the two channels is highly correlated. Recording site spacing of 4.68mmAnd the nerve signal is delayed by 0.1ms, and the corresponding spinal nerve conduction velocity is calculated to be 46.8m/s, which indicates that the signals measured by the recording electrode come from the same signal source, namely the primary motor cortex of the brain and are not stimulation artifacts.
Spinal conduction function tests were performed on spinal cords corresponding to spinal T9 and T11 segments of 6 SD rats. The electrode implantation site coordinates were recorded and normalized, and the results are given in the following table:
the T9 and T11 spinal cord evoked potentials of primary rat vertebrae are shown in FIG. 4.
The cross-correlation function analysis of the T9 and T11 spinal cord evoked potentials of rat vertebrae is performed, and the result is shown in fig. 5, wherein the correlation coefficient is 0.7276, which indicates that the signal correlation degree of the two channels is significant correlation. The distance between the recording points is 8.96mm, the nerve signal is delayed for 0.2ms, and the corresponding spinal nerve conduction velocity is 44.8m/s by calculation, which indicates that the signals measured by the recording electrodes come from the same signal source, namely the primary motor cortex of the brain and are not stimulation artifacts.
Spinal conduction function tests were performed on spinal cords corresponding to spinal T9 and T12 segments of 6 SD rats. The electrode implantation site coordinates were recorded and normalized, and the results are given in the following table:
the T9 and T12 spinal cord evoked potentials of primary rat vertebrae are shown in FIG. 6.
The cross-correlation function analysis of the T9 and T11 spinal cord evoked potentials of rat vertebrae is performed, and the result is shown in fig. 7, wherein the correlation coefficient is 0.7593, which indicates that the signal correlation degree of the two channels is significant correlation. The interval of recording sites is 15.26mm, the nerve signal is delayed for 0.5ms, and the corresponding spinal nerve conduction velocity is calculated to be 30.52m/s, which indicates that the signals measured by the recording electrodes come from the same signal source, namely the primary motor cortex of the brain and are not stimulation artifacts.
The invention utilizes an electronic detection method to carry out functional electrical stimulation on the brain primary motor cortex of a rat, records spinal cord nerve signals (evoked potentials) and corresponding site coordinates corresponding to spinal T9-T12 segments, and detects the conduction path of the nerves of the rat brain motor cortex governing the movement of lower limbs in the spinal cord from two indexes of correlation coefficients and delay time. The experimental result shows that the signals recorded by the effective sites have higher correlation coefficients and have certain time delay. The higher correlation coefficient reflects that the signals recorded by the two sites are most likely to originate from the same signal source, namely the cortex site stimulated in the experiment. The delay time reflects the conduction velocity of the signal when the distance between the two recording sites is fixed, and the conduction velocity is within a reasonable range to prove that the detected signal is a neural signal and not a stimulation artifact. The spinal nerve conduction velocity measured in the experiment approximately coincides with 10-50m/s reported by Mccomas. Because different anesthetic drugs and anesthetic depths can cause different influences on nerve conduction velocity and functions, the conduction velocity measured in the experiment is only used as a judgment basis for detecting whether the conduction velocity is a stimulation artifact or not, and provides a reference for measuring the nerve signal conduction velocity of the rat under the anesthesia with the same dose. The experiment solves the problem of detecting electrode implantation in the microelectronic nerve bridging scheme, and provides practical data for reconstructing motor function lost due to spinal cord injury by microelectronic technology.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
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