微电极矩阵研究小鼠胚胎心脏电生理活动

本实验采用一种新方法——微电极矩阵技术从整体水平研究小鼠胚胎离体整体心脏电生理活动。我们用微电极矩阵记录与60个电极相接触的心肌细胞的电活动(细胞外记录),称为场电位(field potentials,FPs),并与全细胞膜片钳记录的动作电位(action potentials,APs)(细胞内记录)进行比较,发现心房、心室处场电位形态类似于负向的细胞动作电位,场电位时程亦与动作电位时程类似。为研究兴奋的传导,我们比较了不同电极处场电位发生时间,发现在房室结构还未形成的胚胎发育第9．5天(E9．5)已经观察到明显的房室传导延迟(A-V delay)[(50．21±9．7)ms],而心室不同部位兴奋几乎是同步的。在发育晚期(E16．5),房室传导延迟为(82．21±10．50)ms。进一步研究基本的神经体液因素对心脏兴奋的调控,表明: 在E9．5,异丙肾上腺素(isoproterenol,Iso)使胚胎兴奋频率加快(34．04±7．31)％,房室传导延迟缩短(20．00±6．44)％,同时场电位时程增宽;相反,卡巴唑(carbachol,CCh)则使兴奋频率降低(42．32±5．36)％,房室传导缩短(26．00±4．81)％, 场电位时程减小。而在E16．5,Iso的作用显著增强,兴奋频率加快(101．54±10．23)％,房室传导延迟缩短(56．62±6．43)％, 而CCh则几乎使所有晚期心脏兴奋完全消失。所以,心脏的传导系统在胚胎发育早期4个腔室还未形成时已经建立,神经体液因子对心脏基本电生理活动的调控是在发育过程中逐渐成熟的。     

Search for frequency-specific effects of millimeter-wave radiation on isolated nerve function

Effects of a short-term exposure to millimeter waves (CW, 40–52 GHz, 0.24–3.0 mW/cm²) on the compound action potential (CAP) conduction were studied in an isolated frog sciatic nerve preparation. CAPs were evoked by either a low-rate or a high-rate electrical stimulation of the nerve (4 and 20 paired pulses/s, respectively). The low-rate stimulation did not alter the functional state of the nerve, and the amplitude, latency, and peak latency of CAPs could stay virtually stable for hours. Microwave irradiation for 10–60 min at 0.24–1.5 mW/cm², either at various constant frequencies or with a stepwise frequency change (0.1 or 0.01 GHz/min), did not cause any detectable changes in CAP conduction or nerve refractoriness. The effect observed under irradiation at a higher field intensity of 2–3 mW/cm² was a subtle and transient reduction of CAP latency and peak latency along with a rise of the test CAP amplitude. These changes could be evoked by any tested frequency of the radiation; they reversed shortly after cessation of exposure and were both qualitatively and quantitatively similar to the effect of conventional heating of 0.3–0.4°C. The high-rate electrical stimulation caused gradual and reversible decrease of the amplitude of conditioning and test CAPs and increased their latencies and peak latencies. These changes were essentially the same with and without irradiation (2.0–2.7 or 0.24–0.28 mW/cm²), except for attenuation of the decrease of the test CAP amplitude. This effect was observed at both field intensities, but was statistically significant only for certain frequencies of the radiation. Within the studied limits, this effect appeared to be dependent on the frequency rather than on the intensity of the radiation, but this observation requires additional experimental confirmation. Bioelectromagnetics 18:324–334, 1997. © 1997 Wiley-Liss, Inc.     

蛙离体单根有髓鞘神经纤维静息电位与动作电位细胞内记录方法

本文介绍一种记录蛙离体单根有髓鞘神经纤维的静息和动作电位的细胞内记录方法,包括神经干标本的制作和固定、微电极和刺激电极的制备、简易防震和静息与动作电位的记录.该方法记录的静息电位和动作电位幅值,在30min 内可保持相对稳定。     

低温保存许旺细胞对周围神经再生的作用

目的：比较原代培养许旺细胞（Schwann cells,SCs)和冷冻保存的SCs移植对损伤后坐骨神经再生的作用。方法：原代培养和液氮保存的SCs分别移植到桥接缺损坐骨神经的硅胶管内。在移植后不同时间（第6和8周末）,硅胶管远端神经干内注射HRP,逆行追踪背根神经节和脊髓前角的标记神经元数量;测量再生神经纤维的复合动作电位传导速度;电镜观察再生神经纤维的髓鞘形成。结果：原代培养和冷冻保存SCs在移植后不同时间其背根神经节和脊髓前角神经元HRP标记细胞数量、再生神经纤维的复合动作电位传导速度基本一致,再生神经纤维髓鞘的形成未见明显差别。结论：冷冻保存的SCs仍具有促进损伤后周围神经再生的能力。     

Effect of static magnetic fields on bioelectric properties of the Br and N1 neurons of snail Helix pomatia

The effects of 2.7 mT and 10 mT static magnetic fields were investigated on two identified neurons with different bioelectric properties of the snail Helix pomatia. Membrane resting potential, amplitude, spiking frequency, and duration of action potential were measured. The two neurons of H. pomatia, parabolic burster Br and silent N₁, showed different responses to a static magnetic field. The magnetic field of 2.7 mT intensity caused changes in the amplitude and duration of action potential of the Br neuron, whereas the 10 mT magnetic field changed the resting potential, amplitude spike, firing frequency, and duration of action potential of the Br neuron. Bioelectric parameters measured on the N₁ neuron did not change significantly in these magnetic fields.     

催产素对豚鼠听觉机能作用的电生理研究

本实验利用听觉电生理学方法,研究了催产素(Oxytocin)对豚鼠内耳听觉机能的作用。给豚鼠肌内注射催产素后,由短声引起的耳蜗微音器电位和听神经复合动作电位幅值增加,听神经复合动作电位和听皮层诱发电位的阈值降低。说明催产素具有提高豚鼠内耳听觉机能的作用。     

牛蛙神经冲动产生和传导的低温效应

在5℃和15℃温度条件下,用牛蛙(Rana catesbeiana)离体坐骨神经标本测定0、24、48、96、120、144、168、192、216 h 9个时段的动作电位波幅和传导速度.结果表明:两个温度下离体坐骨神经的动作电位幅度在0 h和24 h差异均不显著,0 h时相对高温(15℃)下动作电位传导速度大于相对低温(5℃),24 h时两个温度下动作电位的传导速度差异不显著,相对高温下48 h时坐骨神经的兴奋性为零.相对低温条件下,坐骨神经兴奋性能维持7 d时间.     

Influence of weakened constant magnetic field on nerve cell excitability

Attenuation of the constant magnetic field by a factor of 200–250 (to ～0.2 μT) raised the excitation threshold and decreased the amplitude of the action potentials in the isolated frog sciatic nerve, and altered the conformation of carotenoids in the plasma membrane (sciatic axolemma) as well as in intracellular vesicles (cytosomes of snail ganglia). The variation in nerve excitability was supposedly caused by changes in the ordering of membrane lipids.     

50-Hz magnetic field exposure system for small animals

The design, construction, and results of evaluation of an animal-exposure system for the study of biological effects of extremely low frequency (ELF) magnetic fields are described. The system uses a square coil arrangement based on a modification of the Helmholtz coil. Due to the cubic configuration of this exposure system, horizontal and vertical magnetic fields as high as 0.3 mT can be generated. Circularly polarized magnetic fields can also be generated by changing the current and phase difference between two sets of coils. Tests were made for uniformity of the magnetic field, stray fields, sham-exposure ratio of stray field, changes of temperature and humidity, light intensity and distribution inside the animal-housing space, and noise due to air-conditioning equipment. Variation of the magnetic field was less than 2% inside the animal housing. The stray-field level inside the sham-exposure system is less than 2% of experimental exposure levels. The system can be used for simultaneous exposure of 48 rats (2 to a cage) or 96 mice (4 to a cage). © 1993 Wiley-Liss. Inc.     

青风藤水煎液对蟾蜍坐骨神经干电生理的影响

目的：探索青风藤对蟾蜍离体坐骨神经干电生理特性的影响。方法：40只蟾蜍随机分成4组：任氏液对照组和青风藤低、中、高剂量组（0.2、0.10、0.05 g/ml）（n=10）,通过RM6240C多道生理信号采集处理系统记录其在不同浓度青风藤水煎分别浸泡15 min和30 min时,动作电位的传导速度、幅度和动作电位阈强度。结果：与对照组相比,高剂量组的神经干动作电位的传导速度显著减慢（P<0.01）,中、高剂量组的幅度显著降低（P<0.01）,高剂量组的动作电位阈强度显著增大（P<0.01）。结论：动作电位传导速度、幅度与青风藤剂量呈负相关,动作电位阈强度与青风藤剂量呈正相关。青风藤水煎液降低坐骨神经的兴奋性,阻滞动作电位的传导,可能发挥抑制坐骨神经痛的作用。     

Effects of static magnetic field on specific adenosine-5′-triphosphatase activities and bioelectrical and biomechanical properties in the rat diaphragm muscle

In this study, we aimed to clarify the effects of chronically applied static magnetic field (200 Gauss) on specific ATPase activities and bioelectrical and biomechanical responses in the isolated rat diaphragm muscle. The mean activities of Na⁺-K⁺ ATPase and Ca²⁺ ATPase determined from the diaphragm homogenates were significantly higher in the magnetic field exposed group (n = 20), but that of Mg²⁺ ATPase was nonsignificantly lower compared to the control group (n = 13). Resting membrane potential, amplitude of muscle action potential, and overshoot values (mean ± SE) in the control group were found to be ?76.5 ± 0.6, 100 ± 0.8, and 23.5 ± 0.6 mV, respectively; these values were determined to be ?72.8 ± 0.4, 90.3 ± 0.5, and 17.2 ± 0.4 mV in the magnetic field-exposed group, respectively. The latency was determined to increase in the experimental group, and all the above-mentioned bioelectrical differences between the groups were significant statistically. Force of muscle twitch was found to decrease significantly in the magnetic field-exposed group, and this finding was attributed to the augmenting effect of magnetic field on Ca²⁺ ATPase activity. These results suggest that magnetic field exposure changes specific ATPase activities and, thence, bioelectrical and biomechanical properties in the rat diaphragm muscle. © 1995 Wiley-Liss, Inc.     

Variable spatial magnetic field influences peripheral nerves regeneration in rats

Generator of spatial magnetic field is one of most recent achievements among the magnetostimulators. This apparatus allows to obtain the rotating magnetic field. This new method may be more effective than other widely used techniques of magnetostimulation and magnetotherapy. We investigated the influence of alternating, spatial magnetic field on the regeneration of the crushed rat sciatic nerves. Functional and morphological evaluations were used. After crush injury of the right sciatic nerve, Wistar C rats (n?=?80) were randomly divided into four groups (control and three experimental). The experimental groups (A, B, C) were exposed (20?min/day, 5?d/week, 4 weeks) to alternating spatial magnetic field of three different intensities. Sciatic Functional Index (SFI) and tensometric assessments were performed every week after nerve crush. Forty-eight hours before the sacrificing of animals, DiI (1,1’-di-octadecyl-3,3,3’,3’-tetramethyloindocarbocyanine perchlorate) was applied 5?mm distally to the crush site. Collected nerves and dorsal root ganglia (DRG) were subjected to histological and immunohistochemical staining. The survival rate of DRG neurons was estimated. Regrowth and myelination of the nerves was examined. The results of SFI and tensometric assessment showed improvement in all experimental groups as compared to control, with best outcome observed in group C, exposed to the strongest magnetic field. In addition, DRG survival rate and nerve regeneration intensity were significantly higher in the C group. Above results indicate that strong spatial alternating magnetic field exerts positive effect on peripheral nerve regeneration and its application could be taken under consideration in the therapy of injured peripheral nerves.     

草甘膦胁迫对中华大蟾蜍（Bufo gargarizans）神经冲动产生和传导的影响

应用电生理方法研究了除草剂草甘膦对中华大蟾蜍（Bufo gargarizans Cantor）坐骨神经干冲动产生和传导的影响。用不同浓度的草甘膦溶液对中华大蟾蜍进行胁迫处理,草甘膦有效成分经由皮肤进入蟾蜍体内而作用于神经系统,利用生物信号采集处理系统测定草甘膦胁迫下中华大蟾蜍离体坐骨神经干的应激反应时间、动作电位幅度和冲动传导速度,结果表明：随着草甘膦溶液浓度的升高,中华大蟾蜍坐骨神经干接受刺激后产生冲动所需的时间逐渐延长,动作电位峰值降低,神经冲动传导速度亦逐渐减慢。草甘膦施用后,中华大蟾蜍7d内的平均应激反应时间与草甘膦浓度呈正相关,而动作电位幅度及传导速度均与草甘膦浓度呈负相关。草甘膦溶液浓度达到推荐农田使用浓度1.64~2.87ml/L时,各处理组蟾蜍的应激反应时间、动作电位幅度和冲动传导速度均与对照组差异极显著（P〈0.01）。同时,随着试验处理时间的延长,中华大蟾蜍神经干对刺激的反应变得更为迟钝,神经冲动的传导速度也进一步减慢。回归分析可知,中华大蟾蜍坐骨神经干的应激反应时间与草甘膦施用后天数呈正相关,而神经传导速度与药后天数呈负相关。由此可以说明,草甘膦胁迫条件下,中华大蟾蜍神经细胞对刺激反应的灵敏性降低,动作电位的产生及传导受到一定程度的抑制和阻碍。     

Measurement and analysis of static magnetic fields that block action potentials in cultured neurons

To characterize the properties of static magnetic fields on firing of action potentials (AP) by sensory neurons in cell culture, we developed a mathematical formalism based on the expression for the magnetic field of a single circular current loop. The calculated fields fit closely the field measurements taken with a Hall effect gaussmeter. The biological effect induced by different arrays of permanent magnets depended principally on the spatial variation of the fields, quantified by the value of the gradient of the field magnitude. Magnetic arrays of different sizes (macroarray: four center-charged neodymium magnets of ?14 mm diameter; microarray: four micromagnets of the same material but of ?0.4 mm diameter) allowed comparison of fields with similar gradients but different intensities at the cell position. These two arrays had a common gradient value of ?1 mT/mm and blocked >70% of AP. Alternatively, cells placed in a field strength of ?0.2 mT and a gradient of ?0.02 mT/mm produced by the macroarray resulted in no significant reduction of firing; a microarray field of the same strength but with a higher gradient of ?1.5 mT/mm caused ?80% AP blockade. The experimental threshold gradient and the calculated threshold field intensity for blockade of action potentials by these arrays were estimated to be ?0.02 mT/mm and ?0.02 mT, respectively. In conclusion, these findings suggest that spatial variation of the magnetic field is the principal cause of AP blockade in dorsal root ganglia in vitro. © 1995 Wiley-Liss, Inc.     

低镁和缺镁对离体豚鼠右心室肌单细胞动作电位的效应

取豚鼠右心室肌,在改良 Krebs 溶液灌注下,用微电极记录动作电位(AP)12只豚鼠72次心肌单细胞 AP 有关参数的平均值为:静息电位(RP)-76±9mV;动作电位振幅(APA)107±7mV;动作电位时程(APD)_(_30mv)为254±123ms;APD_(100)为312±133ms。当灌注液中镁离子浓度减低到0.6mol/L 时,72次 AP 的 APD_(_30mv)和APD_(100)分別为对照值的80.7%和83%;在无镁溶液中,改变更为显著,分别为对照值的70.9%和76.7%;RP 和 APA 则变化均不大。实验提示:低镁可使 APD 缩短,从而可能影响体表心电图 T 波的第3位相;此外,APD 的缩短意味着不应期相对缩短,这或许是低镁症时出现室性早搏的因素。