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Biology

建模生物膜与电路板，并在轴突测量电气信号：学生实验室练习

Published: January 18, 2011 doi: 10.3791/2325
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1, 1, 2, 1
1Department of Biology, University of Kentucky, 2Department of Physiology, University of Toronto

Summary

这是一个示范如何使用电气模型，是可以理解的生物膜。我们还演示程序，记录从学生为本的实验室小龙虾的腹神经索的动作电位。

Abstract

这是一个示范如何电气模型可以用来描述生物膜。这次演习还介绍了中电使用的生物物理术语。膜模型使用相同的设备在现场准备工作。调查的一个孤立的神经线的一些属性：神经的动作电位，神经元的招聘，和环境因素的反应神经线。

Discussion

我们在上线的视频演示，本文的目的是证明细胞的生物物理特性，可以部分，作为电子电路建模。此外，活的神经组织，是比较容易获得，传导速度，不应期和电生理记录技术的基本原则可能与温和的设备投资本科学生实验室。的主题和提出的基本范式，可以很容易地修改为不同课程的要求。

小龙虾和它的丰富的维护，以学生为主导的试验有吸引力的车型。甲壳动物腹神经线一般坚固，并保留最少一个小时的盐水，这是3个小时的学生实验室有足够的生理完整性。

鉴于一些小龙虾的VNC大轴突通过缝隙连接，可以进行额外的实验上所作出的贡献，以及不同的特性，比标准蛙坐骨神经准备发现可证明连接。坐骨神经是一个经典模型，为解决复合动作电位传导性能。它甚至可能是一个有趣的对比实验，让学生比较传导性能，轴突招聘，这两种制剂之间不应期。

Disclosures

没有利益冲突的声明。

Acknowledgments

从一门课程，已在使用的实验室手册，策划博士HL阿特伍德在多伦多大学动物学系，这些实验进行了修改。这次演习也被用来从制作“ 第 6 次密集IBRO基本神经科学研讨会”，并在1993年（Cooper 等人，1993年），汉城，韩国高丽大学举行的手动修改。目前的修改，需要共同使用的设备到目前在各大学的一天学生定向实验室。肯塔基大学生物系，本科课程和艺术与科学学院办公室支持。

Materials

Circuit board
  1. Electronics listed below for the breadboard experiments can be obtained at an local electronics store such as Radio Shack.
  2. Bread board, resistors, capacitors, batteries and wires that can serve as junctions for the breadboard as outlined in the experiments.
  3. A common voltmeter (Wavetek Meterman voltmeter)
  4. A/D board for on line recording to a computer. Electrical signals are recorded on line to a PowerLab/4s interface (ADInstruments, Colorado Springs, CO, USA). We use standard software from ADInstruments named Chart or Scope.

Physiology experiments
  1. Crayfish (Procambarus clarkii). Atchafalaya Biological Supply Co., Raceland, LA., USA.
  2. Standard crayfish saline: Modified from Van Harreveld's solution (1936). (in mM) 205 NaCl; 5.3 KCl; 13.5 CaCl22H2O; 2.45 MgCl26H2O; 5 HEPES and adjusted to pH 7.4. All saline chemicals were obtained from Sigma chemical company (St. Louis, MO).
  3. Dissection tools: Fine #5 tweezers, fine scissors, knife blade holder, #26002-20 insect pins (all obtained from Fine Science Tools (USA), Inc., 373-G Vintage Park Drive, Foster City, CA 94404-1139)
  4. A nerve chamber dish (ADInstruments, Colorado Springs, CO, USA) to hold the nerve. The wires that come with the nerve chamber dish can be used to stimulate the nerve.
  5. A suction electrode is used to record the signals.
  6. The manipulator or a clamp on a ring stand will also serve as a holder for the suction electrode.
  7. Faraday Cage
  8. Dissecting Microscope
  9. High Intensity Illuminator (light source)
  10. Microscope Platform
  11. AC/DC Differential Amplifier (A-M Systems Inc. Model 3000)
  12. PowerLab 26T (AD Instruments)
  13. Extracellular amplifier (AD Instruments)
  14. LabChart 7 (ADI Instruments)
  15. Dissecting tools
  16. A ring stand and clamp to serve as a holder for the recording suction electrode

DOWNLOAD MATERIALS LIST

References

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  2. Bernardini, G., Peracchia, C., Peracchia, L. L. Reversible effects of heptanol on gap junction structure and cell-to-cell electrical coupling. European Journal of Cell Biology. 34 (2), 307-312 (1984).
  3. A report on the, "SIXTH INTENSIVE IBRO WORKSHOP ON BASIC NEUROSCIENCE". Cooper, R. L., Chang, J. J., Ito, M. July 1993, Seoul, South Korea, , Society for Neuroscience. 116-116 (1985).
  4. Cragg, B. G., Thomas, P. K. The relationship between conduction velocity and the diameter and internodal length of peripheral nerve fibers. Journal of Physiology. 136, 606-614 (1957).
  5. Erlanger, J. G. asser, S, H., Bishop, G. H. The compound nature of the action current of nerves as disclosed by the cathode ray oscillograph. American Journal of Physiology. 70, 624-666 (1924).
  6. Furshpan, E. J., Potter, D. D. Transmission at the giant motor synapses of the crayfish. Journal of Physiology. 145 (2), 289-325 (1959).
  7. Johnston, M. F., Simon, S. A., Ramrn, F. Interaction of anesthetics with electrical synapses. Nature (Lond). 286, 498-500 (1980).
  8. Loewenstein, W. R. Permeability of membrane junctions. Annual NY Academy of Sciences. 137, 441-472 (1966).
  9. Meda, P., Bruzzone, R., Knodel, S., Orci, L. Blockage of cell-to-cell communication within pancreatic acini is associated with increased basal release of amylase. Journal of Cell Biology. 103 (2), 475-483 (1986).
  10. Peracchia, C. Increase in gap junction resistance with acidification in crayfish septate axons is closely related to changes in intracellular calcium but not hydrogen ion concentration. Journal of Membrane Biology. 113 (1), 75-92 (1990).
  11. Peracchia, C., Dulhunty, A. F. Low resistance junctions in crayfish: structural changes with functional uncoupling. Journal of Cell Biology. 70, 419-439 (1976).
  12. Peracchia, C., Bernardini, G., Peracchia, L. L. Is calmodulin involved in the regulation of gap junction permeability. Pfügers Arch. 399, 152-154 (1983).
  13. Peracchia, C., Lazrak, A., Peracchia, L. L. Molecular models of channel interaction and gating in gap junctions. Handbook of Membrane Channels. Molecular and Cellular Physiology. Peracchia, C. , Academic Press. San Diego. 361-377 (1994).
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  15. Spray, D. C., Harris, L. L., Bennett, M. V. L. Comparison of pH and Ca dependence of gap junctional conductance. Intracellular pH: Its Measurement, Regulation, and Utilization in Cellular Functions. Nuccitelli, R., Deamer, D. , Alan R. Liss. New York. 445-461 (1982).
  16. Spray, D. C., White, R., De Carvalho, C., Harris, A. L., Bennett, M. L. V. Gating of gap junction channels. Journal of Biophysics. 45, 219-230 (1984).
  17. Watanabe, A., Grundfest, H. Impulse propagation at the septal and commissural junctions of crayfish lateral giant axons. Journal of General Physiology. 45, 267-308 (1961).
  18. Wiersma, C. A. G., Hughes, G. M. On the functional anatomy of neuronal units in the abdominal cord of the crayfish, Procambarus clarkii. Journal of Comparative Neurology. 116, 209-228 (1961).

Tags

基本协议，第47期，无脊椎动物，小龙虾，建模，学生实验室，神经索
建模生物膜与电路板，并在轴突测量电气信号：学生实验室练习
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Cite this Article

Robinson, M. M., Martin, J. M.,… More

Robinson, M. M., Martin, J. M., Atwood, H. L., Cooper, R. L. Modeling Biological Membranes with Circuit Boards and Measuring Electrical Signals in Axons: Student Laboratory Exercises. J. Vis. Exp. (47), e2325, doi:10.3791/2325 (2011).

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