双眼形觉剥夺对大鼠视皮层神经元电生理学和形态学特性的影响
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摘要
目的：本课题拟通过视皮层脑片膜片钳全细胞记录法，结合细胞内标记技术，观察不同发育阶段双眼形觉剥夺大鼠视皮层神经元兴奋性突触后电流(EPSCs)变化及突触传递长时程增强现象(LTP)，研究形觉剥夺对发育过程中视皮层神经元突触电生理学和形态学特性的影响，探索哺乳动物视觉发育可塑性的细胞水平机制，为临床相关视功能障碍的治疗提供新的思路。
方法：健康Wistar大鼠61只，分为正常组和双眼形觉剥夺组：正常组中3周龄（20-22天龄）13只，4周龄（27-29天龄）9只，5周龄（34-36天龄）10只；双眼形觉剥夺组3周龄（20-22天龄）12只，4周龄（27-29天龄）9只，5周龄（34-36天龄）8只。形觉剥夺组于13天时缝合双眼。制作脑片后利用盲法寻找细胞，获得膜片钳全细胞记录。通过视皮层的双极刺激电极给予0.07Hz的突触前测试刺激，记录视皮层神经元的PSCs。然后给予1Hz的低频刺激60~90个，每个脉冲伴随突触后去极化至-20mV 以诱发LTP。电生理记录结束后对所记细胞进行免疫组化染色。
结果：1、所记录的PSCs分为3种类型：无反应型、单突触反应型和多突触反应型。双眼形觉剥夺组无反应型PSCs比例（20％）显著高于正常组（3.17％）(P<0.01)；多突触反应型PSCs比例则相反，正常组（27.2％）显著高于双眼形觉剥夺组（7.94％）(P<0.01)。2、双眼形觉剥夺组与正常组间相比，输入阻抗显著增高，静息电位较为去极化，EPSCs峰值显著降低，且形态幼稚。3、形觉剥夺组LTP诱发率显著低于正常组(P<0.05)。
双眼形觉剥夺组组4周龄、5周龄的LTP增值显著高于正常组同周龄时（P<0.05）。
结论： 1、大鼠视皮层多突触反应型神经元随发育有逐渐升高的趋势，而双眼形觉剥夺后大鼠视皮层无反应型神经元较正常大鼠显著增多，提示静息突触的激活可能是视皮层神经网络回路构建的一个重要基础。
2、视皮层神经元电生理学特性和形态学特性紧密相关。形觉剥夺后大鼠视皮层神经元形态学的幼稚状态与功能上的不成熟是一致的。
3、双眼形觉剥夺后大鼠视皮层神经元LTP诱出率显著下降，提示形觉刺激是视皮层神经元突触正常发育的重要条件。
4、双眼形觉剥夺在降低视皮层神经元LTP诱发率的同时，可引起LTP幅值的明显提高。提示双眼形觉剥夺可在一定程度上降低视皮层神经元的可塑性，但可以保持神经元对适宜刺激的反应能力。
Purpose: To study electrophysiological properties of neurons in visual cortex of binocular deprivated rats during the development and explore the synaptic and cellular mechanism of experience-dependent plasticity in the visual cortex. The relationship between morphological and functional maturation of neurons in visual cortex was studied with the biocytin.
Methods: The visual cortex slices were prepared from 61 Wistar rats of both genders, which were divided into two groups: control group and binocular pattern deprivation (BD) group. In the control group, 32 rats were recorded at 3, 4 and 5 postnatal weeks. 13 rats were sacrified at 3 weeks of age (20~22days); 9 rats were sacrified at 4 weeks of age (27~29 days); 10 rats were sacrified at 5 weeks of age (34~36 days). In the BD group, 29 rats were recorded. 12 rats were sacrified at 3 weeks of age(20~22days); 9 rats were sacrified at 4 weeks of age (27~29 days) and 8 rats were sacrified at 5 weeks of age (34~36 days). Both eyelids were sutured at 13 days postnatal in BD group before the eyes open. Blind patch-clamp whole cell recording were adopted. Pre-synaptic test stimulation was given at 0.07 Hz intervals through bipolar stimulating electrodes. EPSCs of layer Ⅱ~Ⅳ were recorded by recording electrodes filled with 0.5% biocytin. Long-term potentiation (LTP) was induced by low-frequency stimulation (LFS) at 1Hz for 60~90 sec, each pulse of the LFS paired with depolarization of post-synaptic neurons to -20 mV. The neurons
were stained after recording for the purpose of morphological study.
Results: 1. There were three types of PSCs recorded: silent synaptic response, monosynaptic response, and polysynaptic response. The percentage of silent response in BD group (20%) was significantly higher than that in the control group(3.17%) (P<0.01) , While the incidence of polysynaptic EPSCs in BD group(7.94%) was significantly lower than that in the control group(27.2%) (P<0.01). 2. Compared with those of control group , neurons in BD group has a increased input resistance, peak value of EPSCs, a more depolarized resting membrane potential (P<0.05), and the neurons were morphologically less mature. 3. The incidence of LTP induced in BD group was lower while the LTP magnitude significantly higher than that in control group.
Conclusions: 1. The input resistance and the resting membrane potential are important to discriminate the mature state of the neurons.2.The incidence of polysynaptic EPSCs tends to increase gradually in control group, while there are much more silent synaptic responses in BD group. It suggests that the activation of silent synapse may relate to in formation of synaptic connections. 3. The electrophysiological and morphological properties of neurons are intimately correlated with each other. 3. The incidence of LTP induction was decreased in binocular deprivated rats suggests that the visual stimulation of patterns facilitate the maturing of the neurons in visual cortex. 5. The amplitude of LTP increased in BD group while the incidence of LTP induction was getting down compare with those of control one. It implys that the plasticity of neurons in visual cortex was decreased by BD, but the response ability to adequate stimulus still exist.
方法：健康Wistar大鼠61只，分为正常组和双眼形觉剥夺组：正常组中3周龄（20-22天龄）13只，4周龄（27-29天龄）9只，5周龄（34-36天龄）10只；双眼形觉剥夺组3周龄（20-22天龄）12只，4周龄（27-29天龄）9只，5周龄（34-36天龄）8只。形觉剥夺组于13天时缝合双眼。制作脑片后利用盲法寻找细胞，获得膜片钳全细胞记录。通过视皮层的双极刺激电极给予0.07Hz的突触前测试刺激，记录视皮层神经元的PSCs。然后给予1Hz的低频刺激60~90个，每个脉冲伴随突触后去极化至-20mV 以诱发LTP。电生理记录结束后对所记细胞进行免疫组化染色。
结果：1、所记录的PSCs分为3种类型：无反应型、单突触反应型和多突触反应型。双眼形觉剥夺组无反应型PSCs比例（20％）显著高于正常组（3.17％）(P<0.01)；多突触反应型PSCs比例则相反，正常组（27.2％）显著高于双眼形觉剥夺组（7.94％）(P<0.01)。2、双眼形觉剥夺组与正常组间相比，输入阻抗显著增高，静息电位较为去极化，EPSCs峰值显著降低，且形态幼稚。3、形觉剥夺组LTP诱发率显著低于正常组(P<0.05)。
双眼形觉剥夺组组4周龄、5周龄的LTP增值显著高于正常组同周龄时（P<0.05）。
结论： 1、大鼠视皮层多突触反应型神经元随发育有逐渐升高的趋势，而双眼形觉剥夺后大鼠视皮层无反应型神经元较正常大鼠显著增多，提示静息突触的激活可能是视皮层神经网络回路构建的一个重要基础。
2、视皮层神经元电生理学特性和形态学特性紧密相关。形觉剥夺后大鼠视皮层神经元形态学的幼稚状态与功能上的不成熟是一致的。
3、双眼形觉剥夺后大鼠视皮层神经元LTP诱出率显著下降，提示形觉刺激是视皮层神经元突触正常发育的重要条件。
4、双眼形觉剥夺在降低视皮层神经元LTP诱发率的同时，可引起LTP幅值的明显提高。提示双眼形觉剥夺可在一定程度上降低视皮层神经元的可塑性，但可以保持神经元对适宜刺激的反应能力。
Purpose: To study electrophysiological properties of neurons in visual cortex of binocular deprivated rats during the development and explore the synaptic and cellular mechanism of experience-dependent plasticity in the visual cortex. The relationship between morphological and functional maturation of neurons in visual cortex was studied with the biocytin.
Methods: The visual cortex slices were prepared from 61 Wistar rats of both genders, which were divided into two groups: control group and binocular pattern deprivation (BD) group. In the control group, 32 rats were recorded at 3, 4 and 5 postnatal weeks. 13 rats were sacrified at 3 weeks of age (20~22days); 9 rats were sacrified at 4 weeks of age (27~29 days); 10 rats were sacrified at 5 weeks of age (34~36 days). In the BD group, 29 rats were recorded. 12 rats were sacrified at 3 weeks of age(20~22days); 9 rats were sacrified at 4 weeks of age (27~29 days) and 8 rats were sacrified at 5 weeks of age (34~36 days). Both eyelids were sutured at 13 days postnatal in BD group before the eyes open. Blind patch-clamp whole cell recording were adopted. Pre-synaptic test stimulation was given at 0.07 Hz intervals through bipolar stimulating electrodes. EPSCs of layer Ⅱ~Ⅳ were recorded by recording electrodes filled with 0.5% biocytin. Long-term potentiation (LTP) was induced by low-frequency stimulation (LFS) at 1Hz for 60~90 sec, each pulse of the LFS paired with depolarization of post-synaptic neurons to -20 mV. The neurons
were stained after recording for the purpose of morphological study.
Results: 1. There were three types of PSCs recorded: silent synaptic response, monosynaptic response, and polysynaptic response. The percentage of silent response in BD group (20%) was significantly higher than that in the control group(3.17%) (P<0.01) , While the incidence of polysynaptic EPSCs in BD group(7.94%) was significantly lower than that in the control group(27.2%) (P<0.01). 2. Compared with those of control group , neurons in BD group has a increased input resistance, peak value of EPSCs, a more depolarized resting membrane potential (P<0.05), and the neurons were morphologically less mature. 3. The incidence of LTP induced in BD group was lower while the LTP magnitude significantly higher than that in control group.
Conclusions: 1. The input resistance and the resting membrane potential are important to discriminate the mature state of the neurons.2.The incidence of polysynaptic EPSCs tends to increase gradually in control group, while there are much more silent synaptic responses in BD group. It suggests that the activation of silent synapse may relate to in formation of synaptic connections. 3. The electrophysiological and morphological properties of neurons are intimately correlated with each other. 3. The incidence of LTP induction was decreased in binocular deprivated rats suggests that the visual stimulation of patterns facilitate the maturing of the neurons in visual cortex. 5. The amplitude of LTP increased in BD group while the incidence of LTP induction was getting down compare with those of control one. It implys that the plasticity of neurons in visual cortex was decreased by BD, but the response ability to adequate stimulus still exist.
引文
1. Boothe RG, Dobson MV and Teller DY. Postnatal development of vision in human and nonhuman primates. Neurosci, 8: 495-545 (1985)
2. Fox KT. he critical period for long term potentiation in primary sensory cortex. Neuron, 15:485-488(1995)
3. Sermasi, E, Tropea D. A new form of synaptic plasticity is transiently expressed in the developing rat visual cortex: Amodulatory role for visual experience and brain derived neurotrophic factor(1999)
4. Rauschecker JP. Mechanisms of visual plasticity: Hebb synapses NMDA receptors, and beyond. (1991)
5. Wiesel TN and Hubel DH. Single-cell responses in striate cortex of kittens deprived of vision in one eye. (1965)
6. Gu Q, Singer W. Effects of intracortical infusion of anticholinergic drugs on neuronal plasticity in kitten striate cortex. (1993)
7. Turlejski K, Kossut M. Decrease in the number of synapses formed by subcortical inputs to the striate cortex of binocularly deprived cats. (1985)
8. Zufferey PD, Jin F, Nakamura H. The role of pattern vision in the development of cortico-cortical connections. (1999)
9. Bliss TVP, Collingridge GL. A synaptic model of memory: long-term potentiation in the hippocampus. (1993)
10. Perkins AT and Teyler TJ. A critical period for long-term potentiation in the developing rat visual cortex. (1988)
11. Kirkwood A ,Lee HK ,Bear MF. Co-regulation of long-term potentiation and experience-dependent synaptic plasticity in visual cortex by age and experience. (1995)
MF Bear, Kleinschmidt A, Gu QA. Disruption of experience-dependent synaptic modifications in striate cortex by infusion of an NMDA receptor
12. antagonist. (1990)
13. Drager UC. Observation on monocluar deprivation on mice. J Neurophysiol. (1): 18-42 (1978)
14. Fox K. The critical period control in sensory cortex. Crru.Opin.Neurobiol, 4: 112-119 (1994)
15. Shatz CJ. Impulse activity and the patterning of connections during CNS development. Neuron, 5:745-756(1990)
16. Tepper JM, Sharp NA, Koos TZ et al. Postnatal development of the rat neostriatum: electrophysiological, light- and electron-microscopic studies. Dev Neurosci, 20: 12535-12550(1998)
17. Yingbing Liu, Peter AL, Joseph FP et al. Developmental changes in membrane properties and postsynaptic currents of granule cells in rat dentate gyrus. J Neurophysiol 76(2): 1074-1088(1996)
18. Cepeda C, Walsh JP, Buchwald NA et al. Neurophysioloical maturation of cat caudate neurons: evidence from in vitro studies. Synapse, 7: 278-290(1991)
19. Zhang L, Spigelman I and Carlen PL. Development of GABA-mediated, chloride-dependent inhibition in CA1 pyramidal neurons of immature rat hippocampal slices. J Physiol (Lond) ,444: 25-49(1991)
20. Fukuda A and Prince DA. Postnatal development of electrogenic sodium pump activity in rat hippocampal pyramidal neurons. Dev Brain Res 65: 101-114, (1992)
21. Spiegelman I, Zhang L, Carlen PL .Patch-clamp study of postnatal development of CA1 neurons in rat hippocampal slices: membrane excitability and K+ currents. J Neurophysiol. 68:55-69(1992)
22. Kirkwood A & Bear MF. J Neurosce. 14:1634-1645(1994)
23. Fregnac Y, Burkd JP, Smith DJ. Neuroyhysiol. 71: 1403-1421(1994)
Ammis CM, O'Dowd DK, Robertson RT. Activity-dependent regulation of
24. dendritic spine density on cortical pyramidal neurons in organotypic slic culture. J Neurobiol, 25: 1483(1994)
25. Cragg BG. The development of synapses in kitten visual cortex during visual deprivation. Exp Neurol, 46: 445(1975)
26. Sur C, Triller A, Korn H. Morphology of the release site of inhibitory syapses on the soma and dendrite of an identified neuron. J Comp Neurol, 351: 247-260(1994)
27. Furshpan EJ, Landis SC, Matsumoto SG. Synaptic functions in rat sympathetic neurons in microcultures. 1. Secretion of norepinephrine and acetyleholine. J Neurosci, 6: 1061-1079(1986)
28. 蔡文琴，李海标主编. 发育神经生物学. 第一版. 北京：科学出版社.116(1999)
29. Harold L. Atwood, and J. Martin Wojtowicz. Silent Synapses in Neural Plasticity: Current Evidence. Learning and memory, Vol. 6: 542-571 (1999)
30. Wang S, JM Wojtowicz and HL Atwood. Synaptic recruitment during long-term potentiation at synapses of the medial perforant pathway in the dentate gyrus of the rat brain. Synapse, 21: 78-86( 1996)
31. Malinow R. Silent synapses in three forms of central plasticity. In Central synapses: Quantal mechanisms and plasticity, pp. 226-234(1998)
32. Golshani P. and EG Jones. Synchronized paroxysmal activity in the developing thalamocortical network mediated by corticothalamic projections and "silent" synapses. J Neurosci, 19: 2865-2975 (1999)
33. Isaac JTR, RA Nicoll, and RC Malenka.. Evidence for silent synapses: Implications for the expression of LTP. Neuron, 15: 427-434(1995)
34. Grosshans DR, Clayton DA, Coultrap SJ, Browning MD. LTP leads to rapid surface expression of NMDA but not AMPA receptors in adult rat CA1. Nat Neurosci, Jan; 5(1): 27-33(2002)
35. Shao Z, Burkhalter A. Role of GABAB receptor-mediated inhibition in reciprocal interareal pathways of rat visual cortex. J Neurophysiol, 81(3): 1014-24(1999)
36. Douglas RJ，Martin KAC. A functional microcircuit for cat visual cortex. J Physiol(Lond), 440: 735-769(1991)
37. Tarczy HK, Martin LAC, Jack JJB, et al. Dynaptic interaction between smooth and spine neurons in lay 4 of cat visual cortex in vitro. J physiol, 508(2): 351-363(1998)
38. Clements JD. Transmitter timecourse in the synaptic cleft: its role in central synaptic function. Trends in Neurosci, 19: 167-171(1996)
39. Jones MV，Westbrook GZ. Desensitized states prolong GABA A channel response to brief agonist pulses. Neuron, 15: 181-191(1995)
40. Michael C. Crair, Deda C. Gillespie, Michael P. Stryker. The role of visual experience in the development of columns in cat Visual cortex. Science, vol.297: 566-570(1998)
41. Komatsu Y, Toyama K, Maeda J, et al. Long-term potentiation investigation in a slice preparation of striate cortex of young kittens. Neuroscience Letters, 26: 269-274(1981)
42. Komatsu Y，Fujii K, Maeda J, et al. Long-term potentiation of synaptic transmission in kitten visual cortex . J Neurophysiol, 59: 124-141(1988)
43. Lawrence C.Katz. What's critical for the critical period in visual cortex? Cell, vol.99: 673-676(1999)
44. Maurer D, Lewis TL, Brent HP, Levin AV. Rapid improvement in the acuity of infants after visual input. Science, Oct 1; 286(5437): 108-110(1999)
45. A.Thomas perkins Ⅳ and Timothy J.Teyler. A critical period for long-term potentiation in the developing rat visual cortex. Brain Research, 439: 222-229 (1988)
46. Harris KM and Teyler TJ. Development onset of Long-term potentiation in area CA1 of the rat hippocampus. J physiol.(London), 346: 27-48 (1984)
47. Kirkwood A.Marc, G.Rioult & Mark F.Bear. Experience-dependent modification of synaptic plasticity in visual cortex. Nature; vol. 381: 526-528(1996)
48. Buisseret P, Gary-Bobo E & Lmbert M, Nature, 272: 816-817(1978)
49. Atapour N , Esteky H , Fathollahi Y , et al . Primed-bursts induced long-term potentiation in rat visual cortex: effects of dark-rearing. Brain Res, 851: 148-153 (1999)
50. Horikawa k & Armstrong WE. A versatile means of intracellular labeling: injection of biocytin and its detection with avidin conjugates. J Neurosci Methods, 25: 1-11(1988)
51. Smithson KG, Cobbett P, Macvicar BA, et al. A reliable method for immunocytochemical identification of Lucifer Yellow injected, peptide-containing mammalian central neurons. J Nourosci Methods, 10: 59-69. (1984)
52. Tamamaki N, Watanabe K and Nojyo Y. A whole image of the hippocampal pyramidal neuron revealed by intracellular pressure-injection of horseradish peroxidase. Brain Res, 307: 336-340.(1984)
53. Zufferey PD, Jin F, Nakamura H., Tettoni L, Innocenti GM. The role of pattern vision in the development of cortico-cortical connections. Eur J Neurosci Aug; 11(8): 2669-88(1999)
54. Giese KP, Fdeorov NB, Filipkowski RK, et al. Autophosphorylation at Thr286 of the alfha calcium in LTP and learning. Science, 2779(5352): 807-813(1998)
55. Nunez A. Garcia-Austt E ,Buno W. In vivo electrophysiological analysis of Lucifer yellow-coupled hippocampal pyramids. Exp Neurol, 108: 76-82(1990)
56. Peinado A, Yuste R, Katz LC. Extensive dye coupling between rat neocortical neurons during the period of circuit formation. Neuron,10: 103-114(1993)
57. Connors BW, Bernardo LS, Prince DA. Coupling between neurons of the developing rat neocortex. J Neurosci, 3: 773-782(1983)
58. Peinado A, Yuste R, Katz LC. Extensive dye couplijng between rat neocortical neurons during the period of circuit formation. Neuron, 10: 103-114(1993)
59. Kandler K & Katz LC. Relationship between dye coupling and spontaneous activity in developing ferret visual cortex. Dev Neurosci, 20(1): 59-64(1998)
60. Tepper JM, Sharp NA, Koos TZ et al. Postnatal development of the rat neostriatum: electrophysiological, light- and electron-microscopic studies. Dev Neurosci 20: 12535-12550, 1998
61. Connors BW, Bernardo LS, Prince DA. Carbon dioxide sensitivity of dye coupling among glia and neurons of the neocortex. J Neuronsci, 4: 1324-1330(1984)
62. 丁劲，徐杏珍，李朝义 成年大鼠视皮层（17区）神经元间的染色耦合-离体脑片研究.生理学报；50（3）：241-248（1998）
2. Fox KT. he critical period for long term potentiation in primary sensory cortex. Neuron, 15:485-488(1995)
3. Sermasi, E, Tropea D. A new form of synaptic plasticity is transiently expressed in the developing rat visual cortex: Amodulatory role for visual experience and brain derived neurotrophic factor(1999)
4. Rauschecker JP. Mechanisms of visual plasticity: Hebb synapses NMDA receptors, and beyond. (1991)
5. Wiesel TN and Hubel DH. Single-cell responses in striate cortex of kittens deprived of vision in one eye. (1965)
6. Gu Q, Singer W. Effects of intracortical infusion of anticholinergic drugs on neuronal plasticity in kitten striate cortex. (1993)
7. Turlejski K, Kossut M. Decrease in the number of synapses formed by subcortical inputs to the striate cortex of binocularly deprived cats. (1985)
8. Zufferey PD, Jin F, Nakamura H. The role of pattern vision in the development of cortico-cortical connections. (1999)
9. Bliss TVP, Collingridge GL. A synaptic model of memory: long-term potentiation in the hippocampus. (1993)
10. Perkins AT and Teyler TJ. A critical period for long-term potentiation in the developing rat visual cortex. (1988)
11. Kirkwood A ,Lee HK ,Bear MF. Co-regulation of long-term potentiation and experience-dependent synaptic plasticity in visual cortex by age and experience. (1995)
MF Bear, Kleinschmidt A, Gu QA. Disruption of experience-dependent synaptic modifications in striate cortex by infusion of an NMDA receptor
12. antagonist. (1990)
13. Drager UC. Observation on monocluar deprivation on mice. J Neurophysiol. (1): 18-42 (1978)
14. Fox K. The critical period control in sensory cortex. Crru.Opin.Neurobiol, 4: 112-119 (1994)
15. Shatz CJ. Impulse activity and the patterning of connections during CNS development. Neuron, 5:745-756(1990)
16. Tepper JM, Sharp NA, Koos TZ et al. Postnatal development of the rat neostriatum: electrophysiological, light- and electron-microscopic studies. Dev Neurosci, 20: 12535-12550(1998)
17. Yingbing Liu, Peter AL, Joseph FP et al. Developmental changes in membrane properties and postsynaptic currents of granule cells in rat dentate gyrus. J Neurophysiol 76(2): 1074-1088(1996)
18. Cepeda C, Walsh JP, Buchwald NA et al. Neurophysioloical maturation of cat caudate neurons: evidence from in vitro studies. Synapse, 7: 278-290(1991)
19. Zhang L, Spigelman I and Carlen PL. Development of GABA-mediated, chloride-dependent inhibition in CA1 pyramidal neurons of immature rat hippocampal slices. J Physiol (Lond) ,444: 25-49(1991)
20. Fukuda A and Prince DA. Postnatal development of electrogenic sodium pump activity in rat hippocampal pyramidal neurons. Dev Brain Res 65: 101-114, (1992)
21. Spiegelman I, Zhang L, Carlen PL .Patch-clamp study of postnatal development of CA1 neurons in rat hippocampal slices: membrane excitability and K+ currents. J Neurophysiol. 68:55-69(1992)
22. Kirkwood A & Bear MF. J Neurosce. 14:1634-1645(1994)
23. Fregnac Y, Burkd JP, Smith DJ. Neuroyhysiol. 71: 1403-1421(1994)
Ammis CM, O'Dowd DK, Robertson RT. Activity-dependent regulation of
24. dendritic spine density on cortical pyramidal neurons in organotypic slic culture. J Neurobiol, 25: 1483(1994)
25. Cragg BG. The development of synapses in kitten visual cortex during visual deprivation. Exp Neurol, 46: 445(1975)
26. Sur C, Triller A, Korn H. Morphology of the release site of inhibitory syapses on the soma and dendrite of an identified neuron. J Comp Neurol, 351: 247-260(1994)
27. Furshpan EJ, Landis SC, Matsumoto SG. Synaptic functions in rat sympathetic neurons in microcultures. 1. Secretion of norepinephrine and acetyleholine. J Neurosci, 6: 1061-1079(1986)
28. 蔡文琴，李海标主编. 发育神经生物学. 第一版. 北京：科学出版社.116(1999)
29. Harold L. Atwood, and J. Martin Wojtowicz. Silent Synapses in Neural Plasticity: Current Evidence. Learning and memory, Vol. 6: 542-571 (1999)
30. Wang S, JM Wojtowicz and HL Atwood. Synaptic recruitment during long-term potentiation at synapses of the medial perforant pathway in the dentate gyrus of the rat brain. Synapse, 21: 78-86( 1996)
31. Malinow R. Silent synapses in three forms of central plasticity. In Central synapses: Quantal mechanisms and plasticity, pp. 226-234(1998)
32. Golshani P. and EG Jones. Synchronized paroxysmal activity in the developing thalamocortical network mediated by corticothalamic projections and "silent" synapses. J Neurosci, 19: 2865-2975 (1999)
33. Isaac JTR, RA Nicoll, and RC Malenka.. Evidence for silent synapses: Implications for the expression of LTP. Neuron, 15: 427-434(1995)
34. Grosshans DR, Clayton DA, Coultrap SJ, Browning MD. LTP leads to rapid surface expression of NMDA but not AMPA receptors in adult rat CA1. Nat Neurosci, Jan; 5(1): 27-33(2002)
35. Shao Z, Burkhalter A. Role of GABAB receptor-mediated inhibition in reciprocal interareal pathways of rat visual cortex. J Neurophysiol, 81(3): 1014-24(1999)
36. Douglas RJ，Martin KAC. A functional microcircuit for cat visual cortex. J Physiol(Lond), 440: 735-769(1991)
37. Tarczy HK, Martin LAC, Jack JJB, et al. Dynaptic interaction between smooth and spine neurons in lay 4 of cat visual cortex in vitro. J physiol, 508(2): 351-363(1998)
38. Clements JD. Transmitter timecourse in the synaptic cleft: its role in central synaptic function. Trends in Neurosci, 19: 167-171(1996)
39. Jones MV，Westbrook GZ. Desensitized states prolong GABA A channel response to brief agonist pulses. Neuron, 15: 181-191(1995)
40. Michael C. Crair, Deda C. Gillespie, Michael P. Stryker. The role of visual experience in the development of columns in cat Visual cortex. Science, vol.297: 566-570(1998)
41. Komatsu Y, Toyama K, Maeda J, et al. Long-term potentiation investigation in a slice preparation of striate cortex of young kittens. Neuroscience Letters, 26: 269-274(1981)
42. Komatsu Y，Fujii K, Maeda J, et al. Long-term potentiation of synaptic transmission in kitten visual cortex . J Neurophysiol, 59: 124-141(1988)
43. Lawrence C.Katz. What's critical for the critical period in visual cortex? Cell, vol.99: 673-676(1999)
44. Maurer D, Lewis TL, Brent HP, Levin AV. Rapid improvement in the acuity of infants after visual input. Science, Oct 1; 286(5437): 108-110(1999)
45. A.Thomas perkins Ⅳ and Timothy J.Teyler. A critical period for long-term potentiation in the developing rat visual cortex. Brain Research, 439: 222-229 (1988)
46. Harris KM and Teyler TJ. Development onset of Long-term potentiation in area CA1 of the rat hippocampus. J physiol.(London), 346: 27-48 (1984)
47. Kirkwood A.Marc, G.Rioult & Mark F.Bear. Experience-dependent modification of synaptic plasticity in visual cortex. Nature; vol. 381: 526-528(1996)
48. Buisseret P, Gary-Bobo E & Lmbert M, Nature, 272: 816-817(1978)
49. Atapour N , Esteky H , Fathollahi Y , et al . Primed-bursts induced long-term potentiation in rat visual cortex: effects of dark-rearing. Brain Res, 851: 148-153 (1999)
50. Horikawa k & Armstrong WE. A versatile means of intracellular labeling: injection of biocytin and its detection with avidin conjugates. J Neurosci Methods, 25: 1-11(1988)
51. Smithson KG, Cobbett P, Macvicar BA, et al. A reliable method for immunocytochemical identification of Lucifer Yellow injected, peptide-containing mammalian central neurons. J Nourosci Methods, 10: 59-69. (1984)
52. Tamamaki N, Watanabe K and Nojyo Y. A whole image of the hippocampal pyramidal neuron revealed by intracellular pressure-injection of horseradish peroxidase. Brain Res, 307: 336-340.(1984)
53. Zufferey PD, Jin F, Nakamura H., Tettoni L, Innocenti GM. The role of pattern vision in the development of cortico-cortical connections. Eur J Neurosci Aug; 11(8): 2669-88(1999)
54. Giese KP, Fdeorov NB, Filipkowski RK, et al. Autophosphorylation at Thr286 of the alfha calcium in LTP and learning. Science, 2779(5352): 807-813(1998)
55. Nunez A. Garcia-Austt E ,Buno W. In vivo electrophysiological analysis of Lucifer yellow-coupled hippocampal pyramids. Exp Neurol, 108: 76-82(1990)
56. Peinado A, Yuste R, Katz LC. Extensive dye coupling between rat neocortical neurons during the period of circuit formation. Neuron,10: 103-114(1993)
57. Connors BW, Bernardo LS, Prince DA. Coupling between neurons of the developing rat neocortex. J Neurosci, 3: 773-782(1983)
58. Peinado A, Yuste R, Katz LC. Extensive dye couplijng between rat neocortical neurons during the period of circuit formation. Neuron, 10: 103-114(1993)
59. Kandler K & Katz LC. Relationship between dye coupling and spontaneous activity in developing ferret visual cortex. Dev Neurosci, 20(1): 59-64(1998)
60. Tepper JM, Sharp NA, Koos TZ et al. Postnatal development of the rat neostriatum: electrophysiological, light- and electron-microscopic studies. Dev Neurosci 20: 12535-12550, 1998
61. Connors BW, Bernardo LS, Prince DA. Carbon dioxide sensitivity of dye coupling among glia and neurons of the neocortex. J Neuronsci, 4: 1324-1330(1984)
62. 丁劲，徐杏珍，李朝义 成年大鼠视皮层（17区）神经元间的染色耦合-离体脑片研究.生理学报；50（3）：241-248（1998）