大鼠单侧迷路切除术后γ-氨基丁酸A受体亚型、毒蕈碱受体亚型在前庭内侧核中的变化及毒蕈碱受体亚型在小脑绒球中的变化
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摘要
第一部分单侧迷路切除术后γ-氨基丁酸A受体亚型在大鼠前庭内侧核中的变化
目的观察左侧迷路切除术后大鼠前庭内侧核(medial vestibular nuclei, MVN)内γ-氨基丁酸A受体(gamma-aminobutyric acid A receptor, GABAA receptor)α1、β2、γ2亚型的表达变化。
方法利用免疫组织化学、原位杂交组织化学、蛋白免疫印记法、RT-PCR的方法,切除大鼠一侧迷路,观察前庭内侧核区GABAA受体α1、β2、γ2亚型表达的变化,及其在前庭代偿中可能的作用。
结果各组手术两侧比较及术后1天、3天、7天组与假手术组比较差异均无显著性意义。
结论在前庭代偿的早期,GABAA受体α1、β2、γ2亚型表达的变化极可能没有涉及前庭代偿。
第二部分单侧迷路切除术后毒蕈碱受体亚型在大鼠前庭内侧核中的变化
目的观察左侧迷路切除术后大鼠前庭内侧核(medial vestibular nuclei, MVN)内毒蕈碱受体M1、M3、M5亚型的表达变化。
方法利用免疫组织化学、RT-PCR的方法,切除大鼠一侧迷路,观察前庭内侧核区毒蕈碱受体M1、M3、M5亚型表达的变化,及其在前庭代偿中可能的作用。
结果单侧迷路切除术(UL)后可诱导双侧MVN区毒蕈碱受体M1、M3、M5亚型减少,术后1天最少,此后3天至7天处于上升趋势,7天时和假手术组比较表达无差异。各组手术两侧比较均无差异。
结论UL后可诱导MVN区毒蕈碱受体M1、M3、M5亚型减少。前庭中枢神经元的静息放电降低可能与毒蕈碱受体M1、M3、M5亚型在MVN内减少有关,但其在前庭代偿中的作用尚有待研究?
第三部分单侧迷路切除术后毒蕈碱受体亚型在大鼠小脑绒球中的变化
目的观察左侧迷路切除术后大鼠小脑绒球内毒蕈碱受体M1、M3、M5亚型的表达变化。
方法利用RT-PCR的方法,切除大鼠一侧迷路,观察小脑绒球毒蕈碱受体M1、M3、M5亚型表达的变化,及其在前庭代偿中可能的作用。
结果单侧迷路切除术(UL)后可诱导双侧绒球内毒蕈碱受体M1、M3、M5亚型减少,术后1天最少,此后3天至7天处于上升趋势,7天时和假手术组比较表达无差异。各组手术两侧比较均无差异。
结论UL后可诱导绒球内毒蕈碱受体M1、M3、M5亚型减少。前庭中枢神经元的静息放电降低可能与毒蕈碱受体M1、M3、M5亚型在绒球内减少有关,但其在前庭代偿中的作用尚有待研究?
PartⅠExpression change of GABAA receptor subunits in rat medial vestibular nucleus following unilateral labyrinthectomy
Objective To observe the expression of gamma-aminobutyric acid A receptor (GABAA receptor)α1、β2、γ2 subunits in rat medial vestibular nucleus (MVN) following left unilateral labyrinthectomy (UL). Methods The immunohistochemistry, In situ hybridization histochemistry, Western-blot and RT-PCR were used to observe the expression of GABAA receptorα1、β2、γ2 subunits post-unilateral labyrinthectomy and investigate its effect on vestibular compensation. Results No difference were observe in the ipsilateral and contralateral MVN at any group. No difference were observe in the 1 day, 3 day, 7 day than sham operation MVN following UL. Conclusion Our study suggest that expression change of GABAA receptorα1、β2、γ2 subunits in the MVN most probably were not involved in the early stage of the vestibular compensation process.
PartⅡExpression change of Muscarinic receptor subnuits in rat medial vestibular nucleus following unilateral labyrinthectomy
Objective To observe the expression of Muscarinic receptor M1、M3、M5 subunits in rat medial vestibular nucleus (MVN) following left unilateral labyrinthectomy (UL). Methods The immunohistochemistry and RT-PCR were used to observe the expression of Muscarinic receptor M1、M3、M5 subunits post-unilateral labyrinthectomy and investigate its effect on vestibular compensation. Results Muscarinic receptor M1、M3、M5 subunits were induced decrease in both side MVN after unilateral labyrinthectomy. The least expression in the 1 day MVN of following UL. The expression is rising from the 3 day to 7 day MVN of following Ul. No difference were observe in the 7 day and sham operation MVN following UL. No difference were observe in the ipsilateral and contralateral MVN at any group. Conclusions Muscarinic receptor M1、M3、M5 subunits were induced decrease in the MVN after unilateral labyrinthectomy. The fall in the resting discharge of the central vestibular neurons may be caused by the decrease of Muscarinic receptor M1、M3、M5 subunits in the MVN. But the significance of the change of Muscarinic receptor M1、M3、M5 subunits in the vestibular compensation is still unknown.
PartⅢExpression change of Muscarinic receptor subnuits in rat flocculus following unilateral labyrinthectomy
Objective To observe the expression of Muscarinic receptor M1、M3、M5 subunits in rat flocculus following left unilateral labyrinthectomy (UL). Methods The RT-PCR was used to observe the expression of Muscarinic receptor M1、M3、M5 subunits post-unilateral labyrinthectomy and investigate its effect on vestibular compensation. Results Muscarinic receptor M1、M3、M5 subunits were induced decrease in both side flocculus after unilateral labyrinthectomy. The least expression in the 1 day flocculus of following UL. The expression is rising from the 3 day to 7 day flocculus of following Ul. No difference were observe in the 7 day and sham operation flocculus following UL. No difference were observe in the ipsilateral and contralateral flocculus at any group. Conclusions Muscarinic receptor M1、M3、M5 subunits were induced decrease in the flocculus after unilateral labyrinthectomy. The fall in the resting discharge of the central vestibular neurons may be caused by the decrease of Muscarinic receptor M1、M3、M5 subunits in the flocculus. But the significance of the change of Muscarinic receptor M1、M3、M5 subunits in the vestibular compensation is still unknown.
目的观察左侧迷路切除术后大鼠前庭内侧核(medial vestibular nuclei, MVN)内γ-氨基丁酸A受体(gamma-aminobutyric acid A receptor, GABAA receptor)α1、β2、γ2亚型的表达变化。
方法利用免疫组织化学、原位杂交组织化学、蛋白免疫印记法、RT-PCR的方法,切除大鼠一侧迷路,观察前庭内侧核区GABAA受体α1、β2、γ2亚型表达的变化,及其在前庭代偿中可能的作用。
结果各组手术两侧比较及术后1天、3天、7天组与假手术组比较差异均无显著性意义。
结论在前庭代偿的早期,GABAA受体α1、β2、γ2亚型表达的变化极可能没有涉及前庭代偿。
第二部分单侧迷路切除术后毒蕈碱受体亚型在大鼠前庭内侧核中的变化
目的观察左侧迷路切除术后大鼠前庭内侧核(medial vestibular nuclei, MVN)内毒蕈碱受体M1、M3、M5亚型的表达变化。
方法利用免疫组织化学、RT-PCR的方法,切除大鼠一侧迷路,观察前庭内侧核区毒蕈碱受体M1、M3、M5亚型表达的变化,及其在前庭代偿中可能的作用。
结果单侧迷路切除术(UL)后可诱导双侧MVN区毒蕈碱受体M1、M3、M5亚型减少,术后1天最少,此后3天至7天处于上升趋势,7天时和假手术组比较表达无差异。各组手术两侧比较均无差异。
结论UL后可诱导MVN区毒蕈碱受体M1、M3、M5亚型减少。前庭中枢神经元的静息放电降低可能与毒蕈碱受体M1、M3、M5亚型在MVN内减少有关,但其在前庭代偿中的作用尚有待研究?
第三部分单侧迷路切除术后毒蕈碱受体亚型在大鼠小脑绒球中的变化
目的观察左侧迷路切除术后大鼠小脑绒球内毒蕈碱受体M1、M3、M5亚型的表达变化。
方法利用RT-PCR的方法,切除大鼠一侧迷路,观察小脑绒球毒蕈碱受体M1、M3、M5亚型表达的变化,及其在前庭代偿中可能的作用。
结果单侧迷路切除术(UL)后可诱导双侧绒球内毒蕈碱受体M1、M3、M5亚型减少,术后1天最少,此后3天至7天处于上升趋势,7天时和假手术组比较表达无差异。各组手术两侧比较均无差异。
结论UL后可诱导绒球内毒蕈碱受体M1、M3、M5亚型减少。前庭中枢神经元的静息放电降低可能与毒蕈碱受体M1、M3、M5亚型在绒球内减少有关,但其在前庭代偿中的作用尚有待研究?
PartⅠExpression change of GABAA receptor subunits in rat medial vestibular nucleus following unilateral labyrinthectomy
Objective To observe the expression of gamma-aminobutyric acid A receptor (GABAA receptor)α1、β2、γ2 subunits in rat medial vestibular nucleus (MVN) following left unilateral labyrinthectomy (UL). Methods The immunohistochemistry, In situ hybridization histochemistry, Western-blot and RT-PCR were used to observe the expression of GABAA receptorα1、β2、γ2 subunits post-unilateral labyrinthectomy and investigate its effect on vestibular compensation. Results No difference were observe in the ipsilateral and contralateral MVN at any group. No difference were observe in the 1 day, 3 day, 7 day than sham operation MVN following UL. Conclusion Our study suggest that expression change of GABAA receptorα1、β2、γ2 subunits in the MVN most probably were not involved in the early stage of the vestibular compensation process.
PartⅡExpression change of Muscarinic receptor subnuits in rat medial vestibular nucleus following unilateral labyrinthectomy
Objective To observe the expression of Muscarinic receptor M1、M3、M5 subunits in rat medial vestibular nucleus (MVN) following left unilateral labyrinthectomy (UL). Methods The immunohistochemistry and RT-PCR were used to observe the expression of Muscarinic receptor M1、M3、M5 subunits post-unilateral labyrinthectomy and investigate its effect on vestibular compensation. Results Muscarinic receptor M1、M3、M5 subunits were induced decrease in both side MVN after unilateral labyrinthectomy. The least expression in the 1 day MVN of following UL. The expression is rising from the 3 day to 7 day MVN of following Ul. No difference were observe in the 7 day and sham operation MVN following UL. No difference were observe in the ipsilateral and contralateral MVN at any group. Conclusions Muscarinic receptor M1、M3、M5 subunits were induced decrease in the MVN after unilateral labyrinthectomy. The fall in the resting discharge of the central vestibular neurons may be caused by the decrease of Muscarinic receptor M1、M3、M5 subunits in the MVN. But the significance of the change of Muscarinic receptor M1、M3、M5 subunits in the vestibular compensation is still unknown.
PartⅢExpression change of Muscarinic receptor subnuits in rat flocculus following unilateral labyrinthectomy
Objective To observe the expression of Muscarinic receptor M1、M3、M5 subunits in rat flocculus following left unilateral labyrinthectomy (UL). Methods The RT-PCR was used to observe the expression of Muscarinic receptor M1、M3、M5 subunits post-unilateral labyrinthectomy and investigate its effect on vestibular compensation. Results Muscarinic receptor M1、M3、M5 subunits were induced decrease in both side flocculus after unilateral labyrinthectomy. The least expression in the 1 day flocculus of following UL. The expression is rising from the 3 day to 7 day flocculus of following Ul. No difference were observe in the 7 day and sham operation flocculus following UL. No difference were observe in the ipsilateral and contralateral flocculus at any group. Conclusions Muscarinic receptor M1、M3、M5 subunits were induced decrease in the flocculus after unilateral labyrinthectomy. The fall in the resting discharge of the central vestibular neurons may be caused by the decrease of Muscarinic receptor M1、M3、M5 subunits in the flocculus. But the significance of the change of Muscarinic receptor M1、M3、M5 subunits in the vestibular compensation is still unknown.
引文
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1. Precht W, Dieringer N. Neuronal events paralleling functional recovery (compensation) following peripheral vestibular lesions. In: Berthoz A, Melville-Jones G, (Eds.), Adaptive mechanism in gaze control: facts and theories. Amsterdam: Elsevier, 1985:251-68.
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14. Courjon JH, Flandrin JM, Jeannerod M. et al. The role of the flocculus in vestibular compensation after hemilabyrinthectomy. Brain Research, 1982,239:251–257.
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50. Wood CD,Graybiel A. Theory of antimotion sickness drug mechanisms. Aerospace Med, 1972,43:249-252.
51. Chen K,Waller HJ,Godfrey DA. Muscarinic receptor subtypes in rat dorsal cochlear nucleus. Hear Res, 1995,89:137-145.
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3.关新民主编医学神经生物学北京:人民卫生出版社.2002
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6. de Waele C, Abitbol M, Chat M, et al.Distribution of glutamatergic receptors and GAD mRNA-containingneurons in the vestibular nuclei of normal and hemilabyrinthectomized rats. Eur J Neurosci, 1994, 6:565–576.
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10. Horii A, Kitahara T, Smith PF, et al.Effects of unilateral labyrinthectomy on GAD, GAT1, and GABA receptor gene expression in the rat vestibular nucleus.NeuroReport, 2003,18:2359–2363.
11. Adkins CE, Pillai GV, Kerby J, et al.α4β3δGABAA receptors characterized by fluorescenceresonance energy transfer-derived measurements of membranepotential. J Biol Chem, 2001.276, 38934– 38939.
12. Pirker S, Schwarze, C, Wieselthale, A., et al.GABA(A) receptors: immunocytochemical distribution of 13 subunits in the adult rat brain. Neuroscience, 2000,101:815–850.
13. Eleore L, Vassias I, Bernat I, et al. An in situ hydridization and immunofluorescence study of GABAA and GABAB receptors in the vestibular nuclei of the intact and unilaterally labyrinthectomized rat. Exp Brain Res, 2005,160:166–179.
14. Giardino L, Zanni M, Fernandez M, et al. Plasticity of GABA(a) system during ageing: focus onvestibular compensation and possible pharmacological intervention.Brain Res, 2002,929:76–86.
15. Sun, Y, Godfrey, DA, Rubin, AM. Plasticity of g-aminobutyrate in the medial vestibular nucleus of rat after inferior cerebellar peduncle transection. J Vestib Res, 2002,12:1–14.
16. Him A, Johnston AR, Yau JL, et al. Tonic activity and GABA responsiveness of medial vestibular nucleus neurons in aged rats. NeuroReport, 2001,12:3965–3968.
17. Vibert N, DeWaele C, Serafin M, et al. The vestibular system as a model of sensorimotor transformations.A combined in vivo and in vitro approach to study the cellular mechanisms of gaze and posture stabilization in mammals. Prog Neurobiol, 1997,51:243–286.
18. Vibert N, Serafin M, Vidal PP, et al. Direct and indirect effects of muscimol on medial vestibular nucleus neurones in guinea-pig brainstem slices. Exp Brain Res, 1995,104:351–356.
19. Dutia MB, Johnston AR, McQueen DS. Tonic activity of rat medial vestibular nucleus neurones in vitro and its inhibition by GABA.Exp. Brain Res, 1992,88:466–472.
20. Magnusson AK, Lindstrom S, Tham R. GABAB receptor contribute to vestibular compensation after unilateral labyrinthectomy in pigmented rats. Exp. Brain Res, 2000,134:32–41.
21. Tighilet B, Lacour M. Gamma amino butyric acid (GABA) immunoreactivity in the vestibular nuclei of normal and unilateral vestibular neurectomized cats.Eur J Neurosci, 2001,13:2255-2267.
1.关新民主编.医学神经生物学.北京:人民卫生出版社2002.
2. Koelle GB. The histochemical localization of cholinesterases in the central nervous system of the rat. J Comp Neurol, 1954,100:211-235.
3. Schwartz RD. Autoradiographic distribution of high affinity muscarinic and nicotinic cholinergic receptors labeled with [3H] acetylcholine in rat brain. Life Sci, 1986,8(23):2111-2119.
4. Burke RE,Fahn S. Choline acetyltransferase activity of the principal vestibular nuclei of rat, studies by micropunch technique. Brain Res, 1985,328(1):196-199.
5. Wamsley JK,Lewis MS,Young WS,et al. Autoradiographic localization of muscarinic cholinergic receptors in rat brain stem. J Neurosci, 1981,1:176-191.
6. Anadon R , Molist P , Rodriguez-Moldes I , et al. Distribution of cholineacetyltransferase immunoreactivity in the brain of an elasmobranch, the lesser spotted dogfish. J Comp Neurol, 2000,420(2):139-170.
7. Phelan KD,Gallagher JP. Direct muscarinic and nicotinic receptor-mediated excitation of rat medial vestibualr nucleus neurons in vitro. Synapse, 1992,10:349-358.
8. Ujihara H,Akaike A,Sasa M,et al. Muscarinic regulation of spontaneously active neurons in rat brain slices. Neurosci, Lett, 1989,106:205-210.
9. Takeshita S,Sasa M,Ishihara K,et al. Cholinergic and glutamatergic transmission in medial vestibualr neurons responding to lateral roll tilt in rats. Brain Res, 1999,840(1-2):99-105.
10. Wess J. Molecular biology of muscarinic acetylcholine receptors. Crit Rev Neurobiol, 1996, 10:69-99.
11. Wess J, Liu J, Blin N, et al. Structual basis of receptor/G protein coupling selectivity studied with muscarinic receptors as model systems. Life Sci, 1997, 60:1007-1014.
12. Onali P , Olianas MC. Bimodal regulation of cyclic AMP by muscarinic receptors:Involvement of multiple G proteins and different forms of adenylylcyclase.Life Sci, 1995,56:973-980.
13. Felder CC. Muscarinic acetylcholine receptors:Signal transduction through multiple effectors. FASEB J, 1995,9:619-625.
14. Dutia MB, Neavy P, McQueen DS, Effects of cholinergic agonists on spontaneously active rat medial vestibular neurons in vitro, J. Physiol. 1990,425:90P.
15. Sun Y, Waller HJ, Godfrey DA, et al. Spontaneous activity in rat vestibular nuclei in brain slices and effects of acetylcholine agonists and antagonists. Brain Res.2002, 934(1):58-68.
16.余其林,孔维佳.毒蕈碱受体在大鼠前庭神经核中表达的免疫组织化学与原位杂交研究.听力学及言语疾病杂志,2005,13(4):264-265.
17. Calza L, Giardino L, Zanni M, et al. Muscarinic and gamma-aminobutyric acid-ergic receptor changes during vestibular compensation. A quantitative autoradiographic study of the vestibular nuclei complex in the rat. Eur Arch Otorhinolaryngol, 1992,249:34-39.
18. Wood CD,Graybiel A. Theory of antimotion sickness drug mechanisms. Aerospace Med, 1972,43:249-252.
19. Chen K,Waller HJ,Godfrey DA. Muscarinic receptor subtypes in rat dorsal cochlear nucleus. Hear Res, 1995,89:137-145.
20. Lewis MR,Phelan KD,Shinnick GP. Primary afferent excitatory transmission recorded intracellularly in vitro from rat medial vestibular neurons. Synapse, 1989,3(2):149-153.
21. Koelle GB. Parasympathomimetic agents. In: Goodman LS,Gilman A,eds. The pharmacological basis of therapeutics. Macmillan, New York, 1975,pp:467-476.
22. Burke RE,Fahn S. Studies of somatostatin-induced barrel rotation in rats. Regul Pept, 1983,7(3):207-220.
23. Jaarsma D,Ruigrok TJ,Caffe R,et al. Cholinergic innervation and receptors in the cerebellum. Prog Brain Res, 1997,114:67-96.
24. Fukushima M,Kitahara T,Takeda N,et al. Role of cholinergic mossy fibers in medial vestibualr and prepositus hypogolssal nuclei in vestibualr compensation. Neuroscience, 2001,102(1):159-166.
25. Andre P,Pompeiano O,White SR. Role of muscarinic receptors in the cerebellar control of the vestibulospinal reflex gain: celluar mechanisms. Acta Otolaryngol Suppl, 1995,520(1):87-91.
26. Nabatame H,Sara M,Ohno Y,et al. Activation of lateral vestibular nucleus neurons by iontophoretically applied phencyclidine. J Pharmacol, 1986,42(1):117-122.
1. Precht W, Dieringer N. Neuronal events paralleling functional recovery (compensation) following peripheral vestibular lesions. In: Berthoz A, Melville-Jones G, (Eds.), Adaptive mechanism in gaze control: facts and theories. Amsterdam: Elsevier, 1985:251-68.
2. Smith P, Curthoys I. Mechanism of recovery following unilateral labyrinthectomy: a review. Brain Res Rev, 1989,14:155-80.
3. Llinas R, Walton K. Vestibular compensation: a distributed propety of the central nervous system. In: Wilson VJ, Asanuma H, eds. Integration in the nervous system. Tokyo: Igaku Shoin, 1979:145-66.
4. Johnston AR,Him A, Dutia MB. Differential regulation of GABAA and GABAB receptors during vestibular compensation. NeuroReport, 2001,12:597-600.
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