多巴胺受体在大鼠嗅球的表达及其在帕金森病大鼠嗅觉障碍中的作用
|
摘要
目的观察多巴胺受体在大鼠嗅球(OB)的表达与分布,探讨左旋多巴(L-DOPA)治疗对帕金森病(PD)大鼠嗅觉的影响。方法采用免疫印迹、免疫荧光等方法观察多巴胺受体在大鼠OB中的表达; 6-羟多巴胺(6-OHDA)双侧注射建立PD大鼠模型,检测L-DOPA治疗对PD大鼠嗅觉功能及谷氨酸脱羧酶(GAD)和脑源性神经营养因子(BNDF)表达的影响。结果嗅球内D1和D2两种多巴胺受体亚型表达含量高。D1和D2在颗粒细胞层(GCL)内GAD阳性的γ-氨基丁酸(GABA)能神经元上大量表达,被酪氨酸羟化酶(TH)阳性神经纤维终末包绕。PD大鼠OB内GCL层TH蛋白表达明显下降(0. 05±0. 01 vs 0. 01±0. 00,P<0. 001)。L-DOPA治疗后,PD大鼠找寻食物小球时间显著降低[(624. 4±113. 4) s vs(312. 4±79. 35) s,P<0. 05],OB内BDNF表达显著升高(0. 02±0. 01 vs0. 07±0. 01,P<0. 01)。结论 D1和D2在GCL层GABA能神经元大量表达。L-DOPA治疗可缓解PD大鼠嗅觉障碍,可能与激活OB内GABA能神经元上的D1和D2复合体,进而改善BDNF表达有关。
Objective To observe the localization and expression of dopamine receptors in olfactory bulb(OB) of rats and explore the effect of L-levodopa(L-DOPA) treatment on hyposmia in Parkinson's disease(PD) rat model.Methods Western blotting,immunohistochemistry and immunofluorescence were used to observe the expression and localization of dopamine receptors in the OB. PD rat model was established by bilateral 6-hydroxy dropamine(6-OHDA)injection to detect the effect of L-DOPA treatment on the hyposmia and the expression of glutamic decarboxylase(GAD)and brain derived neurotrophic factor(BNDF) of PD rats. Results D1 and D2 receptors were the major subtypes in the OB.D1 and D2 receptors were expressed by GAD positive γ-aminobutyric acid(GABA) ergic neurons in the granule cell layer(GCL) which surrounded by tyrosine hydroxylase(TH) positive nerve fibers. The expression of TH in the GCL layer of PD rats decreased significantly(0. 05±0. 01 vs 0. 01± 0. 00,P< 0. 001). After L-DOPA treatment,the time of finding food balls in PD rats was significantly reduced [(624. 4±113. 4) s vs(312. 4± 79. 35) s,P< 0. 05]and the expression of BDNF in the OB was increased(0. 02 ± 0. 01 vs 0. 07 ± 0. 01,P < 0. 01). Conclusion D1 and D2 are expressed in the GABAergic neurons in the GCL layer of OB. L-DOPA treatment alleviates the hyposmia of PD rats,which may be related to the D1-D2 receptor heteromeractivation and its downstream BDNF expression of GABAergic neurons.
Objective To observe the localization and expression of dopamine receptors in olfactory bulb(OB) of rats and explore the effect of L-levodopa(L-DOPA) treatment on hyposmia in Parkinson's disease(PD) rat model.Methods Western blotting,immunohistochemistry and immunofluorescence were used to observe the expression and localization of dopamine receptors in the OB. PD rat model was established by bilateral 6-hydroxy dropamine(6-OHDA)injection to detect the effect of L-DOPA treatment on the hyposmia and the expression of glutamic decarboxylase(GAD)and brain derived neurotrophic factor(BNDF) of PD rats. Results D1 and D2 receptors were the major subtypes in the OB.D1 and D2 receptors were expressed by GAD positive γ-aminobutyric acid(GABA) ergic neurons in the granule cell layer(GCL) which surrounded by tyrosine hydroxylase(TH) positive nerve fibers. The expression of TH in the GCL layer of PD rats decreased significantly(0. 05±0. 01 vs 0. 01± 0. 00,P< 0. 001). After L-DOPA treatment,the time of finding food balls in PD rats was significantly reduced [(624. 4±113. 4) s vs(312. 4± 79. 35) s,P< 0. 05]and the expression of BDNF in the OB was increased(0. 02 ± 0. 01 vs 0. 07 ± 0. 01,P < 0. 01). Conclusion D1 and D2 are expressed in the GABAergic neurons in the GCL layer of OB. L-DOPA treatment alleviates the hyposmia of PD rats,which may be related to the D1-D2 receptor heteromeractivation and its downstream BDNF expression of GABAergic neurons.
引文
[1]Linster C, Cleland TA. Neuromodulation of olfactory transformations[J]. Curr Opin Neurobiol,2016,40:170-177.
[2]Dang LL,Tan J. Research progress on the regulating effect of dopamine in olfactory bulbneural circuits[J]. Chinese Journal of Clinical Neurosciences,2017,25(1):77-80.(in Chinese)党琳琳,谭洁.多巴胺对嗅球神经回路调节作用的研究进展[J].中国临床神经科学,2017,25(1):77-80.
[3]Cheng W,Ren Zh H,Guan YQ,et al. Impact on the dopaminergic neurons of olfactory bulb incynomolgus monkeys damaged by MPTPP[J]. Acta Anatomica Sinica,2014,45(3):333-337.(in Chinese)程炜,任振华,关云谦,等. 1-甲基-4-苯基-1,2,3,6-四氢吡啶对食蟹猴嗅球多巴胺能神经元的影响[J].解剖学报,2014,45(3):333-337.
[4]Hglinger GU,Alvarez-Fischer D,Arias-Carrión O,et al. A new dopaminergic nigro-olfactory projection[J]. Acta Neuropathol,2015,130(3):333-348.
[5]Zhang YM,Lian H,Song HY,et al. Changes of ionized calcium binding adapter molecule 1 and pro-opiome-lanocortin expression in arcuate nucleus of hypothalamus induced by substantia nigra lesion with 6-hydroxydopamine[J]. Chinese Journal of Pathophysiology,2017,33(6):975-980.(in Chinese)张毅敏,连辉,宋海岩,等. 6-羟多巴损伤黑质导致弓状核离子钙接头蛋白1和阿片-促黑素皮质素原表达的变化[J].中国病理生理杂志,2017,33(6):975-980.
[6]Chiu WH,Depboylu C,Hermanns G,et al. Long-term treatment with L-DOPA or pramipexole affects adult neurogenesis and corresponding non-motor behavior in a mouse model of Parkinson’s disease[J]. Neuropharmacology,2015,95:367-376.
[7]Liu Y, Sun JD, Song LK, et al. Environment-contact administration of rotenone:a new rodent model of Parkinson’s disease[J]. Behav Brain Res,2015,294:149-161.
[8]CoronasⅤ,Srivastava LK,Liang JJ,et al. Identification and localization of dopamine receptor subtypes in rat olfactory mucosa and bulb:a combined in situ hybridization and ligand binding radioautographic approach[J]. J Chem Neuroanat,1997,12(4):243-257.
[9]Vijayanathan Y,Lim FT,Lim SM,et al. 6-OHDA-lesioned adult zebrafish as a useful Parkinson’s disease model for dopaminergic neuroregeneration[J].Neurotox Res,2017,32(3):496-508.
[10]Todd KL, Baker KL, Eastman MB, et al. EphA4 regulates neuroblast and astrocyte organization in a neurogenic niche[J]. J Neurosci,2017,37(12):3331-3341.
[11]O’Keeffe GC,Barker RA,Caldwell MA,et al. Dopaminergic modulation of neurogenesis in the subventricular zone of the adult brain[J]. Cell Cycle,2009,8(18):2888-2894.
[12]Hglinger GU,Rizk P,Muriel MP,et al. Dopamine depletion impairs precursor cell proliferation in parkinson disease[J]. Nat Neurosci,2004,7(7):726-735.
[13]Sui Y,Horne MK,Stanic'DO’Keeffe,et al. Reduced proliferation in the adult mouse subventricular zone increases survival of olfactory bulb interneurons[J]. PLoS One,2012,7(2):e31549.
[14]Hasbi A,Fan T,Alijaniaram M,et al. Calcium signaling cascade links dopamine D1-D2 receptor heteromer to striatal BDNF production and neuronal growth[J]. Proc Natl Acad Sci USA,2009,106(50):21377-21382.
[15]Perreault ML,Fan T,Alijaniaram M,et al. Dopamine D1-D2receptor heteromer in dual phenotype GABA/glutamatecoexpressing striatal medium spiny neurons:regulation of BDNF,GAD67 and VGLUT1/2[J].PLoS One,2012,7(3):e33348.
[16]Mc Dole B,Isgor C,Pare CPerreault,et al. BDNF over-expression increases olfactory bulb granule cell dendritic spine density in vivo[J]. Neuroscience,2015,304:146-160.
[2]Dang LL,Tan J. Research progress on the regulating effect of dopamine in olfactory bulbneural circuits[J]. Chinese Journal of Clinical Neurosciences,2017,25(1):77-80.(in Chinese)党琳琳,谭洁.多巴胺对嗅球神经回路调节作用的研究进展[J].中国临床神经科学,2017,25(1):77-80.
[3]Cheng W,Ren Zh H,Guan YQ,et al. Impact on the dopaminergic neurons of olfactory bulb incynomolgus monkeys damaged by MPTPP[J]. Acta Anatomica Sinica,2014,45(3):333-337.(in Chinese)程炜,任振华,关云谦,等. 1-甲基-4-苯基-1,2,3,6-四氢吡啶对食蟹猴嗅球多巴胺能神经元的影响[J].解剖学报,2014,45(3):333-337.
[4]Hglinger GU,Alvarez-Fischer D,Arias-Carrión O,et al. A new dopaminergic nigro-olfactory projection[J]. Acta Neuropathol,2015,130(3):333-348.
[5]Zhang YM,Lian H,Song HY,et al. Changes of ionized calcium binding adapter molecule 1 and pro-opiome-lanocortin expression in arcuate nucleus of hypothalamus induced by substantia nigra lesion with 6-hydroxydopamine[J]. Chinese Journal of Pathophysiology,2017,33(6):975-980.(in Chinese)张毅敏,连辉,宋海岩,等. 6-羟多巴损伤黑质导致弓状核离子钙接头蛋白1和阿片-促黑素皮质素原表达的变化[J].中国病理生理杂志,2017,33(6):975-980.
[6]Chiu WH,Depboylu C,Hermanns G,et al. Long-term treatment with L-DOPA or pramipexole affects adult neurogenesis and corresponding non-motor behavior in a mouse model of Parkinson’s disease[J]. Neuropharmacology,2015,95:367-376.
[7]Liu Y, Sun JD, Song LK, et al. Environment-contact administration of rotenone:a new rodent model of Parkinson’s disease[J]. Behav Brain Res,2015,294:149-161.
[8]CoronasⅤ,Srivastava LK,Liang JJ,et al. Identification and localization of dopamine receptor subtypes in rat olfactory mucosa and bulb:a combined in situ hybridization and ligand binding radioautographic approach[J]. J Chem Neuroanat,1997,12(4):243-257.
[9]Vijayanathan Y,Lim FT,Lim SM,et al. 6-OHDA-lesioned adult zebrafish as a useful Parkinson’s disease model for dopaminergic neuroregeneration[J].Neurotox Res,2017,32(3):496-508.
[10]Todd KL, Baker KL, Eastman MB, et al. EphA4 regulates neuroblast and astrocyte organization in a neurogenic niche[J]. J Neurosci,2017,37(12):3331-3341.
[11]O’Keeffe GC,Barker RA,Caldwell MA,et al. Dopaminergic modulation of neurogenesis in the subventricular zone of the adult brain[J]. Cell Cycle,2009,8(18):2888-2894.
[12]Hglinger GU,Rizk P,Muriel MP,et al. Dopamine depletion impairs precursor cell proliferation in parkinson disease[J]. Nat Neurosci,2004,7(7):726-735.
[13]Sui Y,Horne MK,Stanic'DO’Keeffe,et al. Reduced proliferation in the adult mouse subventricular zone increases survival of olfactory bulb interneurons[J]. PLoS One,2012,7(2):e31549.
[14]Hasbi A,Fan T,Alijaniaram M,et al. Calcium signaling cascade links dopamine D1-D2 receptor heteromer to striatal BDNF production and neuronal growth[J]. Proc Natl Acad Sci USA,2009,106(50):21377-21382.
[15]Perreault ML,Fan T,Alijaniaram M,et al. Dopamine D1-D2receptor heteromer in dual phenotype GABA/glutamatecoexpressing striatal medium spiny neurons:regulation of BDNF,GAD67 and VGLUT1/2[J].PLoS One,2012,7(3):e33348.
[16]Mc Dole B,Isgor C,Pare CPerreault,et al. BDNF over-expression increases olfactory bulb granule cell dendritic spine density in vivo[J]. Neuroscience,2015,304:146-160.