Giraldez L, Díaz-Hernández M, Gómez-Villafuertes R, Pintor J, Castro E, Miras-Portugal M T
Departamento de Bioquímica y Biología Molecular IV, Facultad de Veterinaria, Universidad Complutense de Madrid, Madrid, Spain.
J Neurosci Res. 2001 Apr 15;64(2):174-82. doi: 10.1002/jnr.1063.
Synaptosomal preparations from rat midbrain exhibit specific responses to both ATP and Ap(5)A, which stimulate a Ca(2+) increase in the presynaptic terminals via specific ionotropic receptors, termed P2X, and diadenosine polyphosphate receptors. Aminergic terminals from rat brain basal ganglia were characterized by immunocolocalization of synaptophysin and the vesicular monoamine transporter VMAT2 and represent 29% of the total. These aminergic terminals respond to ATP and/or Ap(5)A with an increase in the intrasynaptosomal calcium concentration as measured by a microfluorimetric technique. This technique, which allows single synaptic terminals to be studied, showed that roughly 8.2% +/- 1.6% of the aminergic terminals respond to ATP, 16.9% +/- 1.3% respond to Ap(5)A, 32.6% +/- 0.8% to both, and 42.3% +/- 1.5% of them have no response. Immunological studies performed with antibodies against ionotropic ATP receptor subunits showed positive labelling with anti-P2X(3) antibodies in 39% of the terminals. However, colocalization studies of VMAT and P2X(3) receptor subunit indicate that only 25% of the aminergic terminals also contain this receptor subtype. These results demonstrate that the aminergic terminals from the rat brain basal ganglia are to a large extent under the modulation of presynaptic nucleotide and dinucleotide receptors.
大鼠中脑的突触体制剂对ATP和Ap(5)A均表现出特异性反应,它们通过特定的离子otropic受体(称为P2X)和二腺苷多磷酸受体刺激突触前终末内[Ca(2+)]i升高。大鼠脑基底神经节的胺能终末通过突触素和囊泡单胺转运体VMAT2的免疫共定位进行表征,占总数的29%。这些胺能终末对ATP和/或Ap(5)A的反应是突触体内钙浓度升高,这是通过微荧光技术测量的。该技术允许研究单个突触终末,结果显示约8.2%±1.6%的胺能终末对ATP有反应,16.9%±1.3%对Ap(5)A有反应,32.6%±0.8%对两者都有反应,42.3%±1.5%无反应。用抗离子otropic ATP受体亚基的抗体进行的免疫学研究显示,39%的终末用抗P2X(3)抗体呈阳性标记。然而,VMAT和P2X(3)受体亚基的共定位研究表明,只有25%的胺能终末也含有这种受体亚型。这些结果表明,大鼠脑基底神经节的胺能终末在很大程度上受突触前核苷酸和二核苷酸受体的调节。