Schenk Ursula, Verderio Claudia, Benfenati Fabio, Matteoli Michela
CNR Institute of Neuroscience, Cellular and Molecular Pharmacology, Center of Excellence for Neurodegenerative Diseases, Department of Medical Pharmacology, via Vanvitelli 32, 20129 Milano, Italy.
EMBO J. 2003 Feb 3;22(3):558-68. doi: 10.1093/emboj/cdg059.
In recent years, a role for AMPA receptors as modulators of presynaptic functions has emerged. We have investigated the presence of AMPA receptor subunits and the possible dynamic control of their surface exposure at the presynaptic membrane. We demonstrate that the AMPA receptor subunits GluR1 and GluR2 are expressed and organized in functional receptors in axonal growth cones of hippocampal neurons. AMPA receptors are actively internalized upon activation and recruited to the surface upon depolarization. Pretreatment of cultures with botulinum toxin E or tetanus toxin prevents the receptor insertion into the plasma membrane, whereas treatment with alpha-latrotoxin enhances the surface exposure of GluR2, both in growth cones of cultured neurons and in brain synaptosomes. Purification of small synaptic vesicles through controlled-pore glass chromatography, revealed that both GluR2 and GluR1, but not the GluR2 interacting protein GRIP, copurify with synaptic vesicles. These data indicate that, at steady state, a major pool of AMPA receptor subunits reside in synaptic vesicle membranes and can be recruited to the presynaptic membrane as functional receptors in response to depolarization.
近年来,AMPA受体作为突触前功能调节剂的作用已逐渐显现。我们研究了AMPA受体亚基的存在情况及其在突触前膜表面暴露的可能动态调控。我们证明,AMPA受体亚基GluR1和GluR2在海马神经元轴突生长锥中表达并组装成功能性受体。AMPA受体在激活后会被主动内化,并在去极化时被招募到细胞表面。用肉毒杆菌毒素E或破伤风毒素预处理培养物可阻止受体插入质膜,而用α-银环蛇毒素处理则可增强GluR2在培养神经元生长锥和脑突触体中的表面暴露。通过可控孔径玻璃色谱法纯化小突触囊泡,发现GluR2和GluR1均与突触囊泡共纯化,但与GluR2相互作用蛋白GRIP不共纯化。这些数据表明,在稳态下,AMPA受体亚基的主要池存在于突触囊泡膜中,并且可以响应去极化作为功能性受体被招募到突触前膜。