The expression of dominant-negative subunits selectively suppresses neuronal AMPA and kainate receptors.
The expression of dominant-negative subunits selectively suppresses neuronal AMPA and kainate receptors.
复制标题
显性失活亚基的表达选择性抑制神经元 AMPA 和红藻氨酸受体。
DOI：
10.1016/s0306-4522(02)00534-1
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发表时间：
2002
期刊：
影响因子：
3.3
通讯作者：
Howe,JamesR
中科院分区：
文献类型：
--
作者：
Robert,Antoine;Hyde,Rhonda;Hughes,ThomasE;Howe,JamesR
Glutamate-gated ion channels are widely expressed in neurons where they serve a host of cellular functions. An appealing, but yet unexplored, way to delineate the functions of particular glutamate receptor subtypes is to direct the expression of dominant-negative and gain-of-function mutant subunits. We tested the ability of two dominant-negative subunits, an α-amino-3-hydroxy-5-methyl-isoxazolproprionic acid receptor subunit and a kainate receptor subunit, to silence recombinant and neuronal glutamate receptors. Co-expression studies in non-neuronal cells indicated that the inclusion of a single mutant subunit was sufficient to silence the receptor. When expressed in cerebellar granule cells, the dominant-negative subunits silenced native channels in a subtype-specific fashion. Immunocytochemical staining of control and transfected neurons, as well as studies with a gain-of-function glutamate receptor-1 mutant, indicated that the mutant subunits were expressed at levels roughly equal to the total abundance of related native subunits, and both dominant-negatives suppressed native channel expression 60–65% when tested 24 h post-transfection. If co-assembly of the mutant subunits with related native subunits is combinatorial, this level of suppression gives receptor half-lives of approximately 20 h.
