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离子型谷氨酸受体亚基在金鱼视网膜中的表达与定位——原位杂交和免疫细胞化学研究

Expression and localization of ionotropic glutamate receptor subunits in the goldfish retina--an in situ hybridization and immunocytochemical study.

作者信息

Vandenbranden C A, Kamphuis W, Nunes Cardozo B, Kamermans M

机构信息

The Netherlands Ophthalmic Research Institute, Meibergdreef 47, 1105 BA Amsterdam, The Netherlands.

出版信息

J Neurocytol. 2000 Oct;29(10):729-42. doi: 10.1023/a:1010938420762.

Abstract

The expression and distribution of AMPA, kainate and NMDA glutamate receptor subunits was studied in the goldfish retina. For the immunocytochemical localization of the AMPA receptor antisera against GluR2, GluR2/3 and GluR4 were used, and for in situ hybridization rat specific probes for GluR1 and GluR2 and goldfish specific probes for GluR3 and GluR4 were used. The localization of the low affinity kainate receptor and NMDA receptor was studied using antisera against GluR5-7 and NR1. All AMPA receptor subtypes were demonstrated to be present in the goldfish retina both by immunocytochemistry and in situ hybridization. In situ hybridization revealed expression of all AMPA receptors subunit at the inner border of the INL. Only GluR3 was also strongly expressed in the outer border of the INL. Some of the ganglion cells displayed a strong signal for GluR1, GluR3 and GluR4. GluR1-immunoreactivity was present in subsets of bipolar, amacrine, and ganglion cells. GluR2 and GluR2/3-immunoreactivity was mainly localized in the outer plexiform layer. GluR2 and GluR2/3-immunoreactivity are associated with the photoreceptor synaptic terminals. GluR4-immunoreactivity is present on Müller cells in the inner retina and on dendrites of bipolar cells in the OPL, whereas GluR5-7-immunoreactivity was prominently present on horizontal cell axon terminals. Finally, NR1-immunoreactivity was confined to amacrine cells, the inner plexiform layer and ganglion cells. This study shows that there is a strong heterogeneity of glutamate receptor subunit expression in the various layers of the retina. Of the AMPA receptor subunits GluR3 seems to be expressed the most widely in all layers with strong glutamatergic synaptic interactions whereas all the other subunits seem to have a more restricted expressed pattern.

摘要

在金鱼视网膜中研究了AMPA、海人酸和NMDA谷氨酸受体亚基的表达与分布。针对AMPA受体进行免疫细胞化学定位时,使用了抗GluR2、GluR2/3和GluR4的抗血清,而原位杂交则使用了大鼠特异性的GluR1和GluR2探针以及金鱼特异性的GluR3和GluR4探针。使用抗GluR5 - 7和NR1的抗血清研究了低亲和力海人酸受体和NMDA受体的定位。通过免疫细胞化学和原位杂交均证实金鱼视网膜中存在所有AMPA受体亚型。原位杂交显示所有AMPA受体亚基在INL的内边界表达。只有GluR3在INL的外边界也有强烈表达。一些神经节细胞对GluR1、GluR3和GluR4显示出强信号。GluR1免疫反应性存在于双极细胞、无长突细胞和神经节细胞的亚群中。GluR2和GluR2/3免疫反应性主要定位于外网状层。GluR2和GluR2/3免疫反应性与光感受器突触终末相关。GluR4免疫反应性存在于内视网膜的Müller细胞以及OPL中双极细胞的树突上,而GluR5 - 7免疫反应性显著存在于水平细胞轴突终末上。最后,NR1免疫反应性局限于无长突细胞、内网状层和神经节细胞。这项研究表明,视网膜各层中谷氨酸受体亚基的表达存在很强的异质性。在AMPA受体亚基中,GluR3似乎在所有层中表达最广泛,具有强烈的谷氨酸能突触相互作用而所有其他亚基似乎具有更局限的表达模式。

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