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发育中大鼠海马体中的γ-氨基丁酸A ρ受体亚基

gamma-Aminobutyric acidA rho receptor subunits in the developing rat hippocampus.

作者信息

Didelon Frédéric, Sciancalepore Marina, Savic' Natasa, Mladinic' Miranda, Bradbury Andrew, Cherubini Enrico

机构信息

Neuroscience Program and Istituto Nazionale Fisica della Materia (INFM), International School for Advanced Studies (SISSA), Trieste, Italy.

出版信息

J Neurosci Res. 2002 Mar 15;67(6):739-44. doi: 10.1002/jnr.10178.

Abstract

The RT-PCR approach was used to estimate the expression of gamma-aminobutyric acid (GABA)(A) rho receptor subunits in the hippocampus of neonatal and adult rats. All three rho subunits were detected at postnatal day (P) 2, the rho3 subunit being expressed at an extremely low level. The rho1 and rho2 products appeared to be developmentally regulated; they were found to be more pronounced in adulthood. In another set of experiments, to correlate gene expression with receptor function, GABA(A) rho subunit mRNAs were detected with single-cell RT-PCR in CA3 pyramidal cells (from P3-P4 hippocampal slices), previously characterized with electrophysiological experiments for their bicuculline-sensitive or -insensitive responses to GABA. In 6 of 19 cells (31%), pressure application of GABA evoked at -70 mV inward currents that persisted in the presence of 100 microM bicuculline (314 plus minus 129 pA). RT-PCR performed in two of these neurons revealed the presence of rho1 and rho2 subunits, the latter being present with the alpha2 subunit. A rho2 subunit was also found in 1 neuron (among 9) exhibiting a response to GABA, which was completely abolished by bicuculline. This might be due to the lack of putative accessory GABA(A) subunits that can coassemble with rho2 to make functional receptors. Similar experiments from 10 P15 CA3 pyramidal cells failed to reveal any rho1-3 transcripts. However, these neurons abundantly express alpha3 subunits. It is likely that in CA3 pyramidal cells of neonatal and adult hippocampus GABA(A) rho subunits are present but at very low levels of expression.

摘要

采用逆转录聚合酶链反应(RT-PCR)方法评估新生大鼠和成年大鼠海马中γ-氨基丁酸(GABA)(A)ρ受体亚基的表达。在出生后第2天(P2)检测到所有三种ρ亚基,其中ρ3亚基表达水平极低。ρ1和ρ2产物的表达似乎受发育调控;在成年期表达更为明显。在另一组实验中,为了将基因表达与受体功能相关联,在CA3锥体细胞(来自P3 - P4海马切片)中通过单细胞RT-PCR检测GABA(A)ρ亚基mRNA,这些细胞先前已通过电生理实验表征其对GABA的荷包牡丹碱敏感或不敏感反应。在19个细胞中的6个(31%)中,施加GABA在-70 mV时诱发内向电流,该电流在存在100 μM荷包牡丹碱时持续存在(314 ± 129 pA)。对其中两个神经元进行的RT-PCR显示存在ρ1和ρ2亚基,后者与α2亚基同时存在。在对GABA有反应的1个神经元(共9个)中也发现了ρ2亚基,该反应被荷包牡丹碱完全消除。这可能是由于缺乏可与ρ2共同组装形成功能性受体的假定辅助GABA(A)亚基。对10个P15 CA3锥体细胞进行的类似实验未能检测到任何ρ1 - 3转录本。然而,这些神经元大量表达α3亚基。很可能在新生和成年海马的CA3锥体细胞中存在GABA(A)ρ亚基,但表达水平非常低。

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