High and low GABA sensitivity α4β2δ GABA_A receptors are expressed in Xenopus laevis oocytes with divergent stoichiometries

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• 作者：Leonny Y. Hartiadi ; Philip K. Ahring ; Mary Chebib¹ ; Nathan L. Absalom ; ¹ ; ^{nathan.absalom@sydney.edu.au" class="auth_mail" title="E-mail the corresponding author}
• 关键词：GABAA receptor ; γ-amino butyric acid receptor type A ; DS ; Delta Selective ; THIP ; tetrahydroisoxazolopyridinol ; CRR ; concentration&ndash ; response relationship
• 刊名：Biochemical Pharmacology
• 出版年：2016
• 出版时间：1 March 2016
• 年：2016
• 卷：103
• 期：Complete
• 页码：98-108
• 全文大小：1588 K

文摘

GABA_A receptors that contain the α4 and δ subunits are thought to be located extrasynaptically, mediating tonic currents elicited by low concentrations of GABA. These α4βδ receptors are modulated by neurosteroids and certain anesthetics, identifying them as important drug targets in research. However, pharmacological studies on these receptors have often yielded variable results, possibly due to the expression of receptors in different stoichiometries or arrangements.

In this study, we injected different ratios of α4, β2 and δ cRNA into Xenopus oocytes and measured the sensitivity to GABA and DS2 activation of the resulting receptor populations. By creating a matrix of RNA injection ratios from stock RNA concentrations, we were able to compare the changes in pharmacology between injection ratios where the ratio of only one subunit was altered.

We identified two distinct populations of receptors, the first with an EC₅₀ value of approximately 100 nM to GABA, a low Hill slope of approximately 0.3 and substantial direct activation by DS2. The second population had an EC₅₀ value of approximately 1 μM to GABA, a steeper Hill slope of 1 and little direct activation, but substantial potentiation, by DS2. The second population was formed with high α4 ratios and low β2 ratios, but altering the ratio of δ subunit injected had little effect. We propose that receptors with high sensitivity to GABA and direct activation by DS2 are the result of a greater number of β2 subunits being incorporated into the receptor.