High and low GABA sensitivity alpha 4 beta 2 delta GABA(A) receptors are expressed in Xenopus laevis oocytes with divergent stoichiometries

作者：Hartiadi Leonny Y; Ahring Philip K; Chebib Mary; Absalom Nathan L^*

来源：Biochemical Pharmacology, 2016, 103: 98-108.

DOI：10.1016/j.bcp.2015.12.021

• Cys-loop receptor
• GABA(A) receptor
• Delta subunit
• DS2
• Stoichiometry

GABA(A) receptors that contain the alpha 4 and delta subunits are thought to be located extrasynaptically, mediating tonic currents elicited by low concentrations of GABA. These alpha 4 beta delta 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 alpha 4, beta 2 and delta cRNA into Xenopus oocytes and measured the sensitivity to GAB(A) 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 03 and substantial direct activation by DS2. The second population had an EC50 value of approximately mu M to GABA, a steeper Hill slope of 1 and little direct activation, but substantial potentiation, by DS2. The second population was formed with high alpha 4 ratios and low beta 2 ratios, but altering the ratio of delta 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 beta 2 subunits being incorporated into the receptor.

• 出版日期2016-3-1