Article ID Journal Published Year Pages File Type
6273583 Neuroscience 2014 11 Pages PDF
Abstract

•The kinetic of Cl− transport was quantified in rat and human neocortical neurons.•The KCC2 blocker VU0240551 decreased of Cl− transport in rat and human neurons.•In about two-thirds of human neurons KCC2 are reduced by about 70% vs. the others.•The transport rate by the anion exchanger was indistinguishable in rat and human neurons.

The normal function of GABAA receptor-mediated inhibition is governed by several factors, including release of GABA, subunit composition and density of the receptors and in particular by the appropriate ionic gradient. In the human epileptogenic neocortex an impaired chloride (Cl−) gradient has been proposed, due to decreases of potassium-coupled chloride transport (KCC2) and voltage-gated Cl− channels (ClC). Regarding sodium- and potassium-coupled Cl− transport (NKCC1) both up- and downregulations have been proposed.We investigated changes of Cl− homeostasis of human and rat neocortical neurons (layer 2/3) with intracellular recordings and iontophoretic Cl− loading employing selective compounds. After cessation of iontophoresis, the IPSPA amplitudes of rat neurons recovered with a time constant (τrec) of 6.5 s (n = 21). In human neurons, τrec averaged 17.8 s (n = 36; 23 resections). Application of the novel KCC2 blocker VU0240551 (1 μM) caused in rat neurons a reversible prolongation of τrec from 5.7 to 8.1 s (n = 11), corresponding to a VU0240551-sensitive Cl− transport rate (1/Δτrec) of 0.0504 s−1. In human neurons, τrec increased on application of 1 μM VU0240551, on average from 15.1 to 20.3 s (n = 17). The human neurons comprised two subgroups with different τrec when segregated according to a border given by the mean + 2 s.d. of rat neurons. In one group, τrec averaged 8.7 s (n = 6) and reversibly increased to 14.6 s in the presence of 1 μM VU0240551, corresponding to a Cl− transport rate of 0.0504 s−1. The other group had an average τrec of 18.5 s which increased in the presence of 1 μM VU0240551 to 23.3 s (n = 11), indicating a much smaller rate (0.0151 s−1). Addition of DIDS, a presumed blocker of anion exchanger (AE), increased the τrec of rat neurons from 7.5 to 8.8 s (n = 6) corresponding to a DIDS-sensitive rate of 0.0185 s−1. In human neurons, DIDS increased τrec from 23.3 to 50.7 s (n = 7), corresponding to a DIDS-sensitive rate of 0.0200 s−1.These data suggest a greatly reduced KCC2-mediated transport rate in most of the human neurons. The two subgroups observed in human tissue indicate a considerable variability of Cl− transport within a given tissue from almost normal to greatly impeded, predominated by a decline of KCC2 whereas AE is unaltered.

Related Topics
Life Sciences Neuroscience Neuroscience (General)
Authors
, , , ,