Rett syndrome (RTT) is a rare, X-linked neurodevelopmental disorder typically affecting females, resulting in a range of symptoms including autistic features, intellectual impairment, motor deterioration, and autonomic abnormalities. RTT is primarily caused by the genetic mutation of the Mecp2 gene. Initially considered a neuronal disease, recent research shows that glial dysfunction contributes to the RTT disease phenotype. In the following manuscript, we review the evidence regarding glial dysfunction and its effects on disease etiology.

Significance: Recent genome-wide association studies suggest a strong linkage between psychiatric disorders, especially bipolar disorder (BPD) and schizophrenia, and ion channels, including G-protein–activated K ⁺ channels (GIRK); however, there are no clear functional links. Lithium is a prominent antibipolar treatment. We report a dual regulation of GIRK channels by therapeutic doses of lithium in neurons. We reproduced these regulations in Xenopus oocytes, identified the molecular mechanism (a dual regulation of G proteins via actions on Gα subunit), and verified the action of Li ⁺ on Gα–Gβγ interaction by direct biochemical studies. The discovery of a significant regulation of an ion channel by therapeutic doses of lithium may help linking between neuronal excitability and mechanisms of BPD.

Rett syndrome (RTT) is an X-linked neurodevelopmental disorder usually caused by mutations in methyl-CpG-binding protein 2 (MeCP2). RTT is typified by apparently normal development until 6–18 mo of age, when motor and communicative skills regress and hand stereotypies, autonomic symptoms, and seizures present. Restoration of MeCP2 function selectively to astrocytes reversed several deficits in a murine model of RTT, but the mechanism of this rescue is unknown. Astrocytes carry out many essential functions required for normal brain functioning, including extracellular K⁺buffering. Kir4.1, an inwardly rectifying K⁺channel, is largely responsible for the channel-mediated K⁺regulation by astrocytes. Loss-of-function mutations in Kir4.1 in human patients result in a severe neurodevelopmental disorder termed EAST or SESAME syndrome. Here, we evaluated astrocytic Kir4.1 expression in a murine model of Rett syndrome. We demonstrate by chromatin immunoprecipitation analysis that Kir4.1 is a direct molecular target of MeCP2. Astrocytes fromMecp2-deficient mice express significantly less Kir4.1 mRNA and protein, which translates into a >50% deficiency in Ba²⁺-sensitive Kir4.1-mediated currents, and impaired extracellular potassium dynamics. By examining astrocytes in isolation, we demonstrate that loss of Kir4.1 is cell autonomous. Assessment through postnatal development revealed that Kir4.1 expression inMecp2-deficient animals never reaches adult, wild-type levels, consistent with a neurodevelopmental disorder. These are the first data implicating a direct MeCP2 molecular target in astrocytes and provide novel mechanistic insight explaining a potential mechanism by which astrocytic dysfunction may contribute to RTT.

G protein-gated K⁺ channels (GIRK; Kir3), activated by Gβγ subunits derived from G_i/o proteins, regulate heartbeat and neuronal excitability and plasticity. Both neurotransmitter-evoked (I_evoked) and neurotransmitter-independent basal (I_basal) GIRK activities are physiologically important, but mechanisms of I_basal and its relation to I_evoked are unclear. We have previously shown for heterologously expressed neuronal GIRK1/2, and now show for native GIRK in hippocampal neurons, that I_basal and I_evoked are interrelated: the extent of activation by neurotransmitter (activation index, R_a) is inversely related to I_basal. To unveil the underlying mechanisms, we have developed a quantitative model of GIRK1/2 function. We characterized single-channel and macroscopic GIRK1/2 currents, and surface densities of GIRK1/2 and Gβγ expressed in Xenopus oocytes. Based on experimental results, we constructed a mathematical model of GIRK1/2 activity under steady-state conditions before and after activation by neurotransmitter. Our model accurately recapitulates I_basal and I_evoked in Xenopus oocytes, HEK293 cells and hippocampal neurons; correctly predicts the dose-dependent activation of GIRK1/2 by coexpressed Gβγ and fully accounts for the inverse I_basal-R_a correlation. Modeling indicates that, under all conditions and at different channel expression levels, between 3 and 4 Gβγ dimers are available for each GIRK1/2 channel. In contrast, available Gα_i/o decreases from ~2 to less than one Gα per channel as GIRK1/2's density increases. The persistent Gβγ/channel (but not Gα/channel) ratio support a strong association of GIRK1/2 with Gβγ, consistent with recruitment to the cell surface of Gβγ, but not Gα, by GIRK1/2. Our analysis suggests a maximal stoichiometry of 4 Gβγ but only 2 Gα_i/o per one GIRK1/2 channel. The unique, unequal association of GIRK1/2 with G protein subunits, and the cooperative nature of GIRK gating by Gβγ, underlie the complex pattern of basal and agonist-evoked activities and allow GIRK1/2 to act as a sensitive bidirectional detector of both Gβγ and Gα. Many neurotransmitters and hormones inhibit the electric activity of excitable cells (such as cardiac cells and neurons) by activating a K⁺ channel, GIRK (G protein-gated Inwardly Rectifying K⁺ channel). GIRK channels also possess constitutive “basal” activity which contributes to regulation of neuronal and cardiac excitability and certain disorders, but the mechanism of this activity and its interrelation with the neurotransmitter-evoked activity are poorly understood. In this work we show that key features of basal and neurotransmitter-evoked activities are similar in cultured hippocampal neurons and in two model systems (mammalian HEK293 cells and Xenopus oocytes). Using experimental data of the neuronal GIRK1/2 channel function upon changes in GIRK and G protein concentrations, we constructed a mathematical model that quantitatively accounts for basal and evoked activity, and for the inverse correlation between the two. Our analysis suggests a novel and unexpected mechanism of interaction of GIRK1/2 with the G protein subunits, where the tetrameric GIRK channel can assemble with 4 molecules of the Gβγ subunits but only 2 molecules of Gα. GIRK is a prototypical effector of Gβγ, and the unequal stoichiometry of interaction with G protein subunits may have general implications for G protein signaling.