Markers for persistent specific expressive language delay in 3-4-year-olds.

BACKGROUND: Identifying 3-4-year-olds who are most at risk of persisting language difficulties, and possibly specific language impairment (SLI), is difficult due to the natural variation of language in young children. In older children, markers for SLI have been identified that differentiate between children with and without SLI. It is not known whether these markers can be used at an earlier age to identify children most at risk of persisting language difficulties. AIMS: To identify possible risk markers of current status that distinguish children who have specific expressive language delay (SELD) from the variation observed in normally developing children at age 3;0-4;0 and 4;0-5;0. To determine the most suitable measure(s) that would predict which children with SELD at age 3;0-4;0 were likely to have persistent expressive language delay (PELD) at age 4;0-5;0. METHODS & PROCEDURES: Forty-seven children with SELD and 47 children with typical language development (TLD) were assessed on language, nonverbal IQ and marker tasks at age 3;0-4;0 (baseline). Ninety-one children were reassessed on the measures one year later (follow-up). At both time points, the marker tasks were compared with a reference standard (Expressive Communication subscale (EC) of the Preschool Language Scale-3 (PLS-3) (UK)) to determine the most useful marker for identifying children with SELD. Possible predictors were examined to determine the most suitable measure(s) that would predict which children with SELD at baseline were likely to have PELD at follow-up. OUTCOME & RESULTS: A modified version of Recalling Sentences, a subtest of the Clinical Evaluation of Language Fundamentals (CELF)-Preschool UK, was the most useful marker for identifying children with SELD at baseline and at follow-up. Thirty-five (76.1%) of the 46 children with SELD at baseline had PELD at follow-up. Performance on the Auditory Comprehension (AC) and EC subscales of the PLS-3 (UK) and on modified Recalling Sentences at age 3;0-4;0 were predictors of PELD at age 4;0-5;0. CONCLUSIONS & IMPLICATIONS: A modified Recalling Sentences task was a good risk marker for SELD at age 3;0-4;0 and SELD at age 4;0-5;0. PLS-3 AC, PLS-3 EC and modified Recalling Sentences at baseline were the best predictors of PELD. The use of modified Recalling Sentences as a predictive marker requires confirmation.

CMIP and ATP2C2 modulate phonological short-term memory in language impairment.

Specific language impairment (SLI) is a common developmental disorder characterized by difficulties in language acquisition despite otherwise normal development and in the absence of any obvious explanatory factors. We performed a high-density screen of SLI1, a region of chromosome 16q that shows highly significant and consistent linkage to nonword repetition, a measure of phonological short-term memory that is commonly impaired in SLI. Using two independent language-impaired samples, one family-based (211 families) and another selected from a population cohort on the basis of extreme language measures (490 cases), we detected association to two genes in the SLI1 region: that encoding c-maf-inducing protein (CMIP, minP = 5.5 x 10(-7) at rs6564903) and that encoding calcium-transporting ATPase, type2C, member2 (ATP2C2, minP = 2.0 x 10(-5) at rs11860694). Regression modeling indicated that each of these loci exerts an independent effect upon nonword repetition ability. Despite the consistent findings in language-impaired samples, investigation in a large unselected cohort (n = 3612) did not detect association. We therefore propose that variants in CMIP and ATP2C2 act to modulate phonological short-term memory primarily in the context of language impairment. As such, this investigation supports the hypothesis that some causes of language impairment are distinct from factors that influence normal language variation. This work therefore implicates CMIP and ATP2C2 in the etiology of SLI and provides molecular evidence for the importance of phonological short-term memory in language acquisition.

Protein interactions involving the gamma2 large cytoplasmic loop of GABA(A) receptors modulate conductance.

Native GABA(A) channels display a single-channel conductance ranging between approximately 10 and 90 pS. Diazepam increases the conductance of some of these native channels but never those of recombinant receptors, unless they are coexpressed with GABARAP. This trafficking protein clusters recombinant receptors in the membrane, suggesting that high-conductance channels arise from receptors that are at locally high concentrations. The amphipathic (MA) helix that is present in the large cytoplasmic loop of every subunit of all ligand-gated ion channels mediates protein-protein interactions. Here we report that when applied to inside-out patches, a peptide mimicking the MA helix of the gamma2 subunit (gamma(381-403)) of the GABA(A) receptor abrogates the potentiating effect of diazepam on both endogenous receptors and recombinant GABA(A) receptors coexpressed with GABARAP, by substantially reducing their conductance. The protein interaction disrupted by the peptide did not involve GABARAP, because a shorter peptide (gamma(386-403)) known to compete with the gamma2-GABARAP interaction did not affect the conductance of recombinant alphabetagamma receptors coexpressed with GABARAP. The requirement for receptor clustering and the fact that the gamma2 MA helix is able to self-associate support a mechanism whereby adjacent GABA(A) receptors interact via their gamma2-subunit MA helices, altering ion permeation through each channel. Alteration of ion-channel function arising from dynamic interactions between ion channels of the same family has not been reported previously and highlights a novel way in which inhibitory neurotransmission in the brain may be differentially modulated.

Differential drug responses on native GABA(A) receptors revealing heterogeneity in extrasynaptic populations in cultured hippocampal neurons.

Hippocampal pyramidal neurons potentially express multiple subtypes of GABA(A) receptors at extrasynaptic locations that could therefore respond to different drugs. We activated extrasynaptic GABA(A) receptors in cultured rat hippocampal pyramidal neurons and measured single-channel currents in order to compare the actions of two drugs that potentially target different GABA(A) receptor subtypes. Despite the possible difference in receptor targets of etomidate and diazepam, the two drugs were similar in their actions on native extrasynaptic GABA(A) receptors. Each drug produced three distinct responses that differed significantly in current magnitude, implying heterogeneous GABA(A) receptor populations. In the majority of patches, drug application increased both the single-channel conductance (>40 pS) and the open probability of the channels. By contrast, in the minority of patches, drug application caused an increase in open probability only. In the third group high-conductance channels were observed upon GABA activation and drug application increased their open probability only. The currents potentiated by etomidate or diazepam were substantially larger in patches displaying high-conductance GABA channels compared to those displaying only low-conductance channels. Factors contributing to the large magnitude of these currents were the long mean open time of high-conductance channels and the presence of multiple channels in these patches. In conclusion, we suggest that the local density of extrasynaptic GABA(A) receptors may influence their single-channel properties and may be an additional regulating factor for tonic inhibition and, importantly, differential drug modulation.

Conductance of recombinant GABA (A) channels is increased in cells co-expressing GABA(A) receptor-associated protein.

High conductance gamma-aminobutyric acid type A (GABA(A)) channels (>40 picosiemens (pS)) have been reported in some studies on GABA(A) channels in situ but not in others, whereas recombinant GABA(A) channels do not appear to display conductances above 40 pS. Furthermore, the conductance of some native GABA(A) channels can be increased by diazepam or pentobarbital, which are effects not reported for expressed GABA(A) channels. GABARAP, a protein associated with native GABA(A) channels, has been reported to cause clustering of GABA(A) receptors and changes in channel kinetics. We have recorded single channel currents activated by GABA in L929 cells expressing alpha(1), beta(1), and gamma(2S) subunits of human GABA(A) receptors. Channel conductance was never higher than 40 pS and was not significantly increased by diazepam or pentobarbital, although open probability was increased. In contrast, in cells expressing the same three subunits together with GABARAP, channel conductance could be significantly higher than 40 pS, and channel conductance was increased by diazepam and pentobarbital. GABARAP caused clustering of receptors in L929 cells, and we suggest that there may be interactions between subunits of clustered GABA(A) receptors that make them open co-operatively to give high conductance "channels." Recombinant channels may require the influence of GABARAP and perhaps other intracellular proteins to adopt a fuller repertoire of properties of native channels.