The contribution of δ subunit-containing GABAA receptors to phasic and tonic conductance changes in cerebellum, thalamus and neocortex

We have made use of the delta subunit-selective allosteric modulator DS2 (4-chloro-N-[2-(2-thienyl)imidazo[1,2-a]pyridine-3-yl benzamide) to assay the contribution of delta-GABAARs to tonic and phasic conductance changes in the cerebellum, thalamus and neocortex. In cerebellar granule cells, an enhancement of the tonic conductance was observed for DS2 and the orthosteric agonist THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol). As expected, DS2 did not alter the properties of GABAA receptor-mediated inhibitory postsynaptic synaptic currents (IPSCs) supporting a purely extrasynaptic role for delta-GABAARs in cerebellar granule cells. DS2 also enhanced the tonic conductance recorded from thalamic relay neurons of the visual thalamus with no alteration in IPSC properties. However, in addition to enhancing the tonic conductance DS2 also slowed the decay of IPSCs recorded from layer II/III neocortical neurons. A slowing of the IPSC decay also occurred in the presence of the voltage-gated sodium channel blocker TTX. Moreover, under conditions of reduced GABA release the ability of DS2 to enhance the tonic conductance was attenuated. These results indicate that delta-GABAARs can be activated following vesicular GABA release onto neocortical neurons and that the actions of DS2 on the tonic conductance may be influenced by the ambient GABA levels present in particular brain regions

Three years of ocean data from a bio-optical profiling float

Ocean color, first measured from space 30 years ago, has provided a revolutionary synoptic view of near-surface fields of phytoplankton pigments. Since 1979, a number of ocean color satellite missions have provided coverage of phytoplankton biomass and other biogeochemical variables on scales of days to years and of kilometers to ocean basin. Because of the nature of visible light and its interaction with absorbing and scattering materials in the ocean and atmosphere, these measurements are biased toward nearsurface waters and are obscured by clouds. As a consequence, ocean color satellites miss significant fractions of phytoplankton biomass, marine primary productivity, and particle flux that occur at depths beyond their sensing range. They also miss phytoplankton blooms and other events that occur during periods of extended cloud cover

GABA Transporter Deficiency Causes Tremor, Ataxia, Nervousness, and Increased GABA-Induced Tonic Conductance in Cerebellum

GABA transporter subtype 1 (GAT1) knock-out (KO) mice display normal reproduction and life span but have reduced body weight (female, -10%; male, -20%) and higher body temperature fluctuations in the 0.2-1.5/h frequency range. Mouse GAT1 (mGAT1) KO mice exhibit motor disorders, including gait abnormality, constant 25-32 Hz tremor, which is aggravated by flunitrazepam, reduced rotarod performance, and reduced locomotor activity in the home cage. Open-field tests show delayed exploratory activity, reduced rearing, and reduced visits to the central area, with no change in the total distance traveled. The mGAT1 KO mice display no difference in acoustic startle response but exhibit a deficiency in prepulse inhibition. These open-field and prepulse inhibition results suggest that the mGAT1 KO mice display mild anxiety or nervousness. The compromised GABA uptake in mGAT1 KO mice results in an increased GABA_A receptor-mediated tonic conductance in both cerebellar granule and Purkinje cells. The reduced rate of GABA clearance from the synaptic cleft is probably responsible for the slower decay of spontaneous IPSCs in cerebellar granule cells. There is little or no compensatory change in other proteins or structures related to GABA transmission in the mGAT1 KO mice, including GAT1-independent GABA uptake, number of GABAergic interneurons, and GABA_A-, vesicular GABA transporter-, GAD65-, and GAT3-immunoreactive structures in cerebellum or hippocampus. Therefore, the excessive extracellular GABA present in mGAT1 KO mice results in behaviors that partially phenocopy the clinical side effects of tiagabine, suggesting that these side effects are inherent to a therapeutic strategy that targets the widely expressed GAT1 transporter system

Signaling across the synapse: a role for Wnt and Dishevelled in presynaptic assembly and neurotransmitter release

Proper dialogue between presynaptic neurons and their targets is essential for correct synaptic assembly and function. At central synapses, Wnt proteins function as retrograde signals to regulate axon remodeling and the accumulation of presynaptic proteins. Loss of Wnt7a function leads to defects in the localization of presynaptic markers and in the morphology of the presynaptic axons. We show that loss of function of Dishevelled-1 (Dvl1) mimics and enhances the Wnt7a phenotype in the cerebellum. Although active zones appear normal, electrophysiological recordings in cerebellar slices from Wnt7a/Dvl1 double mutant mice reveal a defect in neurotransmitter release at mossy fiber–granule cell synapses. Deficiency in Dvl1 decreases, whereas exposure to Wnt increases, synaptic vesicle recycling in mossy fibers. Dvl increases the number of Bassoon clusters, and like other components of the Wnt pathway, it localizes to synaptic sites. These findings demonstrate that Wnts signal across the synapse on Dvl-expressing presynaptic terminals to regulate synaptic assembly and suggest a potential novel function for Wnts in neurotransmitter release

Combining mGRASP and Optogenetics Enables High-Resolution Functional Mapping of Descending Cortical Projections

We have applied optogenetics and mGRASP, a light microscopy technique that labels synaptic contacts, to map the number and strength of defined corticocollicular (CC) connections. Using mGRASP, we show that CC projections form small, medium, and large synapses, and both the number and the distribution of synapse size vary among the IC regions. Using optogenetics, we show that low-frequency stimulation of CC axons expressing channelrhodopsin produces prolonged elevations of the CC miniature EPSC (mEPSC) rate. Functional analysis of CC mEPSCs reveals small-, medium-, and large-amplitude events that mirror the synaptic distributions observed with mGRASP. Our results reveal that descending ipsilateral projections dominate CC feedback via an increased number of large synaptic contacts, especially onto the soma of IC neurons. This study highlights the feasibility of combining microscopy (i.e., mGRASP) and optogenetics to reveal synaptic weighting of defined projections at the level of single neurons, enabling functional connectomic mapping in diverse neural circuits

Tectal-derived interneurons contribute to phasic and tonic inhibition in the visual thalamus

The release of GABA from local interneurons in the dorsal lateral geniculate nucleus (dLGN-INs) provides inhibitory control during visual processing within the thalamus. It is commonly assumed that this important class of interneurons originates from within the thalamic complex, but we now show that during early postnatal development Sox14/Otx2-expressing precursor cells migrate from the dorsal midbrain to generate dLGN-INs. The unexpected extra-diencephalic origin of dLGN-INs sets them apart from GABAergic neurons of the reticular thalamic nucleus. Using optogenetics we show that at increased firing rates tectal-derived dLGN-INs generate a powerful form of tonic inhibition that regulates the gain of thalamic relay neurons through recruitment of extrasynaptic high-affinity GABA(A) receptors. Therefore, by revising the conventional view of thalamic interneuron ontogeny we demonstrate how a previously unappreciated mesencephalic population controls thalamic relay neuron excitability.Peer reviewe

Synaptic mechanisms in experience-dependent processes responsible for the development of central neural maps of visual space

Experience-dependent synaptic plasticity has been demonstrated in the binocular visual system of the aquatic frog Xenopus laevis. This thesis offers further support for the role of correlated patterns of neural activity in this synaptic plasticity and demonstrates, for the first time, that N-methyl-D-aspartate (NMDA)-type glutamate receptors contribute to synaptic transmission in the optic tectum of Xenopus. Electrophysiological mapping techniques assessed the capacity for synaptic reorganisations to occur in the binocular visual system of Xenopus. The intertectal system, a series of connections that mediates binocular vision in lower vertebrates, reorganised its pattern of synaptic connections in response to large changes in the orientation of both eyes. Stroboscopic illumination was shown to interfere with correlated patterns of neural activity in the tectum, but the development of the retinotectal projection was not affected by this procedure. However, the intertectal systems capacity to modify its pattern of synaptic connections, in response to changes in eye position, was disturbed by this visual environment. Next, in vivo and in vitro preparations of the optic tectum were developed to monitor pre and postsynaptic activity in the visual system of Xenopus. It was established that mono-synaptic transmission between retinal fibres and tectal neurons was mediated by non-NMDA type glutamate receptors. However, when the optic tract of the adult animal was electrically stimulated the late poly-synaptic (U2) component of the evoked response was reduced by ˜30% in the presence of 50μM AP5. This NMDA mediated component of the late U2 response was at its most significant 1 month after metamorphosis, was regulated by visual experience, and was not present in another species of frog, Rana pipiens. Finally, a thin slice preparation of the optic tectum was developed which enabled tight-seal whole cell recordings to be made from identified tectal neurons