Directional summation in non-direction selective retinal ganglion cells.

Retinal ganglion cells receive inputs from multiple bipolar cells which must be integrated before a decision to fire is made. Theoretical studies have provided clues about how this integration is accomplished but have not directly determined the rules regulating summation of closely timed inputs along single or multiple dendrites. Here we have examined dendritic summation of multiple inputs along On ganglion cell dendrites in whole mount rat retina. We activated inputs at targeted locations by uncaging glutamate sequentially to generate apparent motion along On ganglion cell dendrites in whole mount retina. Summation was directional and dependent13 on input sequence. Input moving away from the soma (centrifugal) resulted in supralinear summation, while activation sequences moving toward the soma (centripetal) were linear. Enhanced summation for centrifugal activation was robust as it was also observed in cultured retinal ganglion cells. This directional summation was dependent on hyperpolarization activated cyclic nucleotide-gated (HCN) channels as blockade with ZD7288 eliminated directionality. A computational model confirms that activation of HCN channels can override a preference for centripetal summation expected from cell anatomy. This type of direction selectivity could play a role in coding movement similar to the axial selectivity seen in locust ganglion cells which detect looming stimuli. More generally, these results suggest that non-directional retinal ganglion cells can discriminate between input sequences independent of the retina network.

Calcium dynamics encode the magnitude of a graded memory underlying sensorimotor adaptation.

The role of Ca(2+) in the induction of neural correlates of memory has frequently been described in binary terms despite the fact that many forms of memory are graded in their strength and/or persistence. We find that Ca(2+) dynamics encode the magnitude of sensorimotor adaptation of the electromotor output in a weakly electric fish. The neural correlate of this memory is a synaptically induced Ca(2+)-dependent enhancement of intrinsic excitability of neurons responsible for setting the electromotor output. Changes in Ca(2+) during induction accurately predict the magnitude of this graded memory over a wide range of stimuli. Thus despite operating over a range from seconds to tens of minutes, the encoding of graded memory can be mediated by a relatively simple cellular mechanism.

Protein kinase A regulates calcium permeability of NMDA receptors.

Calcium (Ca2+) influx through NMDA receptors (NMDARs) is essential for synaptogenesis, experience-dependent synaptic remodeling and plasticity. The NMDAR-mediated rise in postsynaptic Ca2+ activates a network of kinases and phosphatases that promote persistent changes in synaptic strength, such as long-term potentiation (LTP). Here we show that the Ca2+ permeability of neuronal NMDARs is under the control of the cyclic AMP-protein kinase A (cAMP-PKA) signaling cascade. PKA blockers reduced the relative fractional Ca2+ influx through NMDARs as determined by reversal potential shift analysis and by a combination of electrical recording and Ca2+ influx measurements in rat hippocampal neurons in culture and hippocampal slices from mice. In slices, PKA blockers markedly inhibited NMDAR-mediated Ca2+ rises in activated dendritic spines, with no significant effect on synaptic current. Consistent with this, PKA blockers depressed the early phase of NMDAR-dependent LTP at hippocampal Schaffer collateral-CA1 (Sch-CA1) synapses. Our data link PKA-dependent synaptic plasticity to Ca2+ signaling in spines and thus provide a new mechanism whereby PKA regulates the induction of LTP.

Asymmetric rostro-caudal inhibition in the primary olfactory cortex.

The importance of intracortical inhibitory circuits in setting the feature-selective spatial organization of primary sensory cortices remains controversial. To address this issue, we examined the strength of interneuron-to-pyramidal cell connections across the rat anterior piriform cortex (aPC) and found a pronounced gradient of increasing pyramidal cell inhibition along the aPC rostro-caudal axis. This functional heterogeneity could govern aPC spatial activation in response to varying odor identities and features.

Recruiting extrasynaptic NMDA receptors augments synaptic signaling.

N-Methyl-d-aspartate receptor (NMDAR) activation may promote cell survival or initiate cell death, with the outcome dependent on whether synaptic or extrasynaptic receptors are activated. Similarly, this differential activation has been proposed to govern the direction of plasticity. However, the physiological parameters necessary to activate extrasynaptic NMDARs in brain slices remain unknown. Using the irreversible use-dependent NMDAR antagonist MK-801 to isolate extrasynaptic NMDARs, we have tested the ability of short-stimulation trains from 5 to 400 Hz to activate these receptors on CA1 hippocampal slice pyramidal neurons. Frequencies as low as 25 Hz engage extrasynaptic NMDARs, with maximal activation at frequencies between 100 and 200 Hz. Since similar bursts of synaptic input occur during exploratory behavior in rats, our results demonstrate that "extrasynaptic" NMDARs regularly participate in synaptic transmission. Further, 175-Hz-stimulation trains activate all available synaptic and extrasynaptic dendritic NMDARs, suggesting these NMDARs act as synaptic receptors as needed, transiently increasing synaptic strength. Thus extrasynaptic NMDARs play a vital role in synaptic physiology, calling into question their status as "extrasynaptic."

Extrasynaptic and synaptic NMDA receptors form stable and uniform pools in rat hippocampal slices.

N-methyl-d-aspartate receptor (NMDAR) activation can trigger both long- and short-term plasticity, promote cell survival, and initiate cell death. A number of studies suggest that the consequences of NMDAR activation can vary widely depending on whether synaptic or extrasynaptic receptors are activated. Here we have examined the spatial distribution of NMDARs of CA1 pyramidal neurons in acutely dissected hippocampal slices. Using a physiological definition of extrasynaptic receptors as those not accessible to single release events, we find that extrasynaptic NMDARs comprise a substantial proportion of the dendritic NMDAR pool (36%). This pool of extrasynaptic NMDARs is stable and does not shuttle into the synaptic receptor pool, as we observe no recovery of synaptic current after MK-801 synaptic blockade and washout. The subunit composition of synaptic and extrasynaptic NMDA receptor pools is similar at 3 weeks of age, with NR2B subunits present in both compartments. NR2B receptors are not enriched in the extrasynaptic compartment. Our data suggest that any role played by extrasynaptic NMDARs in synaptic transmission is dictated by their subcellular location rather than their subunit composition or mobility.

Chemical two-photon uncaging.

INTRODUCTIONChemical two-photon uncaging is useful for a wide range of applications, including both mapping of receptor location and localized photostimulation of neurons via activation of excitatory glutamate receptors. Experimental preparations could include brain slices, cultured neurons, and, among other possibilities, whole brains in vivo. This protocol documents the utility of chemical two-photon uncaging in examining glutamate receptors of pyramidal neurons in hippocampal slices.

Synaptic kainate currents reset interneuron firing phase.

Hippocampal interneuron activity has been linked to epileptogenesis, seizures and the oscillatory synaptic activity detected in behaving rats. Interneurons fire at specific times in the rhythmic cycles that comprise these oscillations; however, the mechanisms controlling these firing patterns remain unclear. We have examined the role of synaptic input in modulating the firing of spontaneously active rat hippocampal interneurons. We find that synaptic glutamate receptor currents of 20-30 pA increase instantaneous firing frequency and reset the phase of spontaneously firing CA1 stratum oriens interneurons. Kainate receptor (KAR)-mediated currents are particularly effective at producing this phase reset, while AMPA receptor currents are relatively ineffective. The efficacy of KAR-mediated currents is probably due to their 3-fold longer decay. Given the small amplitude of the currents needed for this phase reset, coincident activation of only a few KAR-containing synapses could synchronize firing in groups of interneurons. These data suggest that KARs are potent modulators of circuit behaviour and their activation alters hippocampal interneuron output.

Variable kainate receptor distributions of oriens interneurons.

Interneuron kainate receptor (KAR) activation regulates normal network activity and modulates cell excitability. As a result, determining the subcellular distribution of KARs in a cell-specific manner is a necessary step toward understanding their role in network function. We have functionally mapped synaptic and extrasynaptic dendritic KARs on hippocampal oriens interneurons using local photolysis of caged glutamate. We find that the majority of trilaminar and oriens lacunosum-moleculare (O-LM) cells have uniform and continuous current densities along the lengths of their dendrites. However, there is a subpopulation of interneurons that have no KAR currents or currents exclusively at "hot spots" on the soma and dendrites. Finally, bistratified cells have KAR currents on all dendrites except those extending into the stratum radiatum. Thus KARs are functionally distributed in a cell-specific and cell-independent manner that may reflect the physiologically distinct roles they play in the hippocampal network.