Where have all the Orais gone? Commentary on "Orai1 channels are essential for amplification of glutamate-evoked Ca²⁺ signals in dendritic spines to regulate working and associative memory"

Orai1 channels were reported as critical contributors to the Ca2+ signal in hippocampal neurons underlying synaptic plasticity associated with learning and memory. We discuss the results in view of conflicting other reports that stressed the roles of Orai2 channels but failed to detect functions of Orai1 channels in these neurons

Patch clamp techniques used for studying synaptic transmission in slices of mammalian brain.

Procedures are described for recording postsynaptic currents from neurones in slices of rat brain using patch clamp techniques. The method involves cutting brain slices (120-300, µm thick) with a vibrating microtome followed by localization of cell somata, which can be clearly seen with Nomarski differential interference contrast optics in the light microscope. Tissue covering the identified cell is then removed mechanically and standard patch clamp techniques are applied. Using these methods, spontaneously occurring and stimulus-evoked inhibitory postsynaptic currents (IPSCs) were recorded from neurones in rat hippocampus at greatly improved resolution. In the presence of tetrodotoxin, to block presynaptic action potentials, spontaneous IPSCs seldom exceeded 25 pA. Evoked IPSCs elicited by constant electrical stimulation of a presynaptic neurone were larger and fluctuated in their amplitudes. Single-channel currents, activated by the putative inhibitory transmitter γ-aminobutyric acid (GABA), had a size of about 1 pA. The number of postsynaptic channels activated by a packet of inhibitory transmitter is probably not more than thirty, nearly two orders of magnitude smaller than previously reported estimates for CNS synapses. This might reflect matching of synaptic efficacy to the high input resistance of hippocampal neurones and could be a requirement for fine tuning of inhibition

Voltage sensitivity of NMDA-receptor mediated postsynaptic currents.

Patch-clamp techniques were used to record pharmacologically-isolated N-methyl-D-aspartate-mediated excitatory postsynaptic currents (NMDA-EPSCs) from dentate granule cells in thin rat hippocampal slices. Membrane voltage modulated these EPSCs in two ways. Firstly, depolarization from resting potential enhanced EPSC amplitudes, as expected for a voltage-dependent block by Mg2+ of synaptically activated NMDA receptor channels. Secondly, depolarization markedly prolonged the time course of decay of NMDA-EPSCs in normal and low extracellular Mg2+. Both mechanisms were complementary in establishing a strong dependence between membrane potential and the amount of charge, namely Ca2+, transferred through synaptically activated NMDA receptor channels, that presumably underlies induction of long-term potentiation in the hippocampus

A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system.

(1) A preparation is described which allows patch clamp recordings to be made on mammalian central nervous system (CNS) neurones in situ. (2) A vibrating tissue slicer was used to cut thin slices in which individual neurones could be identified visually. Localized cleaning of cell somata with physiological saline freed the cell membrane, allowing the formation of a high resistance seal between the membrane and the patch pipette. (3) The various configurations of the patch clamp technique were used to demonstrate recording of membrane potential, whole cell currents and single channel currents from neurones and isolated patches. (4) The patch clamp technique was used to record from neurones filled with fluorescent dyes. Staining was achieved by filling cells during recording or by previous retrograde labelling. (5) Thin slice cleaning and patch clamp techniques were shown to be applicable to the spinal cord and almost any brain region and to various species. These techniques are also applicable to animals of a wide variety of postnatal ages, from newborn to adult

Vanishing spin alignment : experimental indication of triaxial 28Si+28Si\bf ^{28}Si + {^{28}Si} nuclear molecule

Fragment-fragment-$\gamma$ coincidences have been measured for $\rm ^{28}Si + {^{28}Si}$ at an energy corresponding to the population of a conjectured resonance in $^{56}$Ni. Fragment angular distributions as well as $\gamma$-ray angular correlations indicate that the spin orientations of the outgoing fragments are perpendicular to the orbital angular momentum. This differs from the $\rm ^{24}Mg+{^{24}Mg}$ and the $\rm ^{12}C+{^{12}C}$ resonances, and suggests two oblate $\rm ^{28}Si$ nuclei interacting in an equator-to-equator molecular configuration.Comment: 14 pages standard REVTeX file, 3 ps Figures -- Accepted for publication in Physical Review C (Rapid Communication