The synapse-bearing nerve terminals of the opener muscle of the crayfish Procambarus were reconstructed using electron micrographs of regions which had

Using the whole-cell recording technique, we have examined the slow Ca(2+)-activated afterhyperpolarization (AHP) and its underlying current (IAHP) in hippocampal CA1 neurones of brain slices obtained from mature rats. Specifically we have studied the effects of the anion component of various K+ salts commonly used to make the pipette filling solution that dialyses neurones during whole-cell recordings. Among the K+ salts examined which included potassium methylsulfate, potassium methanesulfonate, potassium gluconate, potassium chloride, potassium citrate and potassium glutamate, stable AHPs/IAHP and strong spike firing adaptation could only be observed in neurones recorded with the patch pipette solution containing potassium methylsulfate. These AHPs and firing patterns closely mimicked those recorded with sharp electrodes. Similarly, the sustained component of voltage-activated Ca2+ currents was more stable in neurones dialysed with cesium methanesulfonate than in those dialysed with cesium gluconate or cesium chloride. Although the mechanisms underlying the interaction(s) between internally applied anions and ionic channels need further investigation, the present experiments illustrate that in mammalian brain neurones at 33 degrees C, the Ca(2+)-activated IAHP is dramatically altered by internal anions. We suggest that among anions commonly used in electrode filling solutions for whole-cell recordings, methylsulfate is the least disruptive to intracellular structures or Ca2+ homeostasis and permits stable whole-cell recording of the IAHP and Ca2+ currents in mammalian CNS neurones.

Gap junctions (gjs) are increasingly recognized as playing a significant role in seizures. We demonstrate that different types of gap junctional blocking agents reduce the duration of evoked seizure-like primary afterdischarges (PADs) in the rat in vitro CA1 hippocampal pyramidal region, following repetitive tetanization of the Schaffer collaterals. Intracellular acidosis, which is known to block gap junctional communication, decreased the PADs, whereas alkalinization increased the PADs. Cellular excitability was not significantly depressed as determined by input/output relations recorded before and during perfusion of the gj blockers blockers carbenoxolone and sodium propionate. There was a small decrease following 1-octanol perfusion and a large decrease following NH4Cl application. Carbenoxolone diminished PAD duration, but increased neuronal excitability in whole-cell recordings. After robust PADs were established, the expression of several gj proteins including connexins (Cxs) 26, 32, 36, and 43, as measured by Western blotting, was unchanged, although the level of nonphosphorylated Cx43 was decreased. Our data support the concept that blocking gap junctional communication is an anticonvulsant mechanism.