1 Effects of apamin on rat sympathetic neurones were investigated by means of intracellular and extracellular recording. 2 Apamin (50 nM) significantly shortened the after-hyperpolarization (AH) following the spike evoked by current injection and slightly decreased its peak amplitude without affecting the time course of the spike. 3 The AH following the synaptically-evoked spike was also blocked by apamin. This effect was doseand time-dependent (ID50 estimated by extracellular recording approximately 15 nM, 20 min after application) and poorly reversible. Transmission of a single volley was not affected by 50 nM apamin. 4 Though a long depolarizing current caused one or two spikes in the cell, greater repetitive firing was observed in the presence of apamin. Spontaneous repetitive firing, however, was not observed except for anodal-break spikes. Resting potential and input membrane resistance were essentially unchanged by apamin. 5 The maximum rate of rise of the Ca spike was not decreased by 50 nM apamin but the duration of the spike was lengthened by 60%. The AH following the Ca spike was also blocked by apamin. 6 These results suggest that apamin suppressed the slow AH without any inhibition of the Ca flux into the cell and is useful as a blocker of GK(Ca) in the rat sympathetic neurone.

Macrophages within the murine tunica muscularis were isolated and cultured for physiological studies. Following dispersion, macrophages were identified by phagocytotic activity of fluorescein isothiocyanate (FITC)-dextran. Immediately following isolation, macrophages were rounded and possessed fluorescent granula but developed a ramified shape after 3-4 days in culture. Resident and cultured macrophages were immunopositive for F4/80 and I-Ad/I-Ed. Greater than 90% of F4/80 positive cultured cells were FITC-dextran positive. Macrophages had resting membrane potentials (RMP) of -33.3 +/- 1.5 mV after 1 day in culture, which increased to -53.9 +/- 4.4 mV after 3-4 days. The change in RMP was associated with the development of an inward rectifying K+ current, and a decrease in a voltage-dependent, inactivating outward current. After 3-4 days in culture the inflammatory mediated substances adenosine triphosphate (ATP), platelet-activating factor and bacterial lipopolysaccharide induced increases in cytoplasmic Ca2+ ([Ca2+]i). Forskolin suppressed the ATP-induced increase in [Ca2+]i. Macrophages exhibited oxidative bursts, measured by oxidation of dihydrorhodamine-123 to rhodamine-123. Oxidative bursts coincided with a reduction in intracellular pH. Macrophages expressed a proton conductance that may participate in pH maintenance during reactive oxygen production. These results suggest that resident macrophages in the intestine may play a role in the immunological protection of the gut.

We have developed a spherical aberration corrected transmission electron microscopy (Cs-corrected TEM) technique that allows us to obtain clearer images in real space than ever before. We applied this technique to titanium oxide, in which light elements such as oxygen are difficult to observe using TEM because of its small cross section and electronic damage. In the present study, we successfully observed oxygen atoms in rutile TiO2. In addition, this direct observation of oxygen atoms enabled us to study the Magnéli structure (TinO2n−1), which is caused by oxygen vacancies. These vacancies caused an atomic relaxation of the titanium and oxygen atoms. The relaxed atoms formed a characteristic shear structure of rutile titanium dioxide phase. This shear structure of the Magnéli structure (TinO2n−1) was visualized with a spatial resolution of 0.119 nm. At the same time, the selected area diffraction (SAD) pattern of the defect structure was obtained. Additional spots were shown inside the rutile [110] spot. We made structural models of the shear structure and simulated the diffraction pattern and images using a multi-slice simulation. Additional spots in the simulated diffraction patterns accurately reconstructed the experimental data. We also considered the possibility of the real-space analysis of local structures using spherical aberration corrected transmission electron microscopy.

Effects of ryanodine on sympathetic neurones of the rat superior cervical ganglion were investigated by means of intracellular recording. Ryanodine (1 microM) significantly shortened the after-hyperpolarization (AH) following the spike evoked by current injection or pre-ganglionic stimulation without affecting the configuration of the spikes. The shortening of AH caused by ryanodine was dose-dependent at concentrations between 0.1 and 1 microM and was slowly recovered by washing the tissue over 1 h. A partial inhibition of the apamin-sensitive slow component of AH was the maximal effect obtained at 1 microM. Although the input membrane resistance was not changed, ryanodine evoked repetitive discharges at long intervals in response to long depolarizing current pulses applied across the cell membrane. Ryanodine (5 microM) did not depress the Ca-spike but shortened the following AH in a lesser degree than that following the normal spike. Spontaneous small fluctuations of the resting membrane potential were occasionally observed under normal conditions. They were facilitated by caffeine and abolished by ryanodine. Caffeine also enhanced the slow component of the AH but did not affect it in the presence of ryanodine. These results suggest that ryanodine inhibits Ca release from intracellular store sites. The released Ca may contribute to generating the long-lasting AH and to regulating the excitability of rat sympathetic neurones.

1 The effects of tetraethylammonium (TEA) and 4-aminopyridine (4-AP) on

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