Synaptic vesicle glycoprotein 2A modulates vesicular release and calcium channel function at peripheral sympathetic synapses

Synaptic vesicle glycoprotein (SV)2A is a transmembrane protein found in secretory vesicles and is critical for Ca2+-dependent exocytosis in central neurons, although its mechanism of action remains uncertain. Previous studies have proposed, variously, a role of SV2 in the maintenance and formation of the readily releasable pool (RRP) or in the regulation of Ca2+ responsiveness of primed vesicles. Such previous studies have typically used genetic approaches to ablate SV2 levels; here, we used a strategy involving small interference RNA (siRNA) injection to knockdown solely presynaptic SV2A levels in rat superior cervical ganglion (SCG) neuron synapses. Moreover, we investigated the effects of SV2A knockdown on voltage-dependent Ca2+ channel (VDCC) function in SCG neurons. Thus, we extended the studies of SV2A mechanisms by investigating the effects on vesicular transmitter release and VDCC function in peripheral sympathetic neurons. We first demonstrated an siRNA-mediated SV2A knockdown. We showed that this SV2A knockdown markedly affected presynaptic function, causing an attenuated RRP size, increased paired-pulse depression and delayed RRP recovery after stimulus-dependent depletion. We further demonstrated that the SV2A–siRNA-mediated effects on vesicular release were accompanied by a reduction in VDCC current density in isolated SCG neurons. Together, our data showed that SV2A is required for correct transmitter release at sympathetic neurons. Mechanistically, we demonstrated that presynaptic SV2A: (i) acted to direct normal synaptic transmission by maintaining RRP size, (ii) had a facilitatory role in recovery from synaptic depression, and that (iii) SV2A deficits were associated with aberrant Ca2+ current density, which may contribute to the secretory phenotype in sympathetic peripheral neurons.

Scanning Electron Microscopic Preliminary Study of the Efficacy of SmearClear and EDTA for Smear Layer Removal after Root Canal Instrumentation in Permanent Teeth

This study aimed to evaluate the efficacy of SmearClear (SybronEndo, Orange, CA) and EDTA for smear layer removal from root canals of permanent teeth after instrumentation. Thirty extracted human permanent teeth (n = 10) were randomly assigned to the following groups: group 1 = 14.3% EDTA, group 2 = SmearClear, and group 3 = no smear layer removal procedure was undertaken (control). The specimens were submitted to scanning electron microscopy analysis. Magnifications of 200x and 750x were used to evaluate cleaning at the apical, middle, and cervical thirds according to a three-point scoring system. Data were analyzed statistically by the Mann-Whitney U test (5% significance level). Groups 1 and 2 differed significantly from group 3 (p < 0.01). However, there was no statistically significant difference (p > 0.05) between groups 1 and 2. In conclusion, SmearClear was able to remove the smear layer from the root canals of permanent teeth similarly as 14.3% EDTA, suggesting that both solutions may be indicated for such purpose. (J Endod 2008,34:1541-1544)

Effect of different bleaching systems on the ultrastructure of bovine dentin

The aim of this study was to evaluate in vitro the effect of different in-office bleaching systems on the surface morphology of bovine dentin. Thirty tooth fragments measuring 4 x 4mm, containing enamel and dentin, were obtained from the crowns of extracted bovine incisors. Samples were subjected to simulated intracoronal bleaching techniques using conventional (Opalescence Endo (R) and Whiteness Super Endo (R)) and light-activated systems (Opalescence Xtra (R) and Whiteness HP Maxx (R)). Controls were treated with either sodium perborate mixed with 10% hydrogen peroxide or no bleaching agent. The samples were observed under SEM and the recorded images were evaluated for topographic alterations. The ultrastructural alterations of dentin observed in this study varied greatly between groups according to the products used. Higher pH products (Whiteness HP Maxx (R) and Opalescence Xtra (R)) associated with in-office techniques yielded better maintenance of dentin ultrastructure. Apparently, both low pH and hydrogen peroxide oxidation play a role in altering the ultrastructure of dentin during internal dental bleaching. The use of alkaline products with reduced time of application (in-office techniques) may decrease such morphological alterations.