Skip to main content
. 2008 Aug 11;105(33):11720–11724. doi: 10.1073/pnas.0804015105

Fig. 3.

Fig. 3.

Sialin was associated with pineal SLMVs and is involved in exocytosis of aspartate and glutamate. (A) Sialin was present in pineal SLMV fractions as revealed by immunoblotting. The position of sialin is marked by an arrow. (B) Sialin is colocalized with aspartate and synaptophysin. Cultured pinealocytes were double immunostained with antibodies against aspartate and synaptophysin (Upper) or sialin and synaptophysin (Lower). (Scale bar, 10 μm.) (C) Double-labeling immunoelectron microscopy demonstrated the association of sialin with synaptophysin. Samples were treated with sets of anti-synaptophysin monoclonal antibodies (5-nm particles, arrows) and anti-sialin serum (10-nm particles, arrowheads) or anti-synaptophysin monoclonal antibodies (5-nm particles, arrows) and anti-VGLUT2 serum (10-nm particles, arrowheads) or anti-synaptophysin monoclonal antibodies (5-nm particles) and control serum (10-nm particles). (Scale bar, 100 nm.) (D) RNAi directed against rat sialin decreased sialin expression without affecting the expression of VGLUT2. Quantitative analysis for sialin and VGLUT2 mRNA levels was performed by real-time PCR. (E) Exocytosis of aspartate and glutamate after 20-min incubation upon KCl stimulation from control siRNA-treated pinealocytes and sialin siRNA-treated pinealocytes.