piRNA (DQ541777) regulates dendritic spine size. (A) Rat hippocampal neurons at DIV7 were co-transfected with either LNA-based inhibitor of piRNA (DQ541777) or universal control inhibitor along with EGFP plasmid. Neurons were then imaged at DIV21 and spine size was measured based on GFP fluorescence intensity. The dendrite area chosen for high-resolution images was marked with a green box. Scale bars, 20 μm. (B) Quantitative analysis of mushroom spine area. More than 2075 spines (from 12 neurons) were analyzed in each condition to identify each spine type (mushroom, cup, stubby, and thin), and their respective area was measured. Change of spine size was observed only in mushroom spines. Mushroom spine area was calculated from more than 238 spines. The percentage change in area is represented as mean change. P < 0.05 (KS test).
