Rabbit Kidney Cells with mEmerald-α-Actinin and mApple-Lysosomes
Lysosomes vary in size and shape, but are all are contained by a single layer membrane. This membrane shields the remaining cell from the lysosomes' harsh digestive enzymes. Lysosomes provide additional support to the cell with its pH of about 4.8, which means the neutral environment of the cytosol makes most of the digestive enzymes inoperative, so even if a lysosome bursts, the holistic cell should remain uninjured. The lysosome's acidity is maintained with the help of hydrogen ion pumps. The proximity between lysosomes and the F-actin cytoskeletal network is examined in this digital video sequence.
The featured rabbit kidney epithelial cells (RK-13 line) are shown expressing mApple fused to lysosomes and mEmerald fused to alpha-actinin, an actin-bundling protein. mApple is a red member of the mFruit series of fluorescent proteins selected for enhanced photostability. Peak excitation and emission of mApple occur at 568 and 592 nanometers, respectively. The green fluorescent protein mEmerald is one of many variants of the enhanced Aequorea derivative EGFP. mEmerald has a peak excitation wavelength of 487 nanometers and peak emission wavelength of 509 nanometers.