Rabbit Kidney Cells with mEmerald-Actin and mApple-Lysosomes
There are multiple vital enzymes found in lysosomes, including lipase, which digests lipids; amylase, which digest carbohydrates; proteases, which digest proteins; nucleases, which digest nucleic acids; and phosphoric acid monoesters. Lysosomal enzymes are synthesized in the endoplasmic reticulum and the cytosol where they are targeted for the lysosome by a mannose-6-phosphate tag. Aberrant lysosomal targeting leads to inclusion-cell disease, or when enzymes do not properly reach the lysosome. This results in waste accumulation within these organelles.
Lysosomes were fluorescently labeled with the red fluorescent protein mApple in the rabbit kidney epithelial cells (RK-13 line) appearing in this digital video and actin was visualized with the green fluorescent protein mEmerald. mApple, like other members of the mFruit series of fluorescent proteins, was derived from mRFP1 using a directed evolution approach. Excitation of mApple peaks at 568 nanometers and emission peaks at 592 nanometers. mEmerald is an enhanced variant of EGFP that exhibits improved brightness and photostability. Excitation and emission maxima of mEmerald occur at 487 and 509 nanometers, respectively.