Imaging a specific mRNA in pollen with atomic force microscopy

Abstract

Intracellular localization of RNA is important in regulating various cellular functions. Here, we determined the intracellular localization of the ammonium transporter (AtAMT1;4) mRNA in the sperm cells of Arabidopsis pollen by atomic force microscopy (AFM). The AFM probe was modified with cone-shaped dendron molecules, and an oligonucleotide complementary to the mRNA was attached to the apex. This configuration allowed facile detection of the mRNA within a sectioned pollen cell at the single-molecule level by measuring the mRNA–oligonucleotide interaction force. The AtAMT1;4 mRNA was largely localized in the nucleus rather than the cytosol of the sperm cells.