Abstract
Publisher Summary Histological localization of RNA involves in situ hybridization techniques. Light microscopy (LM) in situ hybridization techniques have allowed localization of messenger RNAs (mRNAs) to specific regions of the cell. The mechanism of intracellular RNA localization involves a two-step process: transport from the perinuclear region and then anchoring of the RNA to a specific subcellular location. Transport processes involve assembly of the RNA with cellular factors and movement of the complex along microfilaments and/or microtubules. In contrast little is known about the anchoring process. To identify the cellular structures that provide RNA anchoring sites, high resolution in situ hybridization techniques at the electron microscopy (EM) level are required. The success of EM in situ hybridization relies on experimental conditions for production of high specific activity nonisotopic probes, saturated hybridization kinetics, and efficient detection of the hybridization signal. This chapter describes a protocol for detecting the nonrandom distribution of rice storage protein mRNAs on the endoplasmic reticulum (ER) using nonradioactive RNA probes.
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