[Recent advances in molecular histochemical techniques]

Abstract

To understand better the physiological states of cells and tissues, it is essential to analyze the regulatory mechanism of cell-type or tissue-type specific gene expression in individual cells. For this purpose, in situ hybridization (ISH) has been regarded as a powerful molecular histochemical method. Over the past decade, we made lots of efforts to establish nonradioactive ISH protocol with thymine-thymine dimerized or digoxigenin-labeled synthetic oligo-DNA probes, including extensive optimization of pretreatment, evaluation of hybridizable RNA and new trials of signal amplification. Furthermore, we have originally developed a new method, Southwestern histochemistry, which localizes specific transcription regulatory factors and allows us to understand the fluctuation of mRNA levels detected by ISH at the level of transcription regulation of the gene in individual cells. Here, I will review the outlines of these methods with recent progress and will see what we can see by combining them.