In situ hybridization analysis of osmotic stimulus-induced changes in ribosomal RNA in rat supraoptic nucleus.

A quantitative in situ hybridization analysis was used to investigate changes in levels of ribosomal RNA (rRNA) in neurons of the supraoptic nucleus (SON) of rats stimulated osmotically by giving 2% NaCl as drinking solution for 0 (control rats), 1, 4, and 14 days. The quantitation was autoradiographically accomplished by in situ hybridization with a nick-translated tritiated ribosomal DNA probe and with the use of computer-based image analysis system. The mean number of grains per neuron in the ventral SON was significantly increased: 1.8-fold for 1 day, 2.9-fold for 4 days, and 1.7-fold for 14 days of salt loading, whereas the mean number of grains per neuron in the dorsal SON was increased 1.3-fold for 1 day, 2.5-fold for 4 days, and 1.7-fold for 14 days. Kolmogorov-Smirnov analysis of frequency histograms of grains per neuron indicated that the amount of rRNA in neurons in the ventral and dorsal SON was significantly increased by osmotic stimulation. These increases were accompanied by increases in cell size. The subcellular location of hybridizable rRNA in magnocellular neurons was altered by osmotic stimulation. Following 1-14 days of salt-drinking, rRNAs appeared to be more unevenly distributed throughout the cytoplasm. These findings are consistent with the notion that hyperosmotic stimulation has a substantial effect on the expression of rRNA genes in neurons of both the ventral and dorsal SON.