Quantitativein situ hybridization to measure single-cell changes in vasopressin and oxytocin mRNA levels after osmotic stimulation
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The measurement of cellular mRNA content by quantitativein situ hybridization is a valuable approach to the study of gene expression in brain since this tissue exhibits a high degree of phenotypic heterogeneity.
The cellular content of vasopressin and oxytocin mRNA in hypothalamo-neurohypophysial system neurons was altered by maintaining rats for 24 hr on 2% sodium chloride water.
Statistical and graphical techniques were then used to analyze cell by cell how mRNA levels were altered as a result of osmotic stimulation. We propose that the negative binomial probability distribution is a suitable model to describe how mRNA content varies across a defined cell population. For both measures of oxytocin and vasopressin mRNA levels, maximum-likelihood estimation indicated that this model adequately described empirical findings obtained from rats drinking tap water or salt water.
Both graphical and statistical analyses suggested how the defined neural system responds to osmotic stimulation: mRNA content was altered as a multiplicative function of “initial state.” The utility and limitations of the quantitative approach are discussed.
Key wordsautoradiography morphometry biological models in situ hybridization neurohypophysial system supraoptic nucleus vasopressin oxytocin messenger RNA
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