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The Journal of Neuroscience, August 11, 2004, 24(32):7174-7185; doi:10.1523/JNEUROSCI.2022-04.2004

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Cellular/Molecular
Selective Gene Expression in Magnocellular Neurons in Rat Supraoptic Nucleus

Noriko Mutsuga,¹ Tal Shahar,¹ Joseph G. Verbalis,³ Michael J. Brownstein,² Charlie C. Xiang,² Robert F. Bonner,⁴ and Harold Gainer¹

¹Laboratory of Neurochemistry, National Institute of Neurological Disorders and Stroke, National Institutes of Health, ²Laboratory of Genetics, National Institute of Mental Health, National Institutes of Health, Bethesda, Maryland 20892, ³Department of Medicine, Division of Endocrinology and Metabolism, Georgetown University, Washington, DC 20007, and ⁴Laboratory of Integrative and Medical Biophysics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892-5772

Oxytocin- and vasopressin-producing magnocellular neurons (MCNs) of the hypothalamo-neurohypophysial system are the only neuronal phenotypes present in the rat supraoptic nucleus (SON). Laser microdissection of the SON, extraction and T7-based amplification of its RNAs, and analysis of the resulting cDNAs by hybridization on a 35, 319 element DNA microarray have provided a detailed composite view of the gene expression profile of the MCNs. The genes expressed in the SON were compared with those expressed in a reference tissue consisting of total hypothalamus, and this "expression ratio" indicated which genes were preferentially expressed in the SON. Of the 26,000 unique genes on the array, 1385 were found to be expressed in the SON at levels more than two times greater than in the hypothalamus as a whole. Of these, 123 were expressed 3.4-fold higher in the SON versus hypothalamus. Most of these preferentially expressed genes were not previously known to be expressed in the MCNs. Quantitative and double-label in situ hybridization histochemistry was used selectively to confirm a number of these microarray observations and to evaluate the osmotic regulation and cell-specific expression of these genes, respectively.

Key words: oxytocin; vasopressin; hypothalamus; gene expression; microarray; laser microdissection

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Received May 25, 2004; revised June 24, 2004; accepted June 28, 2004.

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