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Komajda, and K. Schwartz, J. Clin. Invest. 85, 305 (1990). 14. D. de la Bastie, D. Levitsky, L. Rappaport, J. J. Mercadier, F. Marotte, C. Wisnewsky, V. Brovkovich, K. Schwartz, and A. M. Lompre, Circ. Res. 66, 554 (1990). [3] Molecular Assays for Rat ThyrotropinReleasing Hormone Gene Stephanie L. Lee and Kevin A. Sevarino Introduction Thyrotropin-releasing hormone (TRH) is central to the control of the hypothalamic-pituitary-thyroid axis. TRH is present throughout the external zone of the median eminence in axon terminals that originate from a group of neurons located in the paraventricular nucleus (PVN), a region that corre­ sponds to the "thyrotropic area" of the hypothalamus.

The bag is sealed by replacing the cap on the tubing and placed in a box containing water preequilibrated at 42°C in an agitating water bath. Prehybridization is performed at 42°C for at least 4 hr with slow agitation. After prehybridization, the buffer is removed from the bag with a syringe and replaced with an equal volume of fresh buffer to which the probe has been added. Since the probe is single-stranded, there is no need to heat it prior to addition to the buffer. The bag is placed back in the box and agitated overnight at 55°C in the water bath.

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