Published online by Cambridge University Press: 18 September 2015
In vitro biosynthesis of thyrotropin releasing factor (TRF) by different regions of the hypothalamus of mink was examined. Homogenates of hypothalamic tissue were incubated in Krebs-Ringer medium containing 200 mg% glucose, 10-5M ATP, 0.1 mM histidine and glutamic acid and 0.15 μ c 3H-proline (40 Cilmmol) per mg. tissue. Extraction, purification and estimation of 3H-TRF biosynthesis involved several steps of charcoal extraction, carboxymethylcellulose and sephadex chromatography. 3H-TRF was synthesized throughout the entire antero-posterior extent of the hypothalamus in its dorsal and medial portions. 3H-TRF was synthesized also in a more discreet region, the arcuate nucleus. In vitro biosynthesis of 3H-TRF was stimulated significantly by thyroxine, but not by TSH, estradiol, corticosterone or melatonin. A method is described for collection of cerebrospinal fluid of the third ventricle of the rat brain; TRF concentration in this fluid was approximately in normal animals.
The distribution of TRF-producing cells in the hypothalamus and presence of TRF in cerebrospinal fluid of the third ventricle is discussed with respect to the hypothesis that this releasing factor may be delivered to the median eminence and adenohypophysis in part, via the cerebrospinal fluid.