The location of the perikarya of LH-RH neurons in the rat hypothalamus and their pathways to the median eminence were studied by immunohistochemistry and radioimmunoassay after placing stereotaxic electrolytic lesions in several parts of the hypothalamus. The principal location of the cell somata was found to be in the ventral part of the medial preoptic area; their pathways were classified into a main baso-lateral pathway and an accessory descending pathway branching off from the former. The main pathway was found to cross in the vicinity of the corresponding neuronal perikarya. The central median eminence and the dorsal and ventral walls of the tubero-infundibular sulcus of the caudal part of the median eminence are innervated mainly by the baso-lateral pathway. On the other hand, the rostral and most caudal protions of the median eminence are innervated principally by the descending pathway and have a subsidiary dual innervation. The projection of LH-RH neurons to the OVLT is believed to originate from perikarya adjacent to this circumventricular organ.
Experimental procedures
Animal investigationsThe study protocol was approved by the Committee on Ethics of animal experimentation of the Japan Immunoresearch Laboratories and extra care was taken to avoid animal suffering throughout the study. Male Wistar rats (Charles River, Japan) weighing 250-300 g were fed on standard laboratory animal chow and tap water ad libitum. Before operation, the rats were anaesthetized by i,p. injection of 50 mg/kg sodium pentobarbital and then were given 1000 units/kg sodium heparin by i.v. injection.
ReagentsMCLA, a Cypridina luciferin analogue was obtained from Tokyo Kasei Co. Ltd. It was dissolved in sterile saline (pH 7.4) prior to use. The MCLA concentration was based on e430 = 9,600 M -~ .cm -~ [5]. PMA (Sigma) was dissolved in DMSO and then diluted with KrebsHensleit bicarbonate buffer (KHB), pH 7.4. GdC13 (Aldrich) was dissolved in saline (pH 7.4) just before use. Heat inactivated SOD was prepared by treating 50/,tM Cu, Zn SOD (Sigma) in an oven at 120°C for 30 rain.
FAIT exerts modest antitumor effects and poses no particular safety concerns. FAIT may be a strategy of choice worth trying for advanced HCC with high degree of vascular invasion, which is associated with poor prognosis.
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