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The circumventricular organs of the frog Rana c(n‘esbefan(1 were studied at the light and electron microscopic levels with antisera against catecholamine-synthesizing enzymes (tyrosine hydroxylase: TH, dopamine-fi-hydroxylase: DBH, phenylethanolamine-N-methyltransferase: PNMT) and serotonin (5-HT). The present study is the first to demonstrate the presence of three groups of neuronal vesicles in the liquorcontacting neurons which are either TH or 5-HT-positive. The neurons contacting the preoptic recess liquor were TH-positive but DBH-negative. Amounts of THimmunoreactive product were localized not only in the cytoplasm but also in the granules (90-100, 120-150, and 170-200 nm in diameter). This suggests that dopamine (DA) synthesis occurs in the cytoplasm as well as in cored vesicles. A few 5HT-positive, TH-negative serotonergic neurons were scattered in the preoptic recess organ. In the paraventricular organ, subependymal liquor-contacting neurons were shown to be serotonergic. In immunoelectron microscopy, 5-HT-immunoreactivity was localized both in the cytoplasm and in the cored vesicles (90~l20, 130-170, and 200-210 nm in diameter). I11 the rneclullospinal liquor-contacting neurons, the fourth ventricular organ, TH-positive and DBH-positive, but PNMT-negative and 5-I-IT-negative reactions were observed in the granules (70-80, 90-100, and 110-130 nm in diameter). Noradrenaline (NA) seemed also to be present in the cored vesicles. It is concluded from the above results that cored vesicles may contain DA and 5-HT besides NA. Studies on the aminergic system of the amphibian brain to date (1, 2, 7, 15, 16, 21, 23, 25) have been performed by the histofluorescence method. Since the studies of Vigh er al. (23), VighTeichmann er 01. (25) and Peute (18, 19), the circumventricular organs of the frog have been examined by light and electron microscopy (2, 7, 15, 24). The histofluorescent cells were believed to contain monoamines, which Braak (1) identified as catecholamines and serotonin (5HT). Braak ( 1) reported that both monoamines were localized in the nucleus reticularis mesencephali, but that only catecholamines were localized in the organum vasculosum hypothalami and in the area praeoptica. Later, Parent (16) and other researchers described catecholamine neurons in the frog paraventricular organ and preoptic recess organ. On the other hand, Prasada Rao and Hartwig (21) found both catecholamine and 5-HT in frog paraventricular organ by microspectrofluorirnetry. In formaldehyde-induced histofluorescent studies, differentiating between dopamine (DA) E

biomedical research

&lt;b&gt;IMMUNOHISTOCYTOCHEMICAL DISTRIBUTION OF AMINE-CONTAINING NEURONS RELATED TO CIRCUMVENTRICULAR ORGANS IN THE FROG &lt;/b&gt;&lt;b&gt;BRAIN &lt;/b&gt;