In the pineal of 21-day old male Wistar rats hydroxyindole-O-methyltransferase (HIOMT) activities involved in the synthesis of several 5-methoxyindoles were determined during the night in April, June, October and January. A high HIOMT activity for the synthesis of melatonin/5-methoxytryptophol was determined in the months of January and April. In June and October a decrease was observed. The activity maxima coincide with peaks of activity found for the synthesis of 5-methoxytryptophan. Synthesis of 5-methoxytryptamine occurred only in June and October, whereas the synthesis of 5-methoxyindole-3-acetic acid occurred only in January. From these results it may be concluded that January and April are the most active months of those tested for the melatonin/5-methoxytryptophol synthesis in the rat pineal gland. A possible physiological role of the 5-methoxyindoles other than melatonin and 5-methoxytryptophol is discussed.

Summary. Using a method in which no substrate is added, the capacity of HIOMT to synthesize melatonin and/or 5-methoxytryptophol has been determined in the pineal and the eyes of the mole, a mammal having an atrophied visual system. In all animals studied, the activity of HIOMT in both eyes taken together was 2-10 times higher than in the pineal. The results prove that the pineal is not the only, and not always the most important, source of methoxyindoles.Introduction.

In the present paper, an extension is presented of an earlier described method, by which the methylating capacity of the pineal gland can be determined. Supplementary to the earlier method, the synthesis of melatonin and 5-methoxytryptophol can now be qualified and quantified separately.

In pineals of 10 day old rats 5-hydroxytryptophan, 5-hydroxyindole-3-acetic acid, N-acetylserotonin/5-hydroxytryptophol and norepinephrine are methylated following a circadian rhythm. During the night HIOMT and COMT activities were measured for the above mentioned substrates, while HIOMT activity for 5-hydroxytryptophan and N-acetylserotonin/5-hydroxytryptophol was also determined during daytime.

In order to obtain more information on the methylating capacity of the pineal gland, a method determining the formation of different 5-methoxyindoles in the pineal gland was developed. The method depends on measuring the incorporation of labelled methyl groups into the various hydroxyindoles present in the pineal gland, after incorporation of pineal tissue with labelled S-adenosyl methionine. Hydroxyindoles were not added to the incubation medium. After incubation thin-layer chromatography was performed with pineal tissue together with the incubation medium; the spots were scraped and counted.