Development of an organ culture technique capable of monitoring most pineal gland indole metabolites.

An intact pineal gland organ culture technique was developed which utilised radiolabelled tryptophan as the indolic precursor and two-dimensional thin-layer chromatography to separate the various indole metabolites produced. The method was capable of reproducibly separating and quantitating all tryptophan metabolites except 5-methoxytryptophan which co-chromatographed with tryptophan in all the solvent systems evaluated. Noradrenergic stimulation of cultured pineals led to a predictable increase in N-acetyl-serotonin and melatonin production, suggesting that the method was useful for biochemical and pharmacological studies on the pineal gland. Similarly evaluation of the results revealed that a strong linearity existed between N-acetylserotonin and melatonin production and between actual and theoretical methylation as previously reported, again verifying the usefulness of the method developed.