Simona Capsoni

Reduction of regional cerebral blood flow by melatonin in young rats

Melatonin (10 ng) was subcutaneously administered to 14-day-old Sprague-Dawley rats. Regional blood flow (rCBF) was measured in 22 anatomically defined structures 20 min later using iodo[14C]antipyrine and quantitative autoradiography. rCBF was markedly reduced in the cerebral areas supplied by circle of Willis and the basilar arteries. Melatonin also significantly decreased blood flow to choroid plexuses. These findings suggest that circulating melatonin may contribute to regulation of cerebral blood flow and brain fluid balance.

Distribution and characterization of melatonin receptors in the brain of the Japanese quail, Coturnix japonica

2-[125I]iodomelatonin was used to study the distribution and properties of the melatonin receptor in the Japanese quail brain. High receptor density was detected in the major targets of direct retinal input (optic tectum, nucleus of the optic basal rout, ventrolateral geniculate nucleus), as well as areas representing terminals in the visual pathways (nucleus rotundus, ectostriatum, thalamo-hyperstriatal pathway). Binding was also found in the piriform cortex, the hypophyseal pars tuberalis, the oculomotorius nucleus and the associated Edinger-Westphal nucleus, and in the nuclei of the third, fourth and sixth cranial nerves. A comparison of the receptor pharmacological profile to that of the mammalian brain demonstrated pharmacological identity of the two binding sites. In the saturation experiments, GPT gamma S decreased the binding affinity, numerical Kd values increasing from approximately 35 pM to approximately 150 pM.

Localization and characterization of melatonin binding sites in the brain of the rabbit (Oryctolagus cuniculus) by autoradiography and in vitro ligand-receptor binding

The distribution and the properties of the melatonin binding sites were characterized in the brain of the rabbit by combined use of autoradiography and in vitro ligand-receptor binding. Autoradiography revealed widespread specific binding in the brain. The pars tuberalis of the pituitary gland, suprachiasmatic nuclei, ventromedial hypothalamic nuclei, tapetum, hippocampus, indusium griseum, cingulate gyrus, cortex and the choroid plexus were intensely labelled. Diffuse specific binding was recorded in the olfactory bulb and the anterior hypothalamus. Series of in vitro ligand-receptor binding experiments, using the anterior hypothalamus, confirmed that the binding was of high affinity and specificity. Coincubation with a non-hydrolyzable GTP analogue provoked a shift in the binding affinity, the numerical values of the Kd increasing from 20-30 pM to 280-300 pM. Apparently the melatonin receptor in the rabbit brain is linked to its second messenger via a G protein, similarly to what has been described for the brain of other vertebrates.

Characterization of endothelinA receptors in cerebral and peripheral arteries of the rat

We have characterized and quantified endothelin receptors in rat brain (anterior cerebral) and peripheral (aorta, carotid, and caudal) arteries, with the use of [125I]endothelin and quantitative autoradiography. Endothelin binding was saturable, of high affinity, and totally displaced by the selective endothelin ETA antagonist BQ 123. A single class of ETA receptors is located in the medial layer of peripheral and cerebral arteries, and its quantification by autoradiography allows study of their regulation and function.

Vasoactive intestinal peptide-like immunoreactive nerve fibers in the pineal gland of the sheep

Vasoactive intestinal peptide (VIP)‐like immunoreactive nerve fibers were demonstrated by peroxidase antiperoxidase (PAP) inmmunohistochemistry to be distributed throughout the entire pineal gland of the sheep. VIP‐containing fibers were observed along the blood vessels, penetrating into the gland from the pial capsule and also in the capsule itself. Some fibers left the perivascular position and entered the pineal parenchyma, where they were located among pinealocytes. This suggested that the VIPergic fibers might influence both pinealocytes and blood vessels of the gland. The location of VIP‐containing fibers in the capsule of the pineal gland indicates that the fibers originate from perikarya located in a peripheral ganglion.

Expression of the melatonin receptor in Xenopus laevis: A comparative study between protein and mRNA distribution

Abstract: Reverse transcription‐polymerase chain reaction (RT‐PCR) analysis performed on total RNA from different tissues of Xenopus laevis showed that the melatonin receptor gene cloned from dermal melanophores is expressed in the whole brain, skin, and retina, and that apart from the ovary, there is no expression in tissues having origin outside the central nervous system. Comparative studies using in vitro autoradiography and in situ hybridization demonstrated that the melatonin receptor is expressed with discrete allocation in Xenopus brain. Though the distribution pattern of the specific messenger RNA conforms well with that of the corresponding receptor protein, it is not always coincident.

Distribution and characterization of the melatonin receptors in the hypothalamus and pituitary gland of three domestic ungulates

Abstract: With some exceptions, in most of the mammals the pituitary pars tuber‐alis and the hypothalamic suprachiasmatic nuclei are reportedly the main targets for the pineal hormone melatonin. However, it is not known if the conspicuous diversity in the distribution pattern of melatonin binding sites in these areas depicts differences in reproductive behavior observed in the seasonally breeding species in the temperate zones. We explored the distribution and the characteristics of melatonin binding sites in the hypothalamus and pituitary of three species (bovine, horse, and donkey) different in terms of seasonal reproductive competence. The topographical localization, investigated by in vitro autoradiography, revealed 2‐[125I]iodomelatonin binding sites only in the pituitary gland in all three species, primarily in the pars tuberalis (PT), but also in the pars distalis (PD) and pars intermedia (PI). Kinetic, inhibition, and saturation studies, performed by means of in vitro binding, revealed presence of a single class high affinity binding sites. The Kd values, melatonin, and 2‐iodomelatonin Kj values were in the low picomo‐lar range. Coincubation with GTP7S inhibited 2‐[125 I]iodomelatonin binding, demonstrating that these putative receptors are linked to a G protein in their signal‐transduction pathway. The hypothalamus was devoid of specific binding. In conclusion, the results suggest that in these species, the hypophysis may be a principal target for the melatonin action on the reproductive system.

Melatonin Receptors in Brain and Peripheral Arteries

With the development of [125I]2-iodoMelatonin as a ligand, (Vakkuri et al., 1984) melatonin receptors could be localized and characterized in small, restricted brain areas (Vanecek, 1988; Laitinen et al., 1989). Melatonin receptors were soon identified by autoradiography to be present also in cerebral and caudal arteries of the rat (Viswanathan et al., 1990). In this report, we analyze the localization, characterization and regulation of these arterial melatonin receptors. Our results indicate that arterial melatonin receptors could be physiologically active, and perhaps play a role in the regulation of cerebral blood flow and body temperature.