Julieta Helena Scialfa

The effects of lesions of the thalamic intergeniculate leaflet on the pineal metabolism

The aim of the present work was to study, in rats, the effects of lesions of the thalamic intergeniculate leaflet (IGL) and the deep pineal/lamina intercalaris region (DP) on the diurnal profile of N-acetylserotonin (NAS) and on the nocturnal pineal reactivity to acute retinal light stimulation (1 or 15 min). The 24-h experiment shows that there is no phase-shifting on the diurnal NAS curve of groups of rats with bilateral IGL lesion compared to the controls. On the other hand there is a significant reduction on the amplitude of pineal NAS content observed in every nocturnal point of the curve. The pineal glands of IGL-lesioned rats, after 1 min of retinal light stimulation, keep their NAS content equal to the lesioned dark-killed rats. Nonetheless, after 15 min of photostimulation, the pineal NAS content is reduced to nearly zero equally to the control animals. DP lesion does not modify the content of NAS in the pineal gland of rats killed in the dark. However, the pineal photo-inhibition process induced by 1 min of light exposure is impaired. These results suggest that: (1) the intergeniculate leaflet has a role in regulating the amplitude of the diurnal rhythm of pineal NAS production rather than its phase entrainment to light-dark cycle. This effect is not dependent on the direct geniculo-pineal connections. (2) The nocturnal pineal photo-inhibition phenomenon could be decomposed in two processes. One, triggered by short pulses of light and totally dependent on the IGL and partially dependent on the direct monosynaptic pathway between this structure and the pineal gland.(ABSTRACT TRUNCATED AT 250 WORDS)

Melatonin synthesis impairment as a new deleterious outcome of diabetes‐derived hyperglycemia

Melatonin is a neurohormone that works as a nighttime signal for circadian integrity and health maintenance. It is crucial for energy metabolism regulation, and the diabetes effects on its synthesis are unresolved. Using diverse techniques that included pineal microdialysis and ultrahigh‐performance liquid chromatography, the present data show a clear acute and sustained melatonin synthesis reduction in diabetic rats as a result of pineal metabolism impairment that is unrelated to cell death. Hyperglycemia is the main cause of several diabetic complications, and its consequences in terms of melatonin production were assessed. Here, we show that local high glucose (HG) concentration is acutely detrimental to pineal melatonin synthesis in rats both in vivo and in vitro. The clinically depressive action of high blood glucose concentration in melatonin levels was also observed in type 1 diabetes patients who presented a negative correlation between hyperglycemia and 6‐sulfatoxymelatonin excretion. Additionally, high‐mean‐glycemia type 1 diabetes patients presented lower 6‐sulfatoxymelatonin levels when compared to control subjects. Although further studies are needed to fully clarify the mechanisms, the present results provide evidence that high circulating glucose levels interfere with pineal melatonin production. Given the essential role played by melatonin as a powerful antioxidant and in the control of energy homeostasis, sleep and biological rhythms and knowing that optimal glycemic control is usually an issue for patients with diabetes, melatonin supplementation may be considered as an additional tool to the current treatment.

Tryptophan hydroxylase is modulated by L-type calcium channels in the rat pineal gland

Calcium is an important second messenger in the rat pineal gland, as well as cAMP. They both contribute to melatonin synthesis mediated by the three main enzymes of the melatonin synthesis pathway: tryptophan hydroxylase, arylalkylamine N-acetyltransferase and hydroxyindole-O-methyltransferase. The cytosolic calcium is elevated in pinealocytes following alpha(1)-adrenergic stimulation, through IP(3)-and membrane calcium channels activation. Nifedipine, an L-type calcium channel blocker, reduces melatonin synthesis in rat pineal glands in vitro. With the purpose of investigating the mechanisms involved in melatonin synthesis regulation by the L-type calcium channel, we studied the effects of nifedipine on noradrenergic stimulated cultured rat pineal glands. Tryptophan hydroxylase, arylalkylamine N-acetyltransferase and hydroxyindole-O-methyltransferase activities were quantified by radiometric assays and 5-hydroxytryptophan, serotonin, N-acetylserotonin and melatonin contents were quantified by HPLC with electrochemical detection. The data showed that calcium influx blockaded by nifedipine caused a decrease in tryptophan hydroxylase activity, but did not change either arylalkylamine N-acetyltransferase or hydroxyindole-O-methyltransferase activities. Moreover, there was a reduction of 5-hydroxytryptophan, serotonin, N-acetylserotonin and melatonin intracellular content, as well as a reduction of serotonin and melatonin secretion. Thus, it seems that the calcium influx through L-type high voltage-activated calcium channels is essential for the full activation of tryptophan hydroxylase leading to melatonin synthesis in the pineal gland.

Pineal metabolic reaction to retinal photostimulation in ganglionectomized rats

The aim of the present work was to test the pineal gland metabolic reactivity to nocturnal retinal short term photic stimulation in superior cervical ganglionectomized rats. The experimental support for this work is the appearance of a transitory post synaptic hyperactivity in the pineal gland, during the anterograde degenerating process of the conarii sympathetic nerve fibers after surgical removal of the cell body. In this situation the pineal gland is deafferented from the peripheral sympathetic nervous system keeping intact, however, the direct central connections to the deep pineal/lamina intercalaris region (DP). The results show a blockade of the pineal noradrenergic stimulatory process due to the retinal photostimulation. The inactivation of N-acetyltransferase led to a true metabolic shift to the oxidative pathway resulting in a decrease of the amount of N-acetylserotonin and an increase of the amount of serotonin, 5-hydroxyindoleacetic acid and 5-hydroxytryptophan. This inhibitory process brought into action by retinal illumination is dependent on the direct central neural connections to the pineal gland, since rats that were lesioned in the DP, previously to ganglionectomy, did not show any alteration on the indolic content of the pineal gland when subjected to nocturnal retinal photostimulation.

Lesions of the Dorsomedial Hypothalamic Nucleus Do Not Influence the Daily Profile of Pineal Metabolism in Rats

The present study attempted to characterize the effects of electrolytic lesions of the hypothalamic dorsomedial nucleus on the daily profile of pineal metabolism as well as on the inhibition of pineal melatonin synthesis induced by acute light exposure during the night. Adult male Wistar rats (n = 107, 12:12 h light-dark cycle) were left intact (n = 47) or lesioned (n = 60). Lesioned rats and their respective controls were killed at six time points distributed throughout the light-dark cycle. At ZT (zeitgeber time) 18 the animals were killed either in the dark or after 15 min of light stimulation. Pineal glands were assayed using high-performance liquid chromatography with electrochemical detection (HPLC-ED). There was no difference in the amounts of pineal indoles between lesioned and control rats under any of the experimental situations tested. These results suggest that in rats, the hypothalamic dorsomedial nucleus does not participate in either the neural control of daily pineal metabolism or the nocturnal light-induced inhibition of the pineal metabolism.

Enhancement of sodium intestinal secretion in relation to absorption in malnourished rats: hyperosmolar challenge

Two experimental models were tried in young malnourished rats in order to study effect of an hyperosmolar challenge in the small intestine on the bi-directional fluxes of sodium. Weanling rats were fed with energy restricted diets. In model I 1 mL of NaCl 900 mOsm/kg was introduced in the small intestine of the rats and left from 5 up to 70 min, in order to determine the moment of higher net Na+ secretion, which occurred at 10 min. In model II, the bi-directional fluxes of Na+ and Cl- were studied using NaCl or mannitol 900 mOsm/kg under the effect of mecholil, atropine or 2-4 dinitrophenol, for 10 min. Mecholil decreased the Na+ absorption enhancing the net secretion. Control rats were used as reference. In the restricted diets animals occurred an increase of the net secretion stimulated by NaCl 900 mOsm/kg, and this effect was enhanced by mecholil. It is suggested that in malnutrition there is an impairment in Na- intestinal absorption.

Dietary nitrite and scavenger antioxidants trace elements

Rats initially weighing 138 +/- 14 g were fed the following diets for 150 days: control (Co), control plus nitrite-bacon-proline 24 mg/kg, 100 g/kg and 10 mg/kg, respectively (NB), NB plus 0.04 micrograms/g selenium (NBSe) and NB plus 0.020 g/kg ascorbic acid (NBC). The NB diet provoked body weight and feeding efficiency enhancement with a reduction in body density increasing serum lactic acid, uric acid and cholesterol levels. The serum selenium decreased by the presence of NB in the diet. The addition of selenium and ascorbic acid to the NB diet prevented the reduction in body density and also affected uric acid and cholesterol levels. It is suggested that the NB diet has adverse effects and that some of the alterations it causes are prevented by the reducing elements selenium and ascorbic acid.

Selenium as protector of degenerative diseases

Antioxidants are scavengers of free radicals related to atherosclerosis and cancer, and probably selenium as a component of glutathione peroxidase (GSH-Px) plays an important role in this mechanism. Young rats of average initial body wt of 100 g were fed for 60 d with the following diets: I-Casein, complete, selenium free (0.0003 I~g/kg), 17% of protein (Cas) or with Se added (0.004 ~g/kg) (Cas.Se); II-As I, but with nitrite (24 mg/kg) and bacon (100 g/kg) added (Cas-NB), selenium free (0.0003 i~g/kg) or with Se added (Cas-NB-Se). The main results showed that: