Elena Diaz

Age differences in neurokinin A and substance P from the hypothalamus, pituitary, pineal gland, and striatum of the rat: Effect of exogenous melatonin

Previous data showed that aging of the central nervous system (CNS) is associated with widespread changes in tachykinin gene expression. However, there are no data about the possible role of exogenous melatonin in modulating the tachykinergic system during aging. The aim of this work was to analyze the age-dependent changes on neurokinin A (NKA) and substance P (SP) levels in hypothalamus, pituitary, pineal gland and striatum and the role of exogenous melatonin on these changes. We studied female rats at three different ages: 5-month-old (cyclic), 15-month-old (preacyclic) and 25-month-old (acyclic). Hypothalamic tachykinin levels increase when female rats reached acyclicity, this increase was blunted in acyclic-melatonin-treated rats. However, melatonin treatment in young cyclic rats resulted in significantly increased values as compared to controls. Pituitary NKA concentrations did no show age-dependent changes in control rats, however, in both, preacyclic and acyclic-melatonin-treated rats significantly increased values of pituitary NKA were found compared to controls. In the pineal gland, a marked decrease of NKA levels was observed in acyclic-control rats. Melatonin treatment did not alter this decrease. In the striatum, NKA and SP concentrations were significantly reduced in preacyclic- and acyclic-control rats compared to young cyclic rats, melatonin had no effect on striatal tachykinins. Our results indicate that melatonin may regulate tachykinin stores during aging mainly on structures of the neuroendocrine-reproductive axis.

Expression and regulation of Icer mRNA in the Syrian hamster pineal gland

Inducible-cAMP early repressor (ICER) is a potent inhibitor of CRE (cAMP-related element)-driven gene transcription. In the rat pineal gland, it has been proposed to be part of the mechanisms involved in the shutting down of the transcription of the gene coding for arylalkylamine N-acetyltransferase (AA-NAT, the melatonin rhythm-generating enzyme). In this study, we report that ICER is expressed in the pineal gland of the photoperiodic rodent Syrian hamster although with some difference compared to the rat. In the Syrian hamster pineal, Icer mRNA levels, low at daytime, displayed a 20-fold increase during the night. Nighttime administration of a beta-adrenergic antagonist, propranolol, significantly reduced Icer mRNA levels although daytime administration of a beta-adrenergic agonist, isoproterenol, was unable to raise the low amount of Icer mRNA. These observations indicate that Icer mRNA expression is induced by the clock-driven norepinephrine release and further suggest that this stimulation is restricted to nighttime, as already observed for Aa-nat gene transcription. Furthermore, we found that the daily profile of Icer mRNA displayed photoperiodic variation with a lengthening of the nocturnal peak in short versus long photoperiod. These data indicate that ICER may be involved in both daily and seasonal regulation of melatonin synthesis in the Syrian hamster.

Effect of prenatal melatonin on the gonadotropin and prolactin response to the feedback effect of testosterone in male offspring

The purpose of this study was to investigate the effects of prenatal melatonin administration on the sensitivity of the androgens negative feedback effect on gonadotropin and prolactin secretion in male offspring. Male offspring of control (control-offspring) and melatonin treated (MEL-treated) (150 microg/100 g BW) mother rats during pregnancy (MEL-offspring), at infantile, prepubertal, and pubertal periods were studied. LH secretion in response to testosterone propionate (TP) in control-offspring showed the classical negative feedback effect at all ages studied. In MEL-offspring a negative response after TP was also observed in all ages studied although the magnitude of this response was altered in this group as compared to controls. FSH values were significantly lower at most ages and time points studied in MEL-offspring than in control-offspring. FSH secretion in MEL-offspring showed a delayed negative feedback action of TP injection as compared to control-offspring. This response was observed at 21 days of age in control-offspring and delayed until day 30 of life in MEL-offspring. Parallely it remain at later age in MEL-offspring than in control-offspring. Prolactin secretion in control-offspring showed increased values after TP injections from infantile to pubertal periods. This increase was blunted in MEL-offspring at 17 and 35 days of age showing significantly reduced (p<0.01; p<0.05) plasma prolactin levels. During pubertal period a prolactin positive response to TP administration was observed in MEL-offspring but with significantly lower magnitude than in control-offspring. These results indicate that prenatal melatonin exposure induced changes in the sensitivity of gonadotropin and prolactin feedback response to testosterone, indicating a delayed sexual maturation of the neuroendocrine-reproductive axis in male offspring.

Evidence for clock genes circadian rhythms in human full-term placenta

Abstract Biological rhythms are driven by endogenous biological clocks; in mammals, the master clock is located in the suprachiasmatic nucleus (SCN) of the hypothalamus. This master pacemaker can synchronize other peripheral oscillators in several tissues such as some involved in endocrine or reproductive functions. The presence of an endogenous placental clock has received little attention. In fact, there are no studies in human full-term placentas. To test the existence of an endogenous pacemaker in this tissue we have studied the expression of circadian locomoter output cycles kaput (Clock), brain and muscle arnt-like (Bmal)1, period (Per)2, and cryptochrome (Cry)1 mRNAs at 00, 04, 08, 12, 16, and 20 hours by qPCR. The four clock genes studied are expressed in full-term human placenta. The results obtained allow us to suggest that a peripheral oscillator exists in human placenta. Data were analyzed using Fourier series where only the Clock and Bmal1 expression shows a circadian rhythm.

Prenatal melatonin exposure affects luteinizing hormone and hypothalamic and striatal neuropeptide Y in the male rat offspring

The present study examines the influence of prenatal melatonin on the hypothalamic and striatal neuropeptide Y (NPY) concentrations as well as on luteinizing hormone (LH) levels. Male rat offspring of control and melatonin treated mother rats were studied at different ages of the sexual development: infantile, prepubertal, pubertal and adult ages. Hypothalamic NPY levels were much higher during the juvenile than throughout the infantile period. After prenatal melatonin treatment significantly higher values since day 15 up to 35, also at 60 days of age were found, as compared with controls. Striatal NPY levels were lower than in hypothalamus. Again, NPY in the striatum from offspring of melatonin treated mother rats showed significantly higher values than the respective controls at most of the ages studied. However, prenatal melatonin exerted an inhibitory influence upon LH secretion pattern, since decreased concentrations up to 25 days of age and delayed peak values at pubertal age were observed. The present study also suggest that the effect of NPY upon LH secretion is related to sexual development, since NPY exerted opposite effect in infantile than in pubertal period and melatonin administration during intrauterine life prevented this effect.

Prenatal melatonin exposure influences the maturation of gonadotropin and prolactin estradiol-benzoate feedback system

Gonadotropin and prolactin response to estrogen feedback in female rat offspring of control and melatonin treated (150 microg/100 g BW) mother rats during pregnancy (MEL-offspring) were studied at these periods: infantile, prepubertal and pubertal. In controls negative or absent LH feedback developed after estradiol benzoate (EB) injection up to 30 days of age indicating that the onset of puberty had not occurred. The positive feedback was established from day 33 on. However, in MEL-offspring the first activation of gonadotropin secretion during afternoon, 31 h after EB, was observed at 25 days of age, representing the first neuroendocrine sign of the onset of puberty. This positive response disappeared on day 30 in MEL-offspring. At 33 days of age, the LH positive response to EB was found in both groups, indicating a more advanced sexual development. In controls, this response increased at 35 days of age while in MEL-offspring it was highly depressed. FSH secretion in response to EB showed a negative feedback effect from infantile to the end of prepubertal period in both groups. The positive feedback was observed earlier in MEL-offspring (at 33 days of age) than in controls (at 35 days of age), but at this age it was absent in MEL-offspring. A positive prolactin response to EB at all ages in controls was observed. The typical pulsatility with higher values in the afternoon appeared by the first time at 30 days of age. However, in MEL-offspring no pulsatile response was observed throughout any age. These data suggest that prenatal melatonin administration altered gonadotropin and prolactin response to EB inducing precocious sensitivity during prepubertal period but depressed response during the pubertal period.

Circadian urinary citrate excretion in a rat model of exercise

Aims: Circadian rhythms are the approximate 24 h biological cycles that function to prepare an organism for daily environmental changes. Circadian rhythms unquestionably play critical roles in metabolic homeostasis and the exercise has emerged as a strong non‐photic time cue or zeitgeber in animal models and humans. Numerous studies about the effects of exercise on the citrate synthase activity have been published. Citrate is used to assess energy production or expenditure because it is a substrate of the Krebs Cycle, a cycle for oxidative energy production. Main methods: We tested the existence of a rhythmic urinary citrate excretion in a rat model that is made to exercise at six different points during the day. Key findings: The data obtained by the enzyme assays were fitted to a mathematical model (Fourier series), showing for the first time, the existence of a distinct ultradian rhythm in the urinary citrate excretion. The aerobic exercise led to the increase in the period length of the ultradian rhythm and raised the acrophase value of the urinary citrate excretion. Therefore, the urinary citrate excretion pattern changed after exercise, showing a clear circadian rhythm fitted to the mathematical model. Significance: The citrate urine samples could provide accurate data for ranking an individuals metabolic status. Using exercise to maintain the circadian clock at an appropriate phase and amplitude might be effective to prevent cardiometabolic disease development.

Circadian Kisspeptin expression in human term placenta

Kisspeptin is an essential gatekeeper of reproductive function. During pregnancy high circulating levels of kisspeptin have been described, however the clear role of this neuropeptide in pregnancy remains unknown. We tested the existence of rhythmic kisspeptin expression in human full-term placenta from healthy pregnant women at six different time points during the day. The data obtained by Western blotting were fitted to a mathematical model (Fourier series), demonstrating, for the first time, the existence of a circadian rhythm in placental kisspeptin expression.

Ovarian Aging: Melatonin Regulation of the Cytometric and Endocrine Evolutive Pattern

We have investigated the effect of melatonin administration on the cytometric and endocrine functions of the ovary during aging. Young cyclic (3 months old), middle-aged pre-acyclic (13 months old), and old acyclic (22 months old) female Wistar rats were used for two months, in both control and melatonin-treated groups. Cell cycle by flow cytometry: the percentage of ovarian cells in the G0-G1 phase was the highest in both control and melatonin-treated rats. However, melatonin treatment significantly reduced (P< 0.05) the percentage of cells in the G0-G1 phase compared to control rats. This reduction of cells in the G0-G1 phase is derived to the S phase in cyclic and acyclic rats. The possibility that a tumoral process leads to a proliferative effect observed in young and acyclic melatonin-treated rats was ruled out because no significant differences were found for p53 and Ki67 expression levels between control and melatonin groups. Density of oocytes: the oocyte number per ovary unit volume was not affected by melatonin treatment in the three age ranges studied. Melatonin treatment in middle-aged (pre-acyclic) rats resulted in significantly higher (P< 0.05) ovarian volume; higher oocyte volume, without significant differences, and oocytes in circular form were significantly (P< 0.05) higher than in control rats. Melatonin treatment during pre-acyclic age range could resynchronize the estrous cycle periodicity. Melatonin treatment was able to maintain the same levels of estradiol in the pre-acyclic age groups studied as those observed in the young cyclic rats. The present results indicate that melatonin administration to middle-aged female rats produces beneficial effects that extend the reproductive function of the ovary.

Efecto de la glándula pineal materna en las variaciones hormonales estacionales del eje neuroendocrino-reproductor de los descendientes

Background and objective To investigate the influence of the maternal pineal gland on seasonal variations in luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin, as well as on body weight in offspring. Subjects and method Female Wistar rats maintained in 12 h light/12 h dark were used. Mother rats were divided in the four seasons of the year into three groups: control, pinealectomized (PIN-X) and PIN-X plus melatonin treatment (PIN-X+MEL). Male offspring at 21, 31 and 60 days of age were studied in the four seasons of the year. Results Maternal PIN-X resulted in increased body weight at 21 and 31 days of age in winter and at 60 days of age in spring and summer. Maternal PIN-X altered LH developmental pattern in the prepubertal period in fall and winter but the effect of PIN-X + MEL treatment was observed only in spring and summer. Maternal PIN-X significantly Lowered FSH values at 60 days of age in summer and fall. The developmental pattern of prolactin was not affected by maternal PIN-X but a slight influence of PIN-X + MEL treatment on both hormones was observed in a seasonal dependent manner. Conclusions Our results suggest that during fetal development maternal melatonin may affect postnatal development of the neuroendocrinereproductive axis in a seasonal dependent manner. These considerations may help to elucidate questions related to seasonal disorders, which are becoming more and more frequent.