Fernando Torrealba

Antiribosomal-P autoantibodies from psychiatric lupus target a novel neuronal surface protein causing calcium influx and apoptosis

The interesting observation was made 20 years ago that psychotic manifestations in patients with systemic lupus erythematosus are associated with the production of antiribosomal-P protein (anti-P) autoantibodies. Since then, the pathogenic role of anti-P antibodies has attracted considerable attention, giving rise to long-term controversies as evidence has either contradicted or confirmed their clinical association with lupus psychosis. Furthermore, a plausible mechanism supporting an anti-P–mediated neuronal dysfunction is still lacking. We show that anti-P antibodies recognize a new integral membrane protein of the neuronal cell surface. In the brain, this neuronal surface P antigen (NSPA) is preferentially distributed in areas involved in memory, cognition, and emotion. When added to brain cellular cultures, anti-P antibodies caused a rapid and sustained increase in calcium influx in neurons, resulting in apoptotic cell death. In contrast, astrocytes, which do not express NSPA, were not affected. Injection of anti-P antibodies into the brain of living rats also triggered neuronal death by apoptosis. These results demonstrate a neuropathogenic potential of anti-P antibodies and contribute a mechanistic basis for psychiatric lupus. They also provide a molecular target for future exploration of this and other psychiatric diseases.

The carotid sinus connections: a WGA-HRP study in the cat

Previous neuroanatomical studies described the central representation of the carotid sinus nerve, but did not differentiate the projections of the baroreceptors from the chemoreceptors present in the carotid bifurcation. In this research we investigated the individual territories occupied by the primary afferents from the carotid body in the brainstem of the cat. We also studied the distribution of afferent and efferent neurons to the carotid body. We injected into the carotid body lectin coupled to horseradish peroxidase. We found labeled axons only in the nucleus of the tractus solitarius; in particular, we found strong projections to the following ipsilateral subnuclei: dorsal, interstitial, and medial part of the commissural subnucleus. Moderate labeling was found in the ipsilateral medial and intermediate subnuclei and in the contralateral dorsal subnucleus and the medial region of the commissural subnucleus. We found a mean of 256 +/- 79 (S.E.M.) labeled afferent ganglion cells in the petrosal ganglia, and no evidence of efferent neurons in the brainstem that innervate the carotid body; conversely, about 4000 efferent neurons of the superior cervical ganglion send terminals to the ipsilateral carotid body.

Maternal melatonin selectively inhibits cortisol production in the primate fetal adrenal gland

We tested the hypothesis that in primates, maternal melatonin restrains fetal and newborn adrenal cortisol production. A functional G‐protein‐coupled MT1 membrane‐bound melatonin receptor was detected in 90% gestation capuchin monkey fetal adrenals by (a) 2‐[125I] iodomelatonin binding (Kd, 75.7 ± 6.9 pm; Bmax, 2.6 ± 0.4 fmol (mg protein)−1), (b) cDNA identification, and (c) melatonin inhibition of adrenocorticotrophic hormone (ACTH)‐ and corticotrophin‐releasing hormone (CRH)‐stimulated cortisol but not of dehydroepiandrosterone sulphate (DHAS) production in vitro. Melatonin also inhibited ACTH‐induced 3β‐hydroxysteroid dehydrogenase mRNA expression. To assess the physiological relevance of these findings, we next studied the effect of chronic maternal melatonin suppression (induced by exposure to constant light during the last third of gestation) on maternal plasma oestradiol during gestation and on plasma cortisol concentration in the 4‐ to 6‐day‐old newborn. Constant light suppressed maternal melatonin without affecting maternal plasma oestradiol concentration, consistent with no effect on fetal DHAS, the precursor of maternal oestradiol. However, newborns from mothers under constant light condition had twice as much plasma cortisol as newborns from mothers maintained under a normal light–dark schedule. Newborns from mothers exposed to chronic constant light and daily melatonin replacement had normal plasma cortisol concentration. Our results support a role of maternal melatonin in fetal and neonatal primate cortisol regulation.

Arousal and differential Fos expression in histaminergic neurons of the ascending arousal system during a feeding-related motivated behaviour

Arousal depends on the concerted activity of the ascending arousal system (AAS) but specific stimuli may primarily activate some nuclei of this system. Motivated behaviours are characterized by behavioural arousal, although it is not known which AAS nuclei are active during a motivated behaviour. To address this issue, rats were rendered motivated for food by fasting them for 1 day and then were enticed with food that they could not obtain for varying periods of time. We studied the level of arousal by polysomnography or radiotelemetry, and Fos‐ir in the AAS, during food enticing. We found a strong arousal and an early increase in Fos‐ir in the histaminergic neurons from the tuberomammillary nucleus, after 30 min of enticing, followed by increased Fos‐ir in the whole AAS if food enticing was prolonged to 1 or 2 hours. In contrast, food presentation to non‐motivated rats did not increase arousal or Fos‐ir in the tuberomammillary nucleus. As opposed to the active arousal of the motivated rats, passive arousal induced by sensory stimulation was associated with increased Fos‐ir in the locus coeruleus and the orexin neurons, but not with increased Fos‐ir in the tuberomammillary nucleus or in the other nuclei of the AAS. We conclude that the arousal during feeding‐related motivated behaviour is associated primarily with the activation of the tuberomammillary nucleus, while the other arousal‐related nuclei become active later on.

The histamine H3 receptor and eating behavior.

Interest in the histaminergic system as a potential target for the treatment of feeding disorders is driven by the unsatisfactory history of the pharmacotherapy of obesity. Eating behavior is regulated by a complex interplay of central neurotransmitter systems, peripheral endocrine stimuli, the circadian rhythm, and environmental cues, all factors that change the behavioral state and alter homeostatic aspects of appetite and energy expenditure. Key factors driving eating behavior are appetite and satiety that are regulated through different mechanisms. Brain histamine has long been considered a satiety signal in the nervous system. Recent observations, however, indicate that histamine does not meet the criteria for being a satiety signal, because augmented histamine release accompanies the appetitive phase of feeding behavior rather than food consumption and satiety. The appetitive phase requires a high and yet optimal arousal state, and the histaminergic system is crucial for sustaining a high degree of arousal during motivated behavior. Histamine H1 receptors in the brain are crucial for the regulation of the diurnal rhythm of food intake and the regulation of obesity; however, from a therapeutic standpoint, no brain-penetrating H1 receptor agonists have been identified that would have antiobesity effects. Despite conflicting preclinical data, insights are emerging into the potential role of histamine H3 receptors as a target of antiobesity therapeutics. The aim of this review is to outline the relevance of the histaminergic system in controlling feeding behavior and evaluate the potential therapeutic use of histaminergic ligands for the treatment of eating disorders.

The circadian timing system: making sense of day/night gene expression.

The circadian time-keeping system ensures predictive adaptation of individuals to the reproducible 24-h day/night alternations of our planet by generating the 24-h (circadian) rhythms found in hormone release and cardiovascular, biophysical and behavioral functions, and others. In mammals, the master clock resides in the suprachiasmatic nucleus (SCN) of the hypothalamus. The molecular events determining the functional oscillation of the SCN neurons with a period of 24-h involve recurrent expression of several clock proteins that interact in complex transcription/translation feedback loops. In mammals, a glutamatergic monosynaptic pathway originating from the retina regulaltes the clock gene expression pattern in the SCN neurons, synchronizing them to the light:dark cycle. The emerging concept is that neural/humoral output signals from the SCN impinge upon peripheral clocks located in other areas of the brain, heart, lung, gastrointestinal tract, liver, kidney, fibroblasts, and most of the cell phenotypes, resulting in overt circadian rhythms in integrated physiological functions. Here we review the impact of day/night alternation on integrated physiology; the molecular mechanisms and input/output signaling pathways involved in SCN circadian function; the current concept of peripheral clocks; and the potential role of melatonin as a circadian neuroendocrine transducer.

Clock gene expression in adult primate suprachiasmatic nuclei and adrenal: is the adrenal a peripheral clock responsive to melatonin?

The circadian production of glucocorticoids involves the concerted action of several factors that eventually allow an adequate adaptation to the environment. Circadian rhythms are controlled by the circadian timing system that comprises peripheral oscillators and a central rhythm generator located in the suprachiasmatic nucleus (SCN) of the hypothalamus, driven by the self-regulatory interaction of a set of proteins encoded by genes named clock genes. Here we describe the phase relationship between the SCN and adrenal gland for the expression of selected core clock transcripts (Per-2, Bmal-1) in the adult capuchin monkey, a New World, diurnal nonhuman primate. In the SCN we found a higher expression of Bmal-1 during the h of darkness (2000-0200 h) and Per-2 during daytime h (1400 h). The adrenal gland expressed clock genes in oscillatory fashion, with higher values for Bmal-1 during the day (1400-2000 h), whereas Per-2 was higher at nighttime (about 0200 h), resulting in a 9- to 12-h antiphase pattern. In the adrenal gland, the oscillation of clock genes was accompanied by rhythmic expression of a functional output, the steroidogenic enzyme 3beta-hydroxysteroid dehydrogenase. Furthermore, we show that adrenal explants maintained oscillatory expression of Per-2 and Bmal-1 for at least 36 h in culture. The acrophase of both transcripts, but not its overall expression along the incubation, was blunted by 100 nm melatonin. Altogether, these results demonstrate oscillation of clock genes in the SCN and adrenal gland of a diurnal primate and support an oscillation of clock genes in the adrenal gland that may be modulated by the neurohormone melatonin.

The histaminergic tuberomammillary nucleus is critical for motivated arousal

Obtaining food, shelter or water, or finding a mating partner are examples of motivated behaviors, which are essential to preserve the species. The full expression of such behaviors requires a high but optimal arousal state. We tested the idea that tuberomammillary nucleus (TMN) histamine neurons are crucial to generate such motivated arousal, using a model of the appetitive phase of feeding behavior. Hungry rats enticed with food within a wire mesh box showed intense goal‐directed motor activity aimed at opening the box, an increase in core temperature, a fast histamine release in the hypothalamus and an early increase in Fos immunoreactivity in TMN and cortical neurons. Enticing with stronger‐tasting food induced stronger motor, temperature and Fos immunoreactivity brain responses than ordinary food pellets. TMN lesion greatly decreased all of those responses. We conclude that histamine neurons increase arousal and vegetative activity, allowing the normal unfolding of voluntary, goal‐directed behavior such as obtaining food.

Specific activation of histaminergic neurons during daily feeding anticipatory behavior in rats.

When food is available during a restricted and predictable time of the day, animals show increased locomotor and food searching behaviors before the anticipated daily meal. We had shown that histamine-containing neurons are the only aminergic neurons related to arousal that become active in anticipation of an upcoming meal. To further map, the brain regions involved in the expression of the feeding-anticipatory behavior, we quantified the expression of Fos in hypothalamic areas involved in arousal. We found that nearly 35% of the histamine neurons from the tuberomammillary nucleus were Fos-immunoreactive immediately before mealtime. One hour before this transient increase in Fos-immunoreactivity, we found a similarly brief increase of fos mRNA in the tuberomammillary nucleus. In contrast, the activation of two types of perifornical hypothalamic neurons followed meal onset by 1-2 h. One neuron type was orexin/hypocretin-immunoreactive, while the other type was neither orexin nor melanin concentrating hormone-immunoreactive. The present work indicates that the increased locomotor activity that anticipates mealtime coincides with the activation of the tuberomammillary nucleus, and that the behavioral activation during the consummatory phase of feeding coincides more closely with the delayed activation of the perifornical hypothalamic area.

Ultrastructure of glutamate and GABA immunoreactive axon terminals of the rat nucleus tractus solitarius, with a note on infralimbic cortex afferents

The principal fast neurotransmitters in the CNS are glutamate and GABA. Our aim was to provide a baseline account on the ultrastructure of the axon terminals immunoreactive to glutamate or GABA present in the nucleus tractus solitarius (NTS) of the rat. In addition, we wanted to complete our study of cortico-solitary afferents at the electron microscopic level, by analyzing the inputs from the infralimbic cortex. Using post-embedding immunogold, we found that nearly 61% of the axon terminals were glutamatergic, and 36% were GABAergic in the rat visceral NTS. In general, axons making asymmetric synaptic contacts were enriched in glutamate, compared to axons involved in symmetric synapses. In contrast, the vast majority of the GABAergic axon terminals made symmetric synaptic contacts. We could discern five types of glutamatergic and two types of GABAergic axon terminals that differed in their fine structure. Afferents from the infralimbic cortex were small, with clear synaptic vesicles and no dense core vesicles; they made asymmetric contacts with fine dendrites, and were glutamatergic. We conclude that most axon terminals in the NTS use glutamate or GABA as fast transmitters, in addition to being a heterogeneous population of morphological types.