RESUMEN
We tested the hypothesis that, in the amphibian Xenopus laevis, cocaine- and amphetamine-regulated transcript peptide (CARTp) not only has widespread actions in the brain but also acts as a local factor in endocrine pituitary cells and/or is neurohemally secreted into the circulation to control peripheral targets. CARTp-immunoreactive cells occur in the olfactory bulb, nucleus accumbens, amygdala, septum, striatum, nucleus of Bellonci, ventrolateral nucleus, central thalamic nucleus, preoptic nuclei, and suprachiasmatic nucleus, and particularly in the medial pallium, ventromedial nucleus, hypothalamus, Edinger-Westphal nucleus, optic tectum, raphe nuclei, central gray, nucleus of the solitary tract, and spinal cord. From the hypothalamic magnocellular nucleus, CARTp-containing axons run to the neurohemal median eminence, and to the neural pituitary lobe to form neurohemal terminals, as shown by immunoelectron microscopy. Starvation increases the number of CARTp-cells in the optic tectum by 46% but has no effect on such cells in the torus semicircularis. CARTp does not affect in vitro release of alpha-melanophore-stimulating hormone from pituitary melanotrope cells. Our results support the hypothesis that in X. laevis, CARTp not only has multiple and not exclusively feeding-related actions in the brain but is also secreted as a neurohormone 1) into the portal system to control endocrine targets in the pituitary distal lobe and 2) from neurohemal axon terminals in the neural pituitary lobe to act peripherally. The differences in CARTp distribution between X. laevis and Rana esculenta may be related to different environmental and physiological conditions such as feeding, sensory information processing, and locomotion.
Asunto(s)
Encéfalo/metabolismo , Proteínas del Tejido Nervioso/biosíntesis , Hipófisis/metabolismo , Xenopus laevis/metabolismo , Animales , Conducta Alimentaria/fisiología , Inmunohistoquímica , Microscopía Inmunoelectrónica , Médula Espinal/metabolismoRESUMEN
The presence of the opioids, beta-endorphin, met-enkephalin, and endomorphin, and of corticotropin-releasing factor (CRF) and the CRF family member, urocortin (Ucn), is described in cerebrospinal fluid-contacting neurons in the brain of the amphibian, Xenopus laevis.
Asunto(s)
Hormona Liberadora de Corticotropina/líquido cefalorraquídeo , Neuronas/metabolismo , Péptidos Opioides/líquido cefalorraquídeo , Xenopus laevis/líquido cefalorraquídeo , Animales , UrocortinasRESUMEN
Brain-derived neurotrophic factor (BDNF) is immunocytochemically demonstrated in the magnocellular nucleus and neural lobe of the pituitary gland of the amphibian Xenopus laevis. Immunoelectron microscopy shows BDNF in secretory granules of type A neurohemal axon terminals in the neural lobe of pituitary gland. It is proposed that BDNF released from the neural lobe acts as a neurohormone stimulating the secretory activity of the melanotrope cells in the intermediate pituitary lobe.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/fisiología , Sistema Hipotálamo-Hipofisario/química , Proteínas de Xenopus/fisiología , Animales , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Sistema Hipotálamo-Hipofisario/metabolismo , Sistema Hipotálamo-Hipofisario/ultraestructura , Hipófisis/química , Hipófisis/metabolismo , Hipófisis/ultraestructura , Proteínas de Xenopus/metabolismo , Xenopus laevisRESUMEN
We have raised the hypothesis that in the South African clawed toad Xenopus laevis, urocortin 1 (UCN1), a member of the corticotropin-releasing factor (CRF) peptide family, functions not only within the brain as a neurotransmitter/neuromodulator but also as a neurohormone, promoting the release of alpha-melanophore-stimulating hormone (alphaMSH) from the neuroendocrine melanotrope cells in the intermediate lobe of the pituitary gland. This hypothesis has been investigated by (1) assessing the distribution of UCN1 and CRF by light immunocytochemistry, (2) determining the subcellular presence of UCN1 in the neural lobe of the pituitary gland by immuno-electron microscopy applying high-pressure freezing and cryosubstitution, and (3) testing the effect of UCN1 on MSH release from toad melanotrope cells using in vitro superfusion. In the X. laevis brain, the main site of UCN1-positive somata was found to be the Edinger-Westphal nucleus. UCN1 immunoreactivity (ir) also occurs in the nucleus posteroventralis tegmenti, central gray, nucleus reticularis medius, nucleus motorius nervi facialis, and nucleus motorius nervi vagi. UCN1 occurs together with CRF in the nucleus motorius nervi trigemini, and in the magnocellular nucleus, which send a UCN1- and CRF-containing fiber tract to the median eminence. Strong UCN1-ir and CRF-ir were found in the external zone of the median eminence. From the internal zone of the median eminence, UCN1-ir fibers, but few CRF-ir fibers, were found to project to the pituitary neural lobe, where they form numerous neurohemal axon terminals. Ultrastructurally, two types of terminal containing UCN1-ir secretory granules were distinguished: type A contains large, moderately electron-dense, round secretory granules and type B is filled with smaller, strongly electron-dense, ellipsoid secretory granules. In vitro superfusion studies showed that UCN1 stimulated the release of alphaMSH from melanotrope cells in a dose-dependent manner. Our results support the hypothesis that in X. laevis, UCN1 released from neurohemal axon terminals in the pituitary neural lobe functions as a stimulatory neurohormone for alphaMSH release from melanotrope cells of the pituitary intermediate lobe.
Asunto(s)
Hormona Liberadora de Corticotropina/metabolismo , Hipófisis/metabolismo , alfa-MSH/metabolismo , Animales , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Hormona Liberadora de Corticotropina/farmacología , Técnicas In Vitro , Neurotransmisores/metabolismo , Hipófisis/efectos de los fármacos , Hormonas Hipofisarias/metabolismo , Urocortinas , Xenopus laevisRESUMEN
The distribution of cocaine- and amphetamine-regulated transcript peptide (CARTp)- like immunoreactivity was studied only in the rat central nervous system (CNS). In mammals, CART peptides occur among others in brain areas that control feeding behavior. We mapped CARTp-immunoreactive structures in the CNS of the frog Rana esculenta and assumed that differences may exist in the CARTp-containing neuronal populations between the frog, which does not feed in winter, and the rat. In the forebrain, immunoreactive cells and fibers were found in the olfactory bulb, nucleus accumbens, amygdala, medial pallium, septum, striatum, the preoptic nuclei, ventromedial nucleus, central thalamic nucleus, and the hypothalamus. The optic pathway was free of immunoreactivity. The neurohypophysis showed intense immunostaining. In the mesencephalon, many cells were stained in the Edinger-Westphal nucleus, and a few in the optic tectum, where fibers were stained in all plexiform layers. In the retina, some cells in the inner nuclear layer contained CARTp. In the rhombencephalon, cells were stained in the raphe nuclei, central gray, nucleus of the solitary tract, and the vicinity of motor nuclei. Neurons of the motor cranial nerves were densely innervated by CARTp-positive fibers originating from the spinal cord. In the spinal cord, preganglionic cells were stained, and motoneurons were surrounded by immunoreactive varicose axon terminals. Major differences were found between the frog and the rat brains in the distribution of CARTp in the visual system, olfactory bulb, preoptic area, and the motor nuclei. Some of these differences may be related to feeding behavior of these animals.
Asunto(s)
Sistema Nervioso Central/metabolismo , Inmunohistoquímica/métodos , Proteínas del Tejido Nervioso/metabolismo , Neuronas/metabolismo , Animales , Western Blotting/métodos , Sistema Nervioso Central/citología , Rana esculentaRESUMEN
In secretory cells filamentous actin (f-actin) is mostly present subjacent to the plasma membrane, referred to as cortical actin. While the function of cortical actin in the secretory processes has been extensively studied, little attention has been given to the role of actin in signal transduction and intracellular second messenger dynamics. Analysis with the fluorescent f-actin probe Alexa-phalloidin shows that Xenopus laevis pituitary melanotrope cells possess a thick cortical actin ring. This cell is a good model to study the possible function(s) of f-actin in signal transduction processes. Regulation of the release of alpha-MSH from this cell involves a convergence of various receptor mechanisms to regulate the activity of voltage-operated Ca2+ channels. We have considered three potential functions for the cortical actin ring in the signaling process of the melanotrope: (1) it functions as a barrier for access of secretory granules to the membrane for exocytosis, (2) it is involved in anchoring components of the Ca2+ signalling machinery of the cell, and/or (3) it helps to form a scaffold for components of the signal transduction machinery used by the various neurotransmitters and neuropeptides that regulate the activity of the cell. To test these possibilities we have examined the effect of the f-actin depolymerising toxin latrunculin B on Ca2+ signaling, signal transduction and alpha-MSH secretion in the melanotrope. We show that while the toxin is effective in disrupting the cortical actin ring, this treatment has no effect on either Ca2+ signaling or the signal transduction processes studied. The toxin does induce an increase in alpha-MSH release, indicating that the cortical actin ring acts as a barrier for secretory granule access to the membrane.