Containing Corticotropin-releasing Factor Research Articles

Corticotropin-Releasing Factor Release From a Unique Subpopulation of Accumbal Neurons Constrains Action-Outcome Acquisition in Reward Learning

BackgroundThe nucleus accumbens (NAc) mediates reward learning and motivation. Despite an abundance of neuropeptides, peptidergic neurotransmission from the NAc has not been integrated into current models of reward learning. The existence of a sparse population of neurons containing corticotropin-releasing factor (CRF) has been previously documented. Here, we provide a comprehensive analysis of their identity and functional role in shaping reward learning. MethodsOur multidisciplinary approach included fluorescent in situ hybridization (n = ≥3 mice), tract tracing (n = 5 mice), ex vivo electrophysiology (n = ≥30 cells), in vivo calcium imaging with fiber photometry (n = ≥4 mice), and use of viral strategies in transgenic lines to selectively delete CRF peptide from NAc neurons (n = ≥4 mice). Behaviors used were instrumental learning, sucrose preference, and spontaneous exploration in an open field. ResultsWe showed that the vast majority of NAc CRF-containing neurons are spiny projection neurons (SPNs) comprising dopamine D1-, D2-, or D1/D2-containing SPNs that primarily project and connect to the ventral pallidum and to a lesser extent the ventral midbrain. As a population, they display mature and immature SPN firing properties. We demonstrated that NAc CRF-containing neurons track reward outcomes during operant reward learning and that CRF release from these neurons acts to constrain initial acquisition of action-outcome learning and at the same time facilitates flexibility in the face of changing contingencies. ConclusionsCRF release from this sparse population of SPNs is critical for reward learning under normal conditions.

CRF release from a unique subpopulation of accumbal neurons constrains action-outcome acquisition in reward learning.

The nucleus accumbens (NAc) mediates reward learning and motivation. Despite an abundance of neuropeptides, peptidergic neurotransmission from the NAc has not been integrated into current models of reward learning. The existence of a sparse population of neurons containing corticotropin releasing factor (CRF) has been previously documented. Here we provide a comprehensive analysis of their identity and functional role in shaping reward learning. To do this, we took a multidisciplinary approach that included florescent in situ hybridization (N mice ≥ 3), tract tracing (N mice = 5), ex vivo electrophysiology (N cells ≥ 30), in vivo calcium imaging with fiber photometry (N mice ≥ 4) and use of viral strategies in transgenic lines to selectively delete CRF peptide from NAc neurons (N mice ≥ 4). Behaviors used were instrumental learning, sucrose preference and spontaneous exploration in an open field. Here we show that the vast majority of NAc CRF-containing (NAc CRF ) neurons are spiny projection neurons (SPNs) comprised of dopamine D1-, D2- or D1/D2-containing SPNs that primarily project and connect to the ventral pallidum and to a lesser extent the ventral midbrain. As a population, they display mature and immature SPN firing properties. We demonstrate that NAc CRF neurons track reward outcomes during operant reward learning and that CRF release from these neurons acts to constrain initial acquisition of action-outcome learning, and at the same time facilitates flexibility in the face of changing contingencies. We conclude that CRF release from this sparse population of SPNs is critical for reward learning under normal conditions.

Localization of corticotropin-releasing factor immunoreactivity in the brain of the teleost Sparus aurata

The distribution of perikarya and fibers containing corticotropin-releasing factor (CRF) was studied in the brain of the teleost Sparus aurata by immunocytochemistry using the peroxidase-antiperoxidase method. Antisera against rat CRF, arginine vasotocin, and human adrenocorticotropin (ACTH) were used. Most CRF-immunoreactive neurons were located in the nucleus lateralis tuberis, but they were absent from the nucleus preopticus, which only contained arginine vasotocin neurons. Few CRF perikarya were identified in the nucleus preopticus periventricularis and in the mesencephalic tegmentum. A conspicuous bundle of immunoreactive fibers ran along the diencephalic floor and pituitary stalk to end near the cells of the hypophysial pars intermedia. No CRF was seen near the adenohypophysial rostral pars distalis. Our results suggest that, in Sparus aurata, CRF is a releasing factor for melanotropic cells. Its role as a releasing factor for ACTH is discussed.

Influence of Immune Signals on the Hypothalamic-Pituitary Axis of the Rodent

The immune system and the hypothalamic-pituitary (H-P) axis are functionally connected, so that exposure to antigens elicits a coordinated response which allows the organism to successfully withstand immunologic challenges. An important feature of this bilateral communication is the appearance of proteins released into the circulation by activated immune cells. These proteins, called cytokines or interleukins, stimulate the activity of the H-P axis, thus increasing circulating ACTH and corticosteroid levels. This in turn induces metabolic changes such as increases in energy substrates, restrains the activity of specific immune cells, and alters the release of secretagogues important for both the immune system and neuroendocrine organs. After acute increases in blood-borne levels of cytokines, nerve terminals in the median eminence, particularly those containing corticotropin-releasing factor (CRF), represent an important site of action of these immune signals. Subsequently, changes take place within the brain in general and the hypothalamus in particular. They include the synthesis/release of peptides such as CRF and vasopressin, of neurotransmitters such as catecholamines and prostaglandins, and of cytokines themselves. Upon prolonged exposure to blood-borne immune signals additional mechanisms come into play, including those taking place directly at the pituitary level. These observations indicate that cytokines released into the general circulation act on multiple sites within the H-P axis, a phenomenon that relies on the recruitment of a large number of pathways. This review discusses these pathways and the mechanisms through which they allow cytokines to convey the occurrence of immune activation to the brain.

Distribution of corticotropin-releasing factor immunoreactive neurons in the brain of the tigerfrog, Rana tigrina

The distribution of immunoreactive (ir) neurons containing corticotropin-releasing factor (CRF) is described in the brain of the tigerfrog, Rana tigrina. The olfactory bulb, medial pallium, nucleus of the diagonal band of Broca and medial area of the amygdala of the telencephalon showed ir-CRF perikarya. The anterior and ventromedial thalamic nuclei, and the magnocellular nucleus preopticus (NPO) revealed several ir cells; a few NPO neurons were cerebrospinal fluid contacting in nature. The nucleus hypothalamicus ventromedialis contained a few cells, but the nucleus infundibularis ventralis of the infundibulum revealed several diffusely distributed perikarya. Individual ir-CRF perikarya were visualized in the optic tectum and interpeduncular nucleus. Extensive fiber terminals were present in the median eminence, but no fibers were discerned either in the neural lobe or in the pituitary gland.

Synaptic associations between oxytocin-containing magnocellular neurons and neurons containing corticotropin-releasing factor in the rat magnocellular paraventricular nucleus

In the paraventricular nucleus (PVN) of the rat hypothalamus, we determined synaptic associations between oxytocin (OXT)-containing magnocellular neurons and parvocellular neurons containing corticotropin-releasing factor (CRF) by using a double immunolabeling technique in 7 animals. In single vibratome sections of the hypothalamus, immunoreactive CRF and OXT were labeled with silver-gold particles and diaminobenzidine (DAB) chromogen, respectively. By light microscopy CRF-containing fibers appeared to be black dots, some of which encircled magnocellular perikarya labeled with brown DAB chromogen in the PVN. By electron microscopy we discriminated OXT neurons having fine DAB-chromogen throughout the cytoplasm and on large secretory granules from CRF neurons having dense coarse particles of silver-gold. Occasional CRF axons terminated on perikarya or dendritic processes of OXT neurons, making synaptic contacts. The terminals which were characterized by having clusters of small clear vesicles and a few dense core vesicles showed equal thickenings of pre- and postsynaptic membranes at the synaptic junctions.

The distribution of corticotropin-releasing factor-immunoreactive neurons and nerve fibers in the brain of the snake, Natrix maura

The anatomical distribution of neurons and nerve fibers containing corticotropin-releasing factor (CRF) has been studied in the brain of the snake, Natrix maura, by means of immunocytochemistry using an antiserum against rat CRF. To test the possible coexistence of CRF with the neurohypophysial peptides arginine vasotocin (AVT) and mesotocin (MST) adjacent sections were stained with antisera against the two latter peptides. CRF-immunoreactive (CRF-IR) neurons exist in the paraventricular nucleus (PVN). In some neurons of the PVN, coexistence of CRF with MST or of CRF with AVT has been shown. Numerous CRF-IR fibers run along the hypothalamo-hypophysial tract and end in the outer layer of the median eminence. In addition, some fibers reach the neural lobe of the hypophysis. CRF-IR perikarya have also been identified in the following locations: dorsal cortex, nucleus accumbens, amygdala, subfornical organ, lamina terminalis, nucleus of the paraventricular organ, nucleus of the oculomotor nerve, nucleus of the trigeminal nerve, and reticular formation. In addition to all these locations CRF-IR fibers were also observed in the lateral septum, supraoptic nucleus, habenula, lateral forebrain bundle, paraventricular organ, hypothalamic ventromedial nucleus, raphe and interpeduncular nuclei.

Influence of the central nucleus of the amygdala on the content of corticotropin-releasing factor in the median eminence.

The central nucleus of the amygdala possesses numerous neurons containing corticotropin-releasing factor (CRF). This study demonstrates a striking decrease of the CRF-like immunoreactivity in the median eminence at both 1 and 2 weeks after bilateral lesions of the amygdaloid central nucleus. Lesion of the amygdaloid central nucleus did not alter the neurophysin-like immunoreactivity in the internal zone of the median eminence, indicating the integrity of the efferent neurophysin-containing fibers of the supraoptic and paraventricular hypothalamic nuclei. However, there was a concomitant decrease of neurophysin and CRF-like immunoreactivity in the external zone of the median eminence. These results substantiate the hypothesis that the amygdaloid central nucleus can influence the content of CRF-like material in the median eminence via a multisynaptic pathway involving the synthesis of CRF at the level of the paraventricular nucleus of the hypothalamus. The exact mechanism by which lesion of the amygdaloid central nucleus influences the CRF content in the median eminence remains to be determined.

Corticotropin-releasing factor-like immunoreactive nerve fibers in the rat superior cervical ganglion and their fine structures

The existence of nerve fibers containing corticotropin-releasing factor (CRF)-like immunoreactivity (CRFI) in the rat superior cervical ganglion (SCG) was demonstrated by using immunocytochemistry. They were found to be extrinsic in origin, because no CRFI neurons were seen in the SCG and decentralization resulted in the disappearance of CRFI fibers in the SCG on the operated side. These findings were also confirmed by immunoelectron microscopic analysis; CRFI fibers contained a number of small clear synaptic vesicles but were devoid of large granular and agranular vesicles. These morphological characteristics are identical to those of the preganglionic fibers. The present immunoelectron microscopic analysis revealed that most of the CRFI fibers in the SCG make synaptic contact predominantly with the dendrites of the principal cells, partly with their somas and rarely with a no-n CRFI terminal. Thus, the present study provides direct morphological evidence that CRF directly influences the function of the principal cells of the SCG and that CRFI fibers are preganglionic.

Catecholaminergic Innervation of Neurons Containing Corticotropin-Releasing Factor in the Paraventricular Nucleus of the Rat Hypothalamus

Catecholaminergic synaptic input to neurons containing corticotropin-releasing factor (CRF) in the parvocellular portion of the paraventricular nucleus (PVN) in the rat hypothalamus was observed. The experimental techniques used combine autoradiography after 3H-noradrenaline (3H-NA) injection or uptake of 5-hydroxydopamine (5-OHDA) with immunocytochemistry using CRF antiserum. CRF-like immunoreactive cell bodies and fibers in the PVN received synaptic inputs from the axon terminals in which a selective accumulation of 3H-NA or 5-OHDA was found. This finding suggests that the secretion of CRF neurons may be regulated via synapses by catecholaminergic neurons.

Radioautographic study of binding and internalization of corticotropin-releasing factor by rat anterior pituitary corticotrophs.

In order to identify the anterior pituitary cell type(s) containing corticotropin-releasing factor (CRF) receptors and to study the internalization processes of this peptide by the target cells, radioautography was performed on rat anterior pituitaries removed at specific intervals (2-60 min) after intracarotid injection of [125I]iodo-CRF into intact and adrenalectomized female rats. In intact animals, all corticotrophs were labeled, whereas in the adrenalectomized animals about 80% of the hypertrophied corticotrophs (adrenalectomy cells) were. In control animals injected with both iodinated CRF and an excess of unlabeled peptide, no specific reaction could be detected. The time-course study in intact animals showed that 2 min after injection most silver grains were found over or within 160 nm of the plasma membrane. At the 5-min time intervals, grains were observed both over the plasma membrane and within the cytoplasm, associated with lysosomes, and the Golgi apparatus. Fifteen minutes after injection, grains were mostly found over lysosomes and the Golgi apparatus, whereas at the longest time intervals (30 and 60 min) almost no labeling could be detected. The results obtained in this study indicate that in the anterior pituitary CRF receptors are restricted to corticotrophs (as identified by electron microscopy) and that, after binding to the plasma membrane, CRF is rapidly internalized to Golgi elements and lysosomes.

Differences in the distributional pattern of CRF-, oxytocin-, and vasopressin-immunoreactive nerve fibers in the median eminence of the rat.

Conspicuous differences in the distributional pattern of nerve fibers containing corticotropin-releasing factor (CRF) or posterior lobe hormones, respectively, were shown in the median eminence of the adult male rat by means of immunoperoxidase histochemistry, with the use of anti-CRF, anti-oxytocin, and anti-vasopressin sera. In the rostral and central divisions of the median eminence, a high concentration of CRF-immunoreactive nerve fibers was found in the median portion of the external layer; these fibers terminated on the capillary loops of the hypophysial portal system. In the caudal division of the median eminence, the CRF-immunoreactive nerve fibers were located in the median to paramedian portions of the external layer. Numerous oxytocin- and vasopressin-immunoreactive nerve fibers were observed evenly distributed throughout the internal layer of the median eminence. In the external layer, a small number of the oxytocin- and vasopressin-containing nerve fibers was found around the capillary loops, particularly in the median to paramedian portions. The distributional patterns of the CRF and the posterior lobe hormones in the hypothalamo-hypophysial system and their functional interrelationship are discussed.