ABSTRACT
The blood-brain barrier (BBB) plays an important role in controlling the access of substances to the brain. Of the circumventricular organs (CVO), i.e. areas that lack a BBB, the median eminence and its close relationship with the hypothalamic arcuate nucleus plays an important role in controlling the entry of blood-borne substances to neurons of the mediobasal hypothalamus. In order to clarify the nature of the BBB in the median eminence-arcuate nucleus complex, we have used immunohistochemistry and antisera to protein components of the BBB-(1) tight junctions, claudin-5 and zona occludens-1 (ZO-1); (2) endothelial cells: (a) all endothelial cells: rat endothelial cell antigen-1 (RECA-1), (b) endothelial cells at BBB: endothelial barrier antigen (EBA), glucose transporter 1 (GLUT1) and transferrin receptor (TfR), and (c) endothelial cells at CVOs: dysferlin; (3) basal lamina: laminin; (4) vascular smooth muscle cells: smooth muscle actin (SMA); (5) pericytes: chondroitin sulfate proteoglycan (NG2); (6) glial cells: (a) astrocytes: glial fibrillary acidic protein (GFAP), (b) tanycytes: dopamine- and cAMP-regulated phosphoprotein of 32kDA (DARPP-32), (c) microglia: CD11b. Neuronal cell bodies located in the ventromedial aspect of the arcuate nucleus were visualized by antiserum to agouti-related protein (AgRP). The study provides a detailed analysis on the cellular localization of BBB components in the mediobasal hypothalamus. Some vessels in the ventromedial aspect of the arcuate nucleus lacked the BBB markers EBA and TfR, suggesting an absence of an intact BBB. These vessels may represent a route of entry for circulating substances to a subpopulation of arcuate nucleus neurons.
Subject(s)
Blood-Brain Barrier/metabolism , Endothelial Cells/metabolism , Hypothalamus/blood supply , Hypothalamus/metabolism , Microcirculation/metabolism , Tight Junctions/metabolism , Animals , Arcuate Nucleus of Hypothalamus/blood supply , Arcuate Nucleus of Hypothalamus/metabolism , Arcuate Nucleus of Hypothalamus/ultrastructure , Biomarkers/metabolism , Blood-Brain Barrier/ultrastructure , Claudin-5 , Endothelial Cells/ultrastructure , Hypothalamus/ultrastructure , Immunohistochemistry , Male , Median Eminence/blood supply , Median Eminence/metabolism , Median Eminence/ultrastructure , Membrane Proteins/metabolism , Microcirculation/ultrastructure , Myocytes, Smooth Muscle/metabolism , Myocytes, Smooth Muscle/ultrastructure , Nerve Tissue Proteins/metabolism , Neuroglia/metabolism , Neuroglia/ultrastructure , Neurons/metabolism , Neurons/ultrastructure , Pericytes/metabolism , Pericytes/ultrastructure , Phosphoproteins/metabolism , Rats , Rats, Sprague-Dawley , Tight Junctions/ultrastructure , Zonula Occludens-1 ProteinABSTRACT
Several lines of evidence support a role for pituitary adenylate cyclase-activating polypeptide (PACAP) in the regulation of energy balance. In the present study, we have used fluorescent in situ hybridization and immunohistochemistry to investigate in detail the cellular localization and chemical content of PACAP mRNA- and peptide-containing neuronal cell bodies in the mediobasal hypothalamus of the rat. PACAP mRNA-containing cell bodies were demonstrated in high numbers in the ventromedial hypothalamic nucleus (VMH) and in lower numbers in the arcuate nucleus (Arc). In colchicine-treated rats, PACAP immunoreactivity was demonstrated in many cell bodies of the VMH and several cell bodies of the ARC. Double-labeling revealed that PACAP immunoreactivity was present in approximately 20% of pro-opiomelanocortin (POMC) neurons in the ventrolateral Arc as shown by presence of alpha-melanocyte-stimulating hormone (alpha-MSH), but not in agouti-related peptide (AgRP)-containing neurons in the ventromedial aspect of the Arc. PACAP immunoreactivity was also colocalized with the vesicular acetylcholine transporter (VAChT; a marker for cholinergic neurons) in Arc POMC neurons. Brainstem POMC neurons in the commissural part of the solitary tract nucleus were devoid of PACAP immunoreactivity. However, several VAChT-positive neurons in the dorsal motor nucleus of the vagus nerve were also PACAP immunoreactive, whereas VAChT-positive neurons of the motor nucleus of the hypoglossal nerve were PACAP-negative. The results show presence of PACAP with alpha-MSH in a subpopulation of hypothalamic POMC neurons and point further to the neurochemical heterogeneity of hypothalamic, but not brainstem, POMC neurons.
Subject(s)
Arcuate Nucleus of Hypothalamus/metabolism , Neurons/metabolism , Pituitary Adenylate Cyclase-Activating Polypeptide/metabolism , Pro-Opiomelanocortin/metabolism , alpha-MSH/metabolism , Agouti-Related Protein/metabolism , Animals , Arcuate Nucleus of Hypothalamus/cytology , Energy Metabolism/physiology , Immunohistochemistry , Male , Pituitary Adenylate Cyclase-Activating Polypeptide/genetics , RNA, Messenger/analysis , Rats , Rats, Sprague-Dawley , Tissue Distribution , Ventromedial Hypothalamic Nucleus/cytology , Ventromedial Hypothalamic Nucleus/metabolism , Vesicular Acetylcholine Transport Proteins/metabolismABSTRACT
Endogenous opioid signaling contributes to the neural control of food intake. Opioid signaling is thought to regulate palatability, the reward value of a food item as determined by orosensory cues such as taste and texture. The reward value of a food reflects not only these sensory properties but also the relative value of competing food choices. In the present experiment, we used a consummatory contrast paradigm to manipulate the relative value of a sucrose solution for two groups of rats. Systemic injection of the nonspecific opioid antagonist naltrexone suppressed sucrose intake; for both groups, however, this suppression was selective, occurring only for the relatively more valuable sucrose solution. Our results indicate that endogenous opioid signaling contributes to the encoding of relative reward value.
