Distribution of neuropeptide Y-like immunoreactivity in the rat central nervous system--I. Radioimmunoassay and chromatographic characterisation |
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Authors: | M E de Quidt P C Emson |
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Institution: | 1. Department of Neurology, The Affiliated Hospital of the Qingdao University, Qingdao, Shandong, 266100, China;2. Department of Emergency, The Affiliated Hospital of the Qingdao University, Qingdao, Shandong, 266100, China;1. Section of Hematology/Oncology, Department of Medicine, University of Chicago, Chicago, IL, USA;2. Biological Sciences Collegiate Division, University of Chicago, Chicago, IL, USA;3. Rush Medical College, Rush University Medical Center, Chicago, IL, USA;4. School of Medicine, Stanford University, Stanford, CA, USA;1. Laboratory of Neuroendocrinology & Nutritional and climatic Environment, University Sidi Mohamed Ben Abdellah, Faculty of Sciences DM, Fez, Morocco;2. INSERM U982, Laboratory of Neuronal and Neuroendocrine Differentiation and Communication, Rouen University, Institute for Research and Innovation in Biomedicine (IRIB), 76821 Mont-Saint-Aignan, France;3. Normandy University, Caen, France;1. Neurology Unit, Department of Clinical and Experimental Sciences, University of Brescia, Italy;2. Stroke Unit, Azienda Ospedaliera “Spedali Civili”, Spedali Civili Hospital, Brescia, Italy;3. Department of Neurodegeneration and Hertie Institute for Clinical Brain Research, University of Tuebingen, Germany;4. Department of Medical Imaging, Geneva University Hospital, Geneva, Switzerland;5. Nuclear Medicine Unit, Azienda Azienda Ospedaliera “Spedali Civili”, Spedali Civili Hospital, Brescia, Italy;1. Laboratory of Veterinary Pathology, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu-shi, Tokyo 183-8509, Japan;2. Pathogenetic Veterinary Science, United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan;3. Laboratory of Food Safety Sciences, Azabu University, 1-17-71, Fuchinobe, Chuo-ku, Sagamihara-shi, Kanagawa 252-0206, Japan |
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Abstract: | The distribution of neuropeptide Y-like immunoreactivity in the rat brain was investigated by means of immunochemical techniques. In the first part of the study (present paper) neuropeptide Y radioimmunoassays were characterised and the chromatographic properties and regional distribution of neuropeptide Y-like immunoreactivity was investigated. The second part of the study (accompanying paper) involved immunohistochemical techniques. Extracts from several regions of rat brain were found to contain immunoreactivity that behaved like synthetic porcine neuropeptide Y in three test systems: dilution in the radioimmunoassay (test of antigenic properties), gel chromatography (molecular weight), reverse phase high performance liquid chromatography (solubility properties). Experiments were conducted to optimise the extraction of neuropeptide Y. Boiling 0.1 M sodium phosphate buffer, pH 7.4, extracted at least two times as much immunoreactivity from whole brain pieces as other buffers. The nature of the extracted immunoreactivity was confirmed using chromatography. Experiments (using added iodinated or unlabelled neuropeptide Y standards) demonstrated that the differences between extraction media could not be explained by differential recovery of the peptide, although differences in recovery between media existed. Tissue sample weight was found to influence neuropeptide Y recovery. Evidence that rat neuropeptide Y-like immunoreactivity was not identical to the porcine peptide was obtained from experiments which demonstrated an early eluting peak of immunoreactivity in addition to the main peak on high performance liquid chromatograms. This material could be generated by oxidation of extracted rat neuropeptide Y, suggesting the presence in the rat peptide of a methionine residue. Some evidence of high molecular weight neuropeptide Y precursors was obtained from chromatography of hypothalamus extracts. Bovine pancreatic polypeptide-like material represented less than 1% of the amounts of neuropeptide Y in the brain. The distribution of neuropeptide Y-like immunoreactivity was non-uniform in the rat brain with highest concentrations observed in the hypothalamus, amygdaloid complex and periaqueductal central gray matter. Other regions of forebrain contained moderate to high concentrations including olfactory tubercle, striatum, nucleus accumbens, neocortex and hippocampus. Negligible amounts were detected in the cerebellum. In spinal cord immunoreactivity was concentrated in the dorsal horn, although measurable amounts were found in the ventral horn. The neurointermediate but not anterior lobe of the pituitary contained neuropeptide Y. |
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