Protein HMG-17 is hyper-expressed in rat glucagonoma. Single-step isolation and sequencing

High-mobility-group protein 17 (HMG-17) was identified by reversed-phase high-performance liquid chromatography analysis as a major component in acidic extracts of transplantable rat glucagonoma tissue but not in insulinoma tissue of similar origin. The peptide was purified in a single step and the entire sequence of 89 amino acids was determined. Rat HMG-17 has a molecular mass of 9238 Da and shows strong similarity to human, bovine (94.4%) and chicken (88.8%) HMG-17. Six of the seven residues which vary among the mammalian sequences are located within a short segment (positions 64-83) present in the acidic, non-DNA-binding C-terminal part of HMG-17. This region shows least similarity to the otherwise related proteins HMG-14 and H6 (a trout HMG protein). Interestingly, four of the six variable positions are Asp in rat HMG-17 which results in an overall net increase in the negative charge of the C-terminal region. The nature of selective hyper-expression of HMG-17 in glucagon but not in insulin-producing tumor tissue remains to be clarified.     

Localisation of oxytocin,vasopressin and parts of precursors in the human neonatal adrenal

Summary Being a possible alternative source for the production of vasopressin (AVP) and oxytocin (OXT), a study was undertaken of the fetal adrenal. The concentrations of these peptides within the fetal adrenal turned out to be low, viz., approx. 1 pg/mg in the rat and within the pg/g range in the human. Immunocytochemistry was performed either on conventional autopsy material kept till 12 years in paraffin blocks, or on more recently obtained formalin or glutaraldehyde-paraformaldehyde fixed material. In both types of material staining was good. In order to localize AVP cells, anti-AVP, an antibody against its associated neurophysin (anti-NSN) or an antibody raised against the c-terminal glycopeptide part of the AVP precursor (anti-GP) was used. OXT cells were localized by means of anti-OXT or an auto-antibody of a multiple sclerosis patient (auto-MS) probably recognizing OXT-neurophysin. The antibodies were characterized on human and rat brain material. In the external zone of the definitive cortex, apart from parenchyma cells, anti-AVP, anti-NSN and anti-GP stained fibre-like structures running in the connective tissue septa and around parenchyma cells and the cytoplasma of these cells. Anti-OXT and auto-MS stained droplets in the cytoplasm of the fetal zone cells. Similar distinct staining patterns for AVP and OXT cells were obtained in human anencephalics. These observations show that the peptides are not derived from the fetal brain, but are rather produced in the fetal adrenal cortex. Future research will have to determine the physiological meaning of the presence of these peptides in the fetal adrenal, e.g., in their contribution to amniotic fluid peptides, their possible role in fetal stress, steroidogenesis etc. The presence of an alternative source of these peptides in the fetus makes it necessary, further-more, to reconsider their possible functions in the process of labour.In honour of Prof. P. van Duijn     

Localisation of oxytocin, vasopressin and parts of precursors in the human neonatal adrenal

Being a possible alternative source for the production of vasopressin (AVP) and oxytocin (OXT), a study was undertaken of the fetal adrenal. The concentrations of these peptides within the fetal adrenal turned out to be low, viz., approx. 1 pg/mg in the rat and within the pg/g range in the human. Immunocytochemistry was performed either on conventional autopsy material kept till 12 years in paraffin blocks, or on more recently obtained formalin or glutaraldehyde-paraformaldehyde fixed material. In both types of material staining was good. In order to localize AVP cells, anti-AVP, an antibody against its associated neurophysin (anti-NSN) or an antibody raised against the c-terminal glycopeptide part of the AVP precursor (anti-GP) was used. OXT cells were localized by means of anti-OXT or an auto-antibody of a multiple sclerosis patient (auto-MS) probably recognizing OXT-neurophysin. The antibodies were characterized on human and rat brain material. In the external zone of the definitive cortex, apart from parenchyma cells, anti-AVP, anti-NSN and anti-GP stained fibre-like structures running in the connective tissue septa and around parenchyma cells and the cytoplasma of these cells. Anti-OXT and auto-MS stained droplets in the cytoplasm of the fetal zone cells. Similar distinct staining patterns for AVP and OXT cells were obtained in human anencephalics. These observations show that the peptides are not derived from the fetal brain, but are rather produced in the fetal adrenal cortex.(ABSTRACT TRUNCATED AT 250 WORDS)     

Single-step isolation and resolution of pancreatic carboxypeptidases A and B.

Carboxypeptidases A and B have been isolated individually from aqueous extracts of mammalian pancreatic acetone powders by affinity chromatography on [N-(epsilon-aminocaproyl)-p-aminobenzyl]succinyl-Sepharose 4B (CABS-Sepharose). The affinity ligand was synthesized from DL-benzylsuccinic acid, purified, and characterized by UV absorption and NMR spectroscopy. Both enzymes from the various species were homogeneous by NaDodSO4-polyacrylamide gel electrophoresis and displayed high specific activities. No cross contamination of one enzyme species with the other was found. The ease of synthesis of the ligand from its commercially available precursor, its stability, and the mild elution conditions render CABS-Sepharose an excellent affinity support for the single-column isolation of both carboxypeptidases A and B. The procedures extend the utility of this resin previously demonstrated for carboxypeptidase A from human pancreatic juice [Peterson, L. M., Sokolovsky, M., & Vallee, B. L. (1976) Biochemistry, 15, 2501]. The use of CABS-Sepharose as a general affinity matrix for the isolation of metallocarboxypeptidases is suggested.     

Exposure to chronic isolation modulates receptors mRNAs for oxytocin and vasopressin in the hypothalamus and heart

The goal of our study was to explore the effect of social isolation stress of varying durations on the plasma oxytocin (OT), messenger ribonucleic acid (mRNA) for oxytocin receptor (OTR), plasma arginine vasopressin (AVP) and mRNA for V_1a receptor of AVP (V_1aR) expression in the hypothalamus and heart of socially monogamous female and male prairie voles (Microtus ochrogaster). Continuous isolation for 4 weeks (chronic isolation) increased plasma OT level in females, but not in males. One hour of isolation every day for 4 weeks (repeated isolation) was followed by a significant increase in plasma AVP level. Chronic isolation, but not repeated isolation, significantly decreased OTR mRNA in the hypothalamus and heart in both sexes. Chronic isolation significantly decreased cardiac V_1aR mRNA, but no effect on hypothalamic V_1aR mRNA expression. We did not find a gender difference within repeated social isolation groups. The results of the present study reveal that although chronic social isolation can down-regulate gene expression for the OTR in both sexes, the release of the OT peptide was increased after chronic isolation only in females, possibly somewhat protecting females from the negative consequences of isolation. In both sexes repeated, but not chronic, isolation increased plasma AVP, which could be permissive for mobilization and thus adaptive in response to a repeated stressor. The differential effects of isolation on OT and AVP systems may help in understanding mechanisms through social interactions can be protective against emotional and cardiovascular disorders.     

Effect of intraventricular oxytocin and vasopressin on self-stimulation in rats.

Oxytocin (500 mu u) and vasopressin (50 mu u) were injected into the lateral ventricle and its effect on hypothalamic self-stimulation has been studied. Oxytocin increased, while vasopressin decreased the self-stimulation rate tested 10-20 min following application. The hypothalamic and mesencephalic serotonin content decreased slightly while plasma corticosterone content did not change 20 min after oxytocin and vasopressin administration compared to the injected control animals. The data suggest that vasopressin and oxytocin have an opposite effect on self-stimulation and this action is not mediated through the brain serotoninergic or pituitary-adrenocortical axis.     

Reaction of tetranitromethane with lutropin, oxytocin, and vasopressin.

Tetranitromethane reaction with intact ovine lutropin and its isolated subunits was studied using spectrophotometric measurements, amino acid analysis, and isolation of tyrosyl peptides. Tyrosyl residues in the beta subunit (beta37, beta59) did not react with tetranitromethane in the intact hormone, but were nitrated in the isolated subunit. The sequence and extent of reaction of tetranitromethane with the tyrosyl residues in the alpha subunit was alpha21 = alpha92 = alpha93 (in intact hormone or isolated subunit) greater than alpha 41 (reacted in isolated subunit only) greater than alpha 30 (reacted in isolated subunit in 8 M urea only). Polymerization was observed as a side reaction in agreement with previous studies. The degree of polymerization appeared to be related to both primary sequence and tertiary structure, and for lutropin had the relation: alpha subunit (93% polymerized) greater than intact hormone greater than beta subunit (less than 40%). Polymerization observed with vasopressin was significantly greater than with oxytocin; for these peptides the tyrosine residues in the monomeric product were converted to 3-nitrotyrosine. Neither 3-nitrotyrosine nor tyrosine was detected in the polymerized by-products. In the tetranitromethane reaction with intact ovine lutropin, other reaction products charcterized by absorption spectra were found. Peptides isolated from these products lacked the characteristic 428 nm abosrption maxima of 3-nitrotyrosyl peptides and showed instead absorption in the 310 to 350 nm region. Similar products from tetranitromethane reactions with di- and tripeptides containing tyrosine have been observed previously (Boyd, N.D., and Smith, D.B. (1971) Can. J. Biochem, 49, 154-161), but they have not been studied in proteins. A possible relationship to the polymerization side reaction is suggested.     

Chromosomal and comparative mapping of rat oxytocin, oxytocin receptor and vasopressin genes

Oxytocin and its receptor are potentially important for cardiovascular functions. In the present paper, we report their chromosome locations in the rat and their comparative mapping with the mouse and human. They are located in chromosome regions previously known to contain quantitative trait loci for blood pressure in various genetic crosses. Thus, they have become valid candidate genes for genetic hypertension.     

催产素和加压素与应激的关系

催产素和加压素是由下丘脑视上核和室旁核大细胞性神经内分泌细胞合成和分泌的一种神经垂体激素。各种应刺激都可以引起催产素和加压素神经元的活动。目前应激后引起的这类神经活动的变化与人类的某结疾病的病理生理相关联正在引起人们的关注。西文总结了近几年在这方面的研究进展。主要内容包括：（1）催产素和加压素神经元在应激中的反应;（2）在大细胞性催产素和加压素神经元的应激反应相关联的神经传递物质;（3）与应激相关联的精神疾病的关系。     

Distribution of vasopressin and oxytocin in rat brain

Arginine-vasopressin and oxytocin in various portions of rat brain were determined by radioimmunoassays. The hormones were extracted from tissue samples into 0.1 N HCl and then purified partially with acetone-petroleum ether extraction. The non-equilibration method was used for the assays. In this method recovery rates of arginine-vasopressin and oxytocin were 73.0 +/- 4.4% and 75.0 +/- 3.8%, respectively. Sensitivities of the assays were 1 pg of arginine-vasopressin and 0.75 pg (0.3 microU) of oxytocin per assay tube. The higher concentrations of arginine-vasopressin and oxytocin were confirmed in the hypothalamo-neurohypophyseal system, where these hormones are synthesized, transported and stored. Relatively high concentrations of these hormones, especially oxytocin, were detected in spinal cord. Amygdala, hippocampus, limbic forebrain and pineal body contained a certain amount of arginine-vasopressin (2-20 pg/mg protein). Oxytocin (1-7 pg/mg protein) was also detected in amygdala, pons and medulla oblongata, pineal body and midbrain. The low concentrations of these hormones were also found in cerebral cortex and cerebellum.     

Functional architecture of vasopressin/oxytocin receptors

Three-dimensional models of G protein-coupled receptors (GPCR) have been defined using most experimental data available and protein modeling techniques. The endogenous ligand binding sites have been qualitatively described and putative receptor activation mechanisms have been proposed. The model has been recently refined to take into account recent crystallographic data. Most experimental results published are in excellent qualitative agreement with the initial model. We have undertaken to study more systematically by site directed mutagenesis the vasopressin/oxytocin receptor binding domain as a prototype of neuropeptide receptors. The experimental results are in very good agreement with the models. The residues responsible for the neuropeptide binding have been identified and confirm the predicted localization of the neuromediator in the transmembrane domain of the receptors. The side chain of the 8th residue of vasopressin interacts with a non-conserved receptor residue located in the first extracellular loop. As predicted from the model, this interaction is completely responsible for the selectivity of the ligand-receptor interaction. Finally, aromatic residues which allow the modulation of the efficacy of agonists have been identified.     

Human neuroimaging of oxytocin and vasopressin in social cognition

The neuropeptides oxytocin and vasopressin have increasingly been identified as modulators of human social behaviors and associated with neuropsychiatric disorders characterized by social dysfunction, such as autism. Identifying the human brain regions that are impacted by oxytocin and vasopressin in a social context is essential to fully characterize the role of oxytocin and vasopressin in complex human social cognition. Advances in human non-invasive neuroimaging techniques and genetics have enabled scientists to begin to elucidate the neurobiological basis of the influence of oxytocin and vasopressin on human social behaviors. Here we review the findings to-date from investigations of the acute and chronic effects of oxytocin and vasopressin on neural activity underlying social cognitive processes using "pharmacological fMRI" and "imaging genetics", respectively. This article is part of a Special Issue entitled Oxytocin, Vasopressin, and Social Behavior.