bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of Cellular Interactions on Calcium Dynamics in Prolactin-Secreting Cells*
Authors: Abraham Elizabeth J. , Villalobos Carlos , Frawley L. Stephen 
Published: January 2014

AbstractSignals derived from other pituitary cells can have a dramatic effect on PRL gene expression and secretion by mammotropes. However, the intracellular mechanisms by which these effects are manifested on the target cell remain unexplored. Inasmuch as calcium is a key modulator of both gene expression and hormone export in mammotropes, we evaluated the effects of cell to cell contact vs. specific cellular interactions on calcium dynamics within these cells. This was accomplished by digital-imaging fluorescence microscopy of fura-2 in pituitary cells that were isolated in culture (singles) or adjoining one other cell (doublets). After calcium imaging, we then subjected cells to immunocytochemistry for PRL. Doublets were further categorized into mammotropes attached to another mammotrope (M-M) or to a nonmammotrope (M-nonM). We then calculated and compared Mean[ Ca2+]i values as well as Oscillation Indices (which reflect the oscillatory behavior of cells) in singles and doublets and found that they were not different (P&gt; 0.05). However, the phenotype of the adjoining cell had a profound influence on both of these calcium parameters, such that the presence of one mammotrope could consistently decrease (P &lt; 0.05) the Mean [Ca2+]i value (39.17 ± 3.83 vs. 56.24 ± 5.56 in M-nonM) and Oscillation Index (10.19 ± 1.76 vs. 21.21 ± 3.73 in M-nonM) of its neighboring counterpart. A more detailed analysis of oscillatory patterns in these cells revealed that nonoscillators were more abundant in M-M (23%) than in M-nonM (12%) doublets. Taken together, our results indicate that PRL-secreting cells convey a signal that dampens the oscillatory behavior of neighboring mammotropes. Thus, it appears that it is the phenotype rather than the physical presence of a neighbor that controls intercellular regulation of calcium dynamics among mammotropes.
Corticosterone Can Facilitate as Well as Inhibit Corticotropin-Releasing Hormone Gene Expression in the Rat Hypothalamic Paraventricular Nucleus*
Authors: Tanimura Susan M. , Watts Alan G. 
Published: January 2014

Abstract
We have used in situ hybridization to investigate how basal levels of circulating corticosterone modulate CRH gene transcription in the neuroendocrine parvicellular part of the hypothalamic paraventricular nucleus (PVHmpd) during sustained hypovolemia. In the absence of the stressor, the accumulation rate of the CRH primary transcript exhibited a dose dependency on low maintained levels of plasma corticosterone similar to that previously reported for the mature messenger RNA (mRNA); levels declined as plasma corticosterone increased. In response to hypovolemia, the absence of corticosterone compromised CRH gene transcription mechanisms to mount the activated response seen in intact animals. However, adrenalectomized rats with low doses of corticosterone (insufficient to normalize thymus weights) showed an augmented mRNA response compared with that in intact animals. When replaced corticosterone normalized thymus weights, the magnitude of the mRNA response was reduced to that seen in intact animals. In contrast to CRH gene regulation, PVHmpd proenkephalin mRNA levels were unaffected by corticosterone concentrations. These results suggest that corticosterone affects CRH gene transcription in the PVHmpd using two mechanisms: first, inhibition, which probably uses type II glucocorticoid receptor-dependent mechanisms and contributes to classic negative feedback; and second, facilitation, which is seen at low plasma concentrations and maintains gene transcription in the presence of sustained stress, possibly using type I mechanisms. This suggests that one reason why adrenal insufficiency severely compromises survival of sustained stress is that CRH gene transcription cannot be maintained without previous exposure to low levels of plasma corticosterone.
Intracisternal Antisense Oligodeoxynucleotides to the Thyrotropin-Releasing Hormone Receptor Blocked Vagal-Dependent Stimulation of Gastric Emptying Induced by Acute Cold in Rats*
Authors: Martinez V. , Wu S. V. , Taché Y. 
Published: January 2014

Abstract
Cold exposure increases TRH gene expression in hypothalamic and raphe nuclei and results in a vagal activation of gastric function. We investigated the role of medullary TRH receptors in cold (4–6 C, 90 min)-induced stimulation of gastric motor function in fasted conscious rats using intracisternal injections of TRH receptor (TRHr) antisense oligodeoxynucleotides (100 μg twice, −48 and −24 h). The gastric emptying of a methyl-cellulose solution was assessed by the phenol red method. TRH (0.1 μg) or the somatostatin subtype 5-preferring analog, BIM-23052 (1 μg), injected intracisternally increased basal gastric emptying by 34% and 47%, respectively. TRHr antisense, which had no effect on basal emptying, blocked TRH action but did not influence that of BIM-23052. Cold exposure increased gastric emptying by 64%, and the response was inhibited by vagotomy, atropine (0.1 mg/kg, ip), and TRHr antisense (intracisternally). Saline or mismatched oligodeoxynucleotides, injected intracisternally under similar conditions, did not alter the enhanced gastric emptying induced by cold or intracisternal injection of TRH or BIM-23052. These results indicate that TRH receptor activation in the brain stem mediates acute cold-induced vagal cholinergic stimulation of gastric transit, and that medullary TRH may play a role in the autonomic visceral responses to acute cold.
Fasting Regulates Hypothalamic Neuropeptide Y, Agouti-Related Peptide, and Proopiomelanocortin in Diabetic Mice Independent of Changes in Leptin or Insulin1
Authors: Mizuno Tooru M. , Makimura Hideo , Silverstein Jeffrey , Roberts James L. , Lopingco Tina , Mobbs Charles V. 
Published: January 2014

Abstract
Fasting increases hypothalamic neuropeptide Y (NPY) and agouti-related peptide (AGRP) messenger RNA (mRNA) and reduces hypothalamic POMC mRNA, and is also characterized by a reduction in plasma leptin, insulin, and glucose, each of which has been implicated in the regulation of hypothalamic gene expression. To further evaluate the roles of leptin, insulin, and glucose in mediating effects of fasting, we examined hypothalamic gene expression in nondiabetic and streptozotocin (STZ)-induced diabetic mice both under ad lib fed and 48-h fasted conditions. In both diabetic and nondiabetic mice, fasting stimulated hypothalamic NPY and AGRP mRNA and inhibited hypothalamic POMC mRNA and adipose leptin mRNA. However, in diabetic mice fasting had no effect on plasma leptin and insulin while decreasing plasma glucose, whereas in nondiabetic mice fasting decreased plasma leptin, insulin, and glucose. Furthermore, in nondiabetic fasted mice, NPY and AGRP mRNA were higher, and POMC mRNA and plasma glucose were lower, than in diabetic ad lib fed mice, even though insulin and leptin were similar in these two groups. These data are consistent with the hypothesis that although leptin and insulin regulate hypothalamic gene expression, glucose or other factors may have independent effects on hypothalamic and adipose gene expression under conditions of low insulin and leptin.
Posttranslational Processing of Progrowth Hormone-Releasing Hormone1
Authors: Nillni Eduardo A. , Steinmetz Rosemary , Pescovitz Ora Hirsch 
Published: January 2014

AbstractThe prepro-GH-releasing hormone (prepro-GHRH; 12.3 kDa) precursor, like other neuropeptide precursors, undergoes proteolytic cleavage to give rise to mature GHRH, which is the primary stimulatory regulator of pituitary GH secretion. In this study we present the first model of in vitro pro-GHRH processing. Using pulse-chase analysis, we demonstrate that at least five peptide forms in addition to GHRH are produced. The pro-GHRH (after removal of its signal peptide, 10.5 kDa) is first processed to an 8.8-kDa intermediate form that is cleaved to yield two products: the 5.2-kDa GHRH and GHRH-related peptide (GHRH-RP; 3.6 kDa). GHRH-RP is a recently described peptide derived from proteolytic processing of pro-GHRH that activates stem cell factor, a factor known to be essential for hemopoiesis, spermatogenesis, and melanocyte function. Further cleavage results in a 3.5-kDa GHRH and a 2.2-kDa product of GHRH-RP.Like GHRH, there is GHRH-RP immunostaining in hypothalamic neurons in the median eminence as detected by immunohistochemistry and immunoelectron microscopy. Based on deduced amino acid sequences of the pro-GHRH processing products, several peptides were synthesized and tested for their ability to stimulate the cAMP second messenger system. GHRH, GHRH-RP, and one of these peptides[ prepro-GHRH-(75–92)-NH2] all significantly stimulated the PKA pathway. This work delineates a new model of pro-GHRH processing and demonstrates that novel peptides derived from this processing may have biological action.
Interaction between Hypothalamic Dopaminergic and Opioidergic Systems in the Photoperiodic Regulation of Pulsatile Luteinizing Hormone Secretion in Sheep
Authors: Tortonese Domingo J. 
Published: January 2014

Abstract
Previous studies in sheep have shown that whereas the inhibitory effects of dopamine (DA) systems on GnRH/gonadotrophin secretion are readily detectable during the sexually inactive phase under long days (LD), the suppressive effects of endogenous opioid peptide (EOP) systems are most evident during the sexually active phase under short days (SD). The hypothesis proposed in this study is that inhibitory DA pathways interact with EOP neurons to regulate GnRH/gonadotropin secretion in sheep and that photoperiod modulates this interaction to relay its effect on the seasonal reproductive cycle. To test this hypothesis, the effects of a DA agonist (bromocriptine) or of a DA antagonist (sulpiride) on the pulsatile LH response to an opioid antagonist (naloxone) were evaluated in sexually active Soay rams exposed to SD, and then reassessed when sexually inactive under LD. The experimental design comprised six treatments: 1) control (vehicle); 2) bromocriptine; 3) sulpiride; 4) naloxone; 5) pretreatment with bromocriptine followed by naloxone; 6) pretreatment with sulpiride followed by naloxone. Under SD, when DA pathways are thought to be quiescent and EOP systems active, bromocriptine suppressed pulsatile LH secretion (P &lt; 0.01), whereas sulpiride had no effect. Under this photoperiod, naloxone induced a conspicuous stimulation of episodic LH release (P &lt; 0.01). This effect was prevented by pretreatment with bromocriptine (P &lt; 0.01), but was not affected by pretreatment with sulpiride. Conversely, under LD, when the activity of DA pathways is thought to be increased and that of EOP systems reduced, bromocriptine was without effect, whereas sulpiride evoked a mild increase in LH pulse frequency (P &lt; 0.05). Under this photoperiod, naloxone induced a smaller stimulation than under SD. This effect was again blocked by pretreatment with bromocriptine but, in contrast to SD, markedly enhanced by pretreatment with sulpiride (P &lt; 0.01). Particularly relevant was that the DA agonist blocked the stimulatory effects of the EOP antagonist under SD, and that the DA antagonist enhanced the effects of the EOP antagonist only under LD. These results are consistent with the hypothesis proposing that, in sheep, DA pathways have a predominant inhibitory effect on both GnRH and EOP neurons, and that changes in day length modulate the interplay between DA and EOP systems as part of the mechanisms involved in the photoperiodic control of the seasonal reproductive cycle.
RORα Augments Thyroid Hormone Receptor-Mediated Transcriptional Activation*
Authors: Koibuchi Noriyuki , Liu Ying , Fukuda Harumi , Takeshita Akira , Yen Paul M. , Chin William W. 
Published: January 2014

AbstractThis study is designed to clarify the role of an orphan nuclear hormone receptor, RORα, on thyroid hormone (TH) receptor (TR)-mediated transcription on a TH-response element (TRE). A transient transfection study using various TREs [i.e., F2 (chick lysozyme TRE), DR4 (direct repeat), and palindrome TRE] and TR and RORα1 was performed. When RORα1 and TR were cotransfected into CV1 cells, RORα1 enhanced the transactivation by liganded-TR on all TREs tested without an effect on basal repression by unliganded TR. By electrophoretic mobility shift assay, on the other hand, although RORα bound to all TREs tested as a monomer, no (or weak) TR and RORα1 heterodimer formation was observed on various TREs except when a putative ROR-response element was present. The transactivation by RORα1 on a ROR-response element, which does not contain a TRE, was not enhanced by TR. The effect of RORα1 on the TREs is unique, because, whereas other nuclear hormone receptors (such as vitamin D receptor) may competitively bind to TRE to exert dominant negative function, RORα1 augmented TR action. These results indicate that RORα1 may modify the effect of liganded TR on TH-responsive genes. Because TR and RORα are coexpressed in cerebellar Purkinje cells, and perinatal hypothyroid animals and RORα-disrupted animals show similar abnormalities of this cell type, cross-talk between these two receptors may play a critical role in Purkinje cell differentiation.
Uterine Natural Killer Cells Are Targets for a Trophoblast Cell-Specific Cytokine, Prolactin-Like Protein A*
Authors: Müller Heiner , Liu Bing , Croy B. Anne , Head Judith R. , Hunt Joan S. , Dai Guoli , Soares Michael J. 
Published: January 2014

AbstractPRL-like protein A (PLP-A) is a member of the PRL family expressed in trophoblast cells coincident with establishment of the chorioallantoic placenta. The purpose of this investigation was to identify targets for PLP-A. Using an alkaline phosphatase-tagging strategy, we show that PLP-A specifically interacts with a population of natural killer (NK) lymphocytes within the mesometrial compartment of decidua from pregnant and pseudopregnant rats. These observations are supported by the codistribution of PLP-A targets with cells expressing the rat NK cell surface marker, gp42, the absence of PLP-A binding in conceptuses from NK cell-deficient tgε26 mice, and the specific interaction of PLP-A with a rat NK cell line, RNK-16. We have further demonstrated that PLP-A effectively suppresses RNK-16 cell cytolytic activities. Our results provide evidence for a new paradigm of embryonic-maternal communication involving a PLP-A signaling pathway between trophoblast cells and uterine NK lymphocytes.
Expression, Processing, and Secretion of the Neuroendocrine VGF Peptides by INS-1 Cells*
Authors: Possenti Roberta , Rinaldi Anna Maria , Ferri Gian-Luca , Borboni Patrizia , Trani Eugenia , Levi Andrea 
Published: January 2014

AbstractThe neurotropin-inducible gene vgf is expressed in neuronal and endocrine tissues. It encodes a secretory protein that is proteolytically processed in neuronal cells to low molecular mass polypeptides. In the present report, we show that vgf is expressed in different insulinoma cell lines and in normal rat pancreatic islets. In the insulinoma-derived β-cell line INS-1, vgf messenger RNA was transcriptionally up-regulated by increased levels of intracellular cAMP, but not by the addition of glucose (20 mm) or phorbol 12-myristate 13-acetate (100 nm). Furthermore, nerve growth factor failed to stimulate vgf gene expression. In INS-1 cells, the VGF protein was shown to be processed in a post endoplasmic reticulum compartment to produce a peptide profile similar to that seen in neurons. The release of such VGF peptides occurred at a low rate in the absence of secretory stimuli (&lt;2%/h). A 3-fold increase in the rate of release was seen after the addition of glucose (15 mm), a 4-fold increase was seen after (Bu)2cAMP (1 mm), and a 6-fold increase was seen after phorbol 12-myristate 13-acetate (100 nm). These results indicated that insulin-containing cells produce VGF-derived peptides that are released via a regulated pathway in response to insulin secretagogues.
The Human Growth Hormone Antagonist B2036 Does Not Interact with the Prolactin Receptor
Authors: Goffin Vincent , Bernichtein Sophie , Carrière Océane , Bennett William F. , Kopchick John J. , Kelly Paul A. 
Published: January 2014

Abstract
The human growth hormone (hGH) antagonist B2036 combines a single amino acid substitution impairing receptor binding site 2 (G120K) with eight additional amino acid substitutions that improve binding site 1 affinity. This hGH antagonist is being tested for treating pathologies linked to excess hGH levels. B2036-PEG is a polyethylene glycol (PEG) conjugated form of B2036 that has an increased half-life due to reduced renal clearance. It is currently in phase III trials for acromegaly. Human GH is also able to bind to the receptor of prolactin (PRLR). Since activation of PRLR can promote an array of pathological states (reproduction disorders, breast cancer), the ability of B2036-PEG to interact with the PRLR had to be determined. In this study, we compared four hGH antagonists (G120K, G120K-PEG, B2036 and B2036-PEG) in three bioassays: proliferation of rat Nb2 cells, binding to the human PRLR and activation of human PRLR-mediated signaling in a cell line stably expressing this receptor and a luciferase reporter gene. Agonistic and antagonistic properties were characterized. Our data show that B2036-PEG does not bind, activate or antagonize PRLRs, either from rat or human origin. These observations further demonstrate that the eight amino acid substitutions within binding site 1 provide binding specificity directed towards the human GH receptor.
Glucosamine Regulation of Glucose Metabolism in Cultured Human Skeletal Muscle Cells: Divergent Effects on Glucose Transport/Phosphorylation and Glycogen Synthase in Non-Diabetic and Type 2 Diabetic Subjects1
Authors: Ciaraldi Theodore P. , Carter Leslie , Nikoulina Svetlana , Mudaliar Sunder , McClain Donald A. , Henry Robert R. 
Published: January 2014

AbstractChronic exposure (48 h) to glucosamine resulted in a dose-dependent reduction of basal and insulin-stimulated glucose uptake activities in human skeletal muscle cell cultures from nondiabetic and type 2 diabetic subjects. Insulin responsiveness of uptake was also reduced. There was no change in total membrane expression of either GLUT1, GLUT3, or GLUT4 proteins. While glucosamine treatment had no significant effects on hexokinase activity measured in cell extracts, glucose phosphorylation in intact cells was impaired after treatment. Under conditions where glucose transport and phosphorylation were down regulated, the fractional velocity (FV) of glycogen synthase was increased by glucosamine treatment. Neither the total activity nor protein expression of glycogen synthase were influenced by glucosamine treatment. The stimulation of glycogen synthase by glucosamine was not due totally to soluble mediators. Reflective of the effects on transport/phosphorylation, total glycogen content and net glycogen synthesis were reduced after glucosamine treatment. These effects were similar in nondiabetic and type 2 cells. In summary: 1) Chronic treatment with glucosamine reduces glucose transport/phosphorylation and storage into glycogen in skeletal muscle cells in culture and impairs insulin responsiveness as well. 2) Down-regulation of glucose transport/phosphorylation occurs at a posttranslational level of GLUTs. 3) Glycogen synthase activity increases with glucosamine treatment. 4) Nondiabetic and type 2 muscle cells display equal sensitivity and responsiveness to glucosamine. Increased exposure of skeletal muscle to glucosamine, a substrate/precursor of the hexosamine pathway, alters intracellular glucose metabolism at multiple sites and can contribute to insulin resistance in this tissue.
Pituitary Hormone Gene Expression and Secretion in Human Growth Hormone-Releasing Hormone Transgenic Mice: Focus on Lactotroph Function1
Authors: Moore Joseph P. , Cai Aihua , Hostettler Mary Ellen , Arbogast Lydia A. , Voogt James L. , Hyde James F. 
Published: January 2014

Abstract
The human GH-releasing hormone (hGHRH) transgenic mouse has a hyperplastic anterior pituitary gland that eventually develops into an adenoma. We showed previously that the number of lactotrophs in the male hGHRH transgenic mouse is increased 2-fold, yet there is no concomitant increase in plasma levels of PRL. To further elucidate underlying changes in lactotroph function in the hGHRH transgenic mouse, the objectives of this study were to 1) examine the relative differences in PRL gene expression in transgenic mice and their siblings, 2) quantify PRL secretion at the level of the individual cell, 3) determine whether tyrosine hydroxylase gene expression and/or activity are altered in the hypothalamus of transgenic mice, and 4) assess dopamine receptor gene expression and functional sensitivity in lactotrophs of transgenic mice. Total PRL messenger RNA (mRNA) levels were increased nearly 5-fold in the hGHRH transgenic mouse, whereas the concentrations of PRL mRNA (PRL mRNA per μg total RNA) were unchanged. In contrast, total PRL contents were unchanged, whereas the concentrations of PRL (micrograms of PRL per mg total protein) were decreased 3-fold. Hypothalamic tyrosine hydroxylase steady state mRNA levels were not altered in the hGHRH transgenic mice, but hypothalamic tyrosine hydroxylase activity was increased 2-fold in transgenic mice. Dopamine D2 receptor mRNA concentrations in the anterior pituitary were increased 2.5-fold in hGHRH transgenic mice, and total pituitary D2 receptor mRNA levels were increased nearly 10-fold. Furthermore, the basal secretory capacity of lactotrophs from transgenic mice was increased significantly at the level of the single cell, and dopamine inhibited the secretion of PRL to a greater extent in hGHRH transgenic mice. Thus, although the total number of lactotrophs is increased 2-fold in hGHRH transgenic mice, the present data are consistent with the hypothesis that increased hypothalamic dopamine synthesis and release coupled with an increase in D2 dopamine receptor gene expression and functional sensitivity in the pituitary result in normal plasma levels of PRL.