scholarly journals Characterization of Neuropeptide B (NPB), Neuropeptide W (NPW), and Their Receptors in Chickens: Evidence for NPW Being a Novel Inhibitor of Pituitary GH and Prolactin Secretion

Endocrinology ◽  
2016 ◽  
Vol 157 (9) ◽  
pp. 3562-3576 ◽  
Author(s):  
Guixian Bu ◽  
Dongliang Lin ◽  
Lin Cui ◽  
Long Huang ◽  
Can Lv ◽  
...  
Keyword(s):  
Amino Acids ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Prolactin Secretion ◽  
Luciferase Reporter ◽  
Intracellular Camp ◽  
Pituitary Cells ◽  
Reporter System ◽  
Ovary Cells ◽  
The Central Nervous System

The 2 structurally and functionally related peptides, neuropeptide B (NPB) and neuropeptide W (NPW), together with their receptor(s) (NPBWR1/NPBWR2) constitute the NPB/NPW system, which acts mainly on the central nervous system to regulate many physiological processes in mammals. However, little is known about this NPB/NPW system in nonmammalian vertebrates. In this study, the functionality and expression of this NPB/NPW system and its actions on the pituitary were investigated in chickens. The results showed that: 1) chicken NPB/NPW system comprises an NPB peptide of 28 amino acids (cNPB28), an NPW peptide of 23 or 30 amino acids (cNPW23/cNPW30), and their 2 receptors (cNPBWR1 and cNPBWR2), which are highly homologous to their human counterparts. 2) Using a pGL3-CRE-luciferase reporter system, we demonstrated that cNPBWR2 expressed in Chinese hamster ovary cells can be potently activated by cNPW23 (not cNPB28), and its activation inhibits the intracellular cAMP signaling pathway, whereas cNPBWR1 shows no response to peptide treatment, suggesting a crucial role of cNPBWR2 in mediating cNPW/cNPB actions. 3) Quantitative real-time PCR revealed that cNPW and cNPB are widely expressed in chicken tissues, including hypothalamus, whereas cNPBWR1 and cNPBWR2 are mainly expressed in brain or pituitary. 4) In accordance with abundant cNPBWR2 expression in pituitary, cNPW23 could dose dependently inhibit GH and prolactin secretion induced by GHRH and vasoactive intestinal polypeptide, respectively, in cultured chick pituitary cells, as monitored by Western blotting. Collectively, our data reveal a functional NPB/NPW system in birds and offer the first proof that NPW can act directly on pituitary to inhibit GH/prolactin secretion in vertebrates.

Metabolism of peptide amino acids by Chinese hamster ovary cells grown in a complex medium

1999 ◽  
Vol 62 (3) ◽  
pp. 324-335 ◽  
Author(s):  
Gregg B. Nyberg ◽  
R. Robert Balcarcel ◽  
Brian D. Follstad ◽  
Gregory Stephanopoulos ◽  
Daniel I. C. Wang
Keyword(s):  
Amino Acids ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Complex Medium ◽  
Ovary Cells

scholarly journals Truncations of the C-terminal cytoplasmic domain of MG160, a medial Golgi sialoglycoprotein, result in its partial transport to the plasma membrane and filopodia

1998 ◽  
Vol 111 (2) ◽  
pp. 249-260 ◽  
Author(s):  
J.O. Gonatas ◽  
Y.J. Chen ◽  
A. Stieber ◽  
Z. Mourelatos ◽  
N.K. Gonatas
Keyword(s):  
Amino Acids ◽  
Plasma Membrane ◽  
Golgi Apparatus ◽  
Cell Surface ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Growth Factor Receptor ◽  
Cytoplasmic Domain ◽  
Type I ◽  
Ovary Cells

MG160, a type I cysteine-rich membrane sialoglycoprotein residing in the medial cisternae of the rat Golgi apparatus, is highly homologous to CFR, a fibroblast growth factor receptor, and ESL-1, an E-selectin ligand located at the cell surface of mouse myeloid cells and recently detected in the Golgi apparatus as well. The mechanism for the transport of MG160 from the Golgi apparatus to the cell surface is unknown. In this study we found that differential processing of the carboxy-terminal cytoplasmic domain (CD), consisting of amino acids Arg1159 Ile Thr Lys Arg Val Thr Arg Glu Leu Lys Asp Arg1171, resulted in the partial transport of the protein to the plasma membrane and filopodia. In Chinese hamster ovary cells (CHO), stably transfected with the entire cDNA encoding MG160, the protein was localized in the Golgi apparatus. However, when the terminal Arg1171 or up to nine distal amino acids were deleted, the protein was distributed to the plasma membrane and filopodia as well as the Golgi apparatus. This report shows that the CD of an endogenous type I Golgi protein is important for its efficient retention and identifies a unique residue preference in this process. Cleavage within the CD of MG160 may constitute a regulatory mechanism for the partial export of the protein from the Golgi apparatus to the plasma membrane and filopodia.

scholarly journals Insulin-Like Growth Factor-I Provokes Functional Antagonism and Internalization of β1-Adrenergic Receptors

Endocrinology ◽  
2007 ◽  
Vol 148 (6) ◽  
pp. 2653-2662 ◽  
Author(s):  
Shai Gavi ◽  
Dezhong Yin ◽  
Elena Shumay ◽  
Hsien-yu Wang ◽  
Craig C. Malbon
Keyword(s):  
Cardiac Myocytes ◽  
Tyrosine Kinases ◽  
Adrenergic Receptors ◽  
Adrenergic Receptor ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Intracellular Camp ◽  
Ovary Cells ◽  
Igf I ◽  
Protein Kinase Akt

Hormones that activate receptor tyrosine kinases have been shown to regulate G protein-coupled receptors, and herein we investigate the ability of IGF-I to regulate the β1-adrenergic receptor. Treating Chinese hamster ovary cells in culture with IGF-I is shown to functionally antagonize the ability of expressed β1-adrenergic receptors to accumulate intracellular cAMP in response to stimulation by the β-adrenergic agonist Iso. The attenuation of β1-adrenergic action was accompanied by internalization of β1-adrenergic receptors in response to IGF-I. Inhibiting either phosphatidylinositol 3-kinase or the serine/threonine protein kinase Akt blocks the ability of IGF-I to antagonize and to internalize β1-adrenergic receptors. Mutation of one potential Akt substrate site Ser412Ala, but not another Ser312Ala, of the β1-adrenergic receptor abolishes the ability of IGF-I to functionally antagonize and to sequester the β1-adrenergic receptor. We also tested the ability of IGF-I to regulate β1-adrenergic receptors and their signaling in adult canine cardiac myocytes. IGF-I attenuates the ability of β1-adrenergic receptors to accumulate intracellular cAMP in response to Iso and promotes internalization of β1-adrenergic receptors in these cardiac myocytes.

scholarly journals The A, ASC, and L systems for the transport of amino acids in Chinese hamster ovary cells (CHO-K1).

1981 ◽  
Vol 256 (20) ◽  
pp. 10259-10266
Author(s):  
R. Bass ◽  
H.B. Hedegaard ◽  
L. Dillehay ◽  
J. Moffett ◽  
E. Englesberg
Keyword(s):  
Amino Acids ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
L Systems

scholarly journals Recessive constitutive mutant Chinese hamster ovary cells (CHO-K1) with an altered A system for amino acid transport and the mechanism of gene regulation of the A system.

1984 ◽  
Vol 4 (4) ◽  
pp. 799-808 ◽  
Author(s):  
J Moffett ◽  
E Englesberg
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Chinese Hamster Ovary ◽  
Amino Acid Transport ◽  
Actinomycin D ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Acid Transport ◽  
Ovary Cells ◽  
Proline Transport

Chinese hamster ovary cells (CHO-K1) starved for 24 h for amino acids show a severalfold increase in velocity of proline transport through the A system (Vmax is five times that of unstarved cells). This increase is inhibited by cycloheximide, actinomycin D, N-methyl-alpha-amino isobutyric acid (MeAIB, a non-metabolizable specific A system amino acid analog), and by other amino acids that are generally transported by the A system. However, transport by the A system is not a prerequisite for this repression, and all compounds that have affinity for the A system do not necessarily act as "co-repressors." The addition of proline, MeAIB, or other amino acids, as described above, to derepressed cells results in a rapid decrease in A system activity. As shown with proline and MeAIB, this decrease in activity is in part due to a rapid trans-inhibition and a slow, irreversible inactivation of the A system. Neither process is inhibited by cycloheximide or actinomycin D. Alanine antagonizes the growth of CHO-K1 pro cells by preventing proline transport, and alanine-resistant mutants (alar) have been isolated (Moffett et al., Somatic Cell Genet. 9:189-213, 1983). alar2 and alar4 are partial and full constitutive mutants for the A system and have two and six times the Vmax for proline uptake by the A system, respectively. The A system in alar4 is also immune to the co-repressor-induced inactivation. Both alar2 and alar4 phenotypes are recessive. Alar3 shows an increase in Vmax and Km for proline transport through the A system, and this phenotype is codominant. All three mutants have a pleiotropic effect, producing increases in activity of the ASC and P systems of amino acid transport. This increase is not due to an increase in the Na+ gradient. The ASC and P phenotypes behave similarly to the A system in hybrids. A model has been proposed incorporating these results.

Lamellipodin proline rich peptides associated with native plasma butyrylcholinesterase tetramers

2008 ◽  
Vol 411 (2) ◽  
pp. 425-432 ◽  
Author(s):  
He Li ◽  
Lawrence M. Schopfer ◽  
Patrick Masson ◽  
Oksana Lockridge
Keyword(s):  
Nervous System ◽  
Neuromuscular Junctions ◽  
Therapeutic Potential ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
Tetramerization Domain ◽  
The Central Nervous System ◽  
Tail Protein ◽  
Proline Rich Peptide

BChE (butyrylcholinesterase) protects the cholinergic nervous system from organophosphorus nerve agents by scavenging these toxins. Recombinant human BChE produced from transgenic goat to treat nerve agent intoxication is currently under development. The therapeutic potential of BChE relies on its ability to stay in the circulation for a prolonged period, which in turn depends on maintaining tetrameric quaternary configuration. Native human plasma BChE consists of 98% tetramers and has a half-life (t½) of 11–14 days. BChE in the neuromuscular junctions and the central nervous system is anchored to membranes through interactions with ColQ (AChE-associated collagen tail protein) and PRiMA (proline-rich membrane anchor) proteins containing proline-rich domains. BChE prepared in cell culture is primarily monomeric, unless expressed in the presence of proline-rich peptides. We hypothesized that a poly-proline peptide is an intrinsic component of soluble plasma BChE tetramers, just as it is for membrane-bound BChE. We found that a series of proline-rich peptides was released from denatured human and horse plasma BChE. Eight peptides, with masses from 2072 to 2878 Da, were purified by HPLC and sequenced by electrospray ionization tandem MS and Edman degradation. All peptides derived from the same proline-rich core sequence PSPPLPPPPPPPPPPPPPPPPPPPPLP (mass 2663 Da) but varied in length at their N- and C-termini. The source of these peptides was identified through database searching as RAPH1 [Ras-associated and PH domains (pleckstrin homology domains)-containing protein 1; lamellipodin, gi:82581557]. A proline-rich peptide of 17 amino acids derived from lamellipodin drove the assembly of human BChE secreted from CHO (Chinese-hamster ovary) cells into tetramers. We propose that the proline-rich peptides organize the 4 subunits of BChE into a 340 kDa tetramer, by interacting with the C-terminal BChE tetramerization domain.

scholarly journals Evidence for the existence of a local activin–follistatin negative feedback loop in the goldfish pituitary and its regulation by activin and gonadal steroids

2007 ◽  
Vol 195 (3) ◽  
pp. 373-384 ◽  
Author(s):  
Gheorghe F Y Cheng ◽  
Chi-Wai Yuen ◽  
Wei Ge
Keyword(s):  
Feedback Loop ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Pituitary Hormones ◽  
Gonadal Steroids ◽  
Pituitary Cells ◽  
Ovary Cells ◽  
Wide Range

Activin is an important regulator of gonadotropin expression and production in the vertebrate pituitary, and its activity is fine-tuned by its binding protein follistatin. In the present study, a full-length cDNA for follistatin was cloned in the goldfish, which shows 74% amino acid sequence identity with that of mammals. Recombinant goldfish follistatin expressed in the Chinese hamster ovary cells significantly blocked activin-induced F5-5 cell differentiation. Goldfish follistatin is expressed in a wide range of tissues including the brain, pituitary, ovary, and testis. The expression of follistatin mRNA in the pituitary is regulated by both activin and gonadal steroids in vitro. Treatment with goldfish activin B for 48 h significantly up-regulated follistatin expression in cultured pituitary cells, suggesting a closed activin–follistatin feedback loop in the pituitary. In agreement with this, both human and goldfish follistatin down-regulated the expression of follistatin itself, probably due to the neutralization of endogenous activin. Examination of FSHβ and LHβ expression in the same samples supports the role of activin and follistatin in the differential regulation of FSH and LH as demonstrated previously. Since the expression level of activin βB in the pituitary is rather stable both in vitro and in vivo, it is conceivable that follistatin may play a pivotal regulatory role in the intra-pituitary activin system. Both estradiol and testosterone up-regulated follistatin expression in vitro, suggesting a mediating role for follistatin in steroid feedback on pituitary hormones. These results provide clues to the potential physiological roles of activin–follistatin system in the regulation of gonadotropin biosynthesis in teleosts.

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