A P2X7 receptor stimulates plasma membrane trafficking in the FRTL rat thyrocyte cell line

2004 ◽  
Vol 287 (4) ◽  
pp. C992-C1002 ◽  
Author(s):  
M. Y. Kochukov ◽  
A. K. Ritchie
Keyword(s):  
Plasma Membrane ◽  
Cell Line ◽  
Fluorescence Imaging ◽  
Receptor Activation ◽  
Disulfonic Acid ◽  
Receptor Subtypes ◽  
Patch Clamp Recording ◽  
Thyroid Cells ◽  
Rat Thyroid ◽  
Stimulation Of

Thyroid cells express a variety of P2Y and P2X purinergic receptor subtypes. G protein-coupled P2Y receptors influence a wide variety of thyrocyte-specific functions; however, functional P2X receptor-gated channels have not been observed. In this study, we used whole cell patch-clamp recording and fluorescence imaging of the plasma membrane marker FM1-43 to examine the effects of extracellular ATP on membrane permeability and trafficking in the Fisher rat thyroid cell line FRTL. We found a cation-selective current that was gated by ATP and 2′,3′- O-(4-benzoylbenzoyl)-ATP but not by UTP. The ATP-evoked currents were inhibited by pyridoxal phosphate 6-azophenyl-2′,4′-disulfonic acid, adenosine 5′-triphosphate-2′,3′-dialdehyde, 100 μM Zn2+, and 50 μM Cu2+. Fluorescence imaging revealed pronounced, temperature-sensitive stimulation of exocytosis and membrane internalization by ATP with the same pharmacological profile as observed for activation of current. The EC50 for ATP stimulation of internalization was 440 μM in saline containing 2 mM Ca2+ and 2 mM Mg2+, and 33 μM in low-Mg2+, nominally Ca2+-free saline. Overall, the results are most consistent with activation of a P2X7 receptor by ATP4−. However, low permeability to N-methyl-d-glucamine+ and the propidium cation YO-PRO-1 indicates absence of the cytolytic pore that often accompanies P2X7 receptor activation. ATP stimulation of internalization occurs in Na+-free, Ca2+-free, or low-Mg2+ saline and therefore does not depend on cation influx through the ATP-gated channel. We conclude that ATP activation of a P2X7 receptor stimulates membrane internalization in FRTL cells via a transduction pathway that does not depend on cation influx.

scholarly journals The sarcoplasmic–endoplasmic reticulum Ca2+ ATPase 2b regulates the Ca2+ transients elicited by P2Y2 activation in PC Cl3 thyroid cells

2006 ◽  
Vol 190 (3) ◽  
pp. 641-649 ◽  
Author(s):  
Luca Ulianich ◽  
Maria Giovanna Elia ◽  
Antonella Sonia Treglia ◽  
Antonella Muscella ◽  
Bruno Di Jeso ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Thyroid Cell ◽  
Myristate Acetate ◽  
Thyroid Cells ◽  
Pkc Inhibitor ◽  
Kinase C ◽  
Thyroid Cell Line ◽  
Rat Thyroid ◽  
Stimulation Of

In PC Cl3 cells, a continuous, fully differentiated rat thyroid cell line, P2Y2 purinoceptor activation provoked a transient increase of [Ca2+]i, followed by a decreasing sustained phase. The α and β1 protein kinase C (PKC) inhibitor Gö6976 decreased the rate of decrement to the basal [Ca2+]i level and increased the peak of Ca2+ entry of the P2Y2-provoked Ca2+transients. These effects of Gö 6976 were not caused by an increased permeability of the plasma membrane, since the Mn2+ and Ba2+ uptake were not changed by Gö 6976. Similarly, the Na+/Ca2+ exchanger was not implicated, since the rate of decrement to the basal [Ca2+]i level was equally decreased in physiological and Na+-free buffers, in the presence of Gö 6976. On the contrary, the activity of the sarcoplasmic–endoplasmic reticulum Ca2+ATPase (SERCA) 2b was profoundly affected by Gö 6976 since the drug was able to completely inhibit the stimulation of the SERCA 2b activity elicited by P2-purinergic agonists. Finally, the PKC activator phorbol myristate acetate had effects opposite to Gö 6976, in that it markedly increased the rate of decrement to the basal [Ca2+]i level after P2Y2 stimulation and also increased the activity of SERCA 2b. These results suggest that SERCA 2b plays a role in regulating the sustained phase of Ca2+ transients caused by P2Y2 stimulation.

Loss of adrenergic regulation of cAMP production in the FRTL-5 cell line

1986 ◽  
Vol 111 (1) ◽  
pp. 54-61 ◽  
Author(s):  
Maria Luisa Brandi ◽  
Carlo M. Rotella ◽  
Roberto Zonefrati ◽  
Roberto Toccafondi ◽  
Salvatore M. Aloj
Keyword(s):  
Primary Culture ◽  
Cell Line ◽  
Adrenergic Receptors ◽  
Primary Cultures ◽  
Human Thyroid ◽  
Camp Production ◽  
Thyroid Cells ◽  
Adrenergic Regulation ◽  
Rat Thyroid ◽  
Stimulation Of

Abstract. Rat thyroid cells in primary culture augment cAMP production when challenged with β-adrenergic agonists; at 10−5m the potency is isoproterenol > nor-epinephrine > epinephrine. In analogy with human thyroid cells, rat thyroid primary cultures display α-adrenergic-stimulated cGMP production which inhibits TSH and norepinephrine stimulation of cAMP. Adrenergic regulation of cyclic nucelotide production is lost in the cloned thyroid cell line of rat origin known as FRTL-5. Also the potentiating effect of phentolamine on TSH stimulation of cAMP production in thyroid primary cultures becomes an inhibitory one in the FRTL-5 cells. Neither 'soluble factors' nor contamination of other cell populations could account for the different behaviour of the primary culture and the cell line toward adrenergic regulation. The reported activation by norepinephrine of iodide efflux in FRTL-5 cells rules out the loss of specific adrenergic receptors in the FRTL-5 cells. It is proposed that the cloning of FRTL-5 cells from primary cultures causes an 'alteration' in the coupling of adrenergic receptors to the adenylate cyclase system. This alteration does not affect those mechansism of message transduction that do not involve cAMP as the signal.

Multiple pathways for cationic amino acid transport in rat thyroid epithelial cell line PC Cl3

2005 ◽  
Vol 288 (2) ◽  
pp. C290-C303 ◽  
Author(s):  
Tiziano Verri ◽  
Cinzia Dimitri ◽  
Sonia Treglia ◽  
Fabio Storelli ◽  
Stefania De Micheli ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Cell Line ◽  
Amino Acid Transport ◽  
Transport Systems ◽  
Residual Fraction ◽  
Acid Transport ◽  
Thyroid Cells ◽  
Cationic Amino Acid ◽  
Rat Thyroid

Information regarding cationic amino acid transport systems in thyroid is limited to Northern blot detection of y+LAT1 mRNA in the mouse. This study investigated cationic amino acid transport in PC cell line clone 3 (PC Cl3 cells), a thyroid follicular cell line derived from a normal Fisher rat retaining many features of normal differentiated follicular thyroid cells. We provide evidence that in PC Cl3 cells plasmalemmal transport of cationic amino acids is Na+ independent and occurs, besides diffusion, with the contribution of high-affinity, carrier-mediated processes. Carrier-mediated transport is via y+, y+L, and b0,+ systems, as assessed by l-arginine uptake and kinetics, inhibition of l-arginine transport by N-ethylmaleimide and neutral amino acids, and l-cystine transport studies. y+L and y+ systems account for the highest transport rate (with y+L > y+) and b0,+ for a residual fraction of the transport. Uptake data correlate to expression of the genes encoding for CAT-1, CAT-2B, 4F2hc, y+LAT1, y+LAT2, rBAT, and b0,+AT, an expression profile that is also shown by the rat thyroid gland. In PC Cl3 cells cationic amino acid uptake is under TSH and/or cAMP control (with transport increasing with increasing TSH concentration), and upregulation of CAT-1, CAT-2B, 4F2hc/y+LAT1, and rBAT/b0,+AT occurs at the mRNA level under TSH stimulation. Our results provide the first description of an expression pattern of cationic amino acid transport systems in thyroid cells. Furthermore, we provide evidence that extracellular l-arginine is a crucial requirement for normal PC Cl3 cell growth and that long-term l-arginine deprivation negatively influences CAT-2B expression, as it correlates to reduction of CAT-2B mRNA levels.

scholarly journals Importance and prospects for design of selective muscarinic agonists

2008 ◽  
pp. S39-S47
Author(s):  
J Jakubík ◽  
P Michal ◽  
E Machová ◽  
V Doležal
Keyword(s):  
Binding Sites ◽  
Natural Aging ◽  
Receptor Activation ◽  
Receptor Subtypes ◽  
Muscarinic Agonists ◽  
Amyloidogenic Processing ◽  
M1 Receptor ◽  
The Central Nervous System ◽  
M1 Agonist ◽  
Stimulation Of

There are five subtypes of muscarinic receptors that serve various important physiological functions in the central nervous system and the periphery. Mental functions like attention, learning, and memory are attributed to the muscarinic M1 subtype. These functions decline during natural aging and an early deficit is typical for Alzheimer s disease. In addition, stimulation of the M1 receptor increases non-amyloidogenic processing of the amyloid precursor protein and thus prevents accumulation of noxious beta-amyloid fragments. The selectivity of classical muscarinic agonists among receptor subtypes is very low due to the highly conserved nature of the orthosteric binding site among receptor subtypes. Herein we summarize some recent studies with the functionally-selective M1 agonist xanomeline that indicate complex pharmacological profile of this drug that includes interactions with and activation of receptor from both orthosteric and ectopic binding sites, and the time-dependent changes of ligand binding and receptor activation. These findings point to potential profitability of exploitation of ectopic ligands in the search for truly selective muscarinic receptor agonists.

Adhesion molecule expression by the FRTL-5 rat thyroid cell line

1991 ◽  
Vol 130 (3) ◽  
pp. 451-456 ◽  
Author(s):  
N. Tandon ◽  
C. Dinsdale ◽  
T. Tamatani ◽  
M. Miyasaka ◽  
A. P. Weetman
Keyword(s):  
Cell Line ◽  
Adhesion Molecule ◽  
Interleukin 1 ◽  
Intercellular Adhesion Molecule ◽  
Thyroid Cell ◽  
Lymphocyte Function ◽  
Intercellular Adhesion Molecule 1 ◽  
Thyroid Cells ◽  
Thyroid Cell Line ◽  
Rat Thyroid

ABSTRACT We have examined the expression and function of rat CD54, a homologue of human intercellular adhesion molecule-1 (ICAM-1), by the continuously growing rat thyroid cell line FRTL-5. Approximately 10% of FRTL-5 cells express CD54 under basal conditions and this is not influenced by thyrotrophin. Expression of CD54 is increased by cytokines (γ-interferon, tumour necrosis factor, interleukin-1) and by an activator of C-kinase, phorbol 12-myristate 13-acetate. Blocking ICAM-1 with a monoclonal antibody directed against this molecule significantly (P <0·01) reduced the binding of splenic lymphocytes to FRTL-5 cells but inhibition was consistently greater (P <0·01) in the presence of antibodies against a rat homologue of lymphocyte function-associated antigen-1, the receptor on T cells for ICAM-1. In no case was complete blocking of cluster formation observed. These results show that a pure line of rat thyroid cells can express an ICAM-1 homologue and this is directly enhanced by cytokines. Expression of this homologue is partially responsible for lymphocyte adhesion to thyroid cells, which is likely to be a major event in T cell recognition of thyroid antigens in autoimmune thyroiditis. Journal of Endocrinology (1991) 130, 451–456

scholarly journals Insulin and TSH promote growth in size of PC Cl3 rat thyroid cells, possibly via a pathway different from DNA synthesis: comparison with FRTL-5 cells

1999 ◽  
pp. 94-103 ◽  
Author(s):  
T Kimura ◽  
JE Dumont ◽  
A Fusco ◽  
J Golstein
Keyword(s):  
Protein Synthesis ◽  
Dna Replication ◽  
Dna Synthesis ◽  
Cell Lines ◽  
Insulin Action ◽  
Cell Mass ◽  
Thyroid Cell ◽  
Thyroid Cells ◽  
Rat Thyroid ◽  
Stimulation Of

In the rat thyroid cell lines PC Cl3, FRTL- 5 and WRT, proliferation is mainly regulated by insulin or IGF, and TSH. However, the mechanism regulating cell mass doubling prior to division is still unknown. Our laboratory has shown that in dog thyroid cells insulin promotes growth in size while TSH in the presence of insulin triggers DNA replication. In the absence of insulin, TSH has no effect on cell growth. In this report we investigated insulin action on both cell mass and DNA synthesis and its modulation by TSH and insulin in PC Cl3 and FRTL-5 cells. In PC Cl3 cells, insulin activated not only DNA synthesis but also protein synthesis and accumulation. Although TSH potentiated the stimulation of DNA synthesis induced by insulin, enhancement of protein synthesis by both agents was additive. All TSH effects were reproduced by forskolin. Similar effects were also obtained in FRTL-5 cells. This suggests that insulin and TSH, via cAMP, modulate both growth in size and DNA replication in these cell lines. Lovastatin, which blocks 3-hydroxy-3-methylglutaryl coenzyme A reductase, decreased the induction of DNA synthesis, but not of protein synthesis induced by insulin or TSH in PC Cl3 cells. In FRTL-5 cells, lovastatin reduced protein and DNA synthesis stimulated by insulin but not TSH-induced protein synthesis. Taking these data together, we propose that insulin and/or TSH both modulate cell mass doubling and DNA synthesis in these cell lines, presumably via different pathways, and that there are at least two pathways which regulate growth in size in FRTL-5 thyroid cells: one triggered by insulin, which is lovastatin sensitive, and the other activated by TSH, which is not sensitive to lovastatin.

The Role of 5–Hydroxytryptamine (5–HT) Receptor Subtypes and Plasticity in the 5–HT Systems in the Regulation of Nociceptive Sensitivity

Cephalalgia ◽  
1993 ◽  
Vol 13 (2) ◽  
pp. 75-85 ◽  
Author(s):  
Per Kristian Eide ◽  
Kjell Hole
Keyword(s):  
Receptor Activation ◽  
Term Treatment ◽  
Receptor Subtypes ◽  
Functional Changes ◽  
Nociceptive Sensitivity ◽  
Denervation Supersensitivity ◽  
Long Term Treatment ◽  
Stimulation Of

This review shows that the role of 5–hydroxytryptamine (5–HT) in the regulation of nociception depends on the 5–HT receptor subtypes involved and on long-term functional changes in the 5–HT receptors. Stimulation of the 5–HT 1 receptors, as well as of the 5–HT 2 and 5–HT 3 receptors, may reduce nociceptive sensitivity. In addition, activation of 5–HT 2 and 5–HT 3 receptors may also enhance nociceptive sensitivity. Up- or down-regulation of the 5–HT receptors may result in long-lasting changes, plasticity, in the 5–HT systems. Lesioning of 5–HT neurons induces denervation supersensitivity to 5–HT, and prolonged stimulation of 5–HT receptors may produce subsensitivity to 5–HT. In the spinal cord denervation supersensitivity to 5–HT may depend on reduced release of substance P (SP). An increase in the release of SP, on the other hand, may reduce the effects of 5–HT receptor activation. Long-term treatment with antidepressants which are used in clinical pain therapy appears to up-regulate the 5–HT 1 receptors and to down-regulate the 5–HT 2 receptors.

Differential effects of interleukin 1α and 1β on cultured human and rat thyroid epithelial cells

1990 ◽  
Vol 122 (4) ◽  
pp. 520-526 ◽  
Author(s):  
Å. Krogh Rasmussen ◽  
L. Kayser ◽  
K. Bech ◽  
U. Feldt-Rasmussen ◽  
H. Perrild ◽  
...  
Keyword(s):  
Cell Line ◽  
Interleukin 1 ◽  
Cell System ◽  
Human Thyroid ◽  
Thyroid Cell ◽  
Camp Production ◽  
Thyroid Cells ◽  
Thyroid Cell Line ◽  
Interleukin 1Α ◽  
Rat Thyroid

Abstract The effects of human recombinant interleukin 1α (20 pg/1-2 μg/l) and 1β (200 pg/1-20 μg/l) on two systems of thyroid cells have been compared. The thyroglobulin and cAMP secretion and the DNA content of human thyroid cells cultured in monolayer and of continuously grown rat thyroid cells, Fischer rat thyroid cell line have been studied. The growth of the rat thyroid cell line was inhibited by interleukin 1β (20 ng/1-20 μg/l), but not by interleukin 1α. None of the cytokines changed the cAMP production of the rat thyroid cells. In contrast, both cAMP production and thyroglobulin secretion were inhibited dose-dependently by the cytokines in human thyroid cells in secondary cultures. These results caution the interpretation and extrapolation of changes induced by interleukin 1 from one cell system to the other.

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