scholarly journals Phosphatidylinositol-4,5-Biphosphate (PI(4,5)P2) Is Required for Rapid Endocytosis in Chromaffin Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-6
Author(s):  
Fouad Azizi
Keyword(s):  
Neurotransmitter Release ◽  
Chromaffin Cells ◽  
Inhibitory Effect ◽  
Adrenal Chromaffin Cells ◽  
Patch Clamp Recording ◽  
Vesicle Recycling ◽  
Patch Pipette ◽  
Cell Membrane Capacitance ◽  
Adrenal Chromaffin ◽  
Phosphatidylinositol 4

Objective. Phosphoinositides play a regulatory role in clathrin-mediated endocytosis. However, their involvement in clathrin-independent endocytosis termed rapid endocytosis (RE), which is the mode of vesicle recycling during neurotransmitter release by transient fusion (known as kiss-and-run), has not been investigated. Here, we used patch-clamp recording of whole-cell membrane capacitance in adrenal chromaffin cells (ACC) to monitor changes of RE kinetics in response to pharmacological alteration of phosphatidylinositol-4,5-biphosphate (PI(4,5)P2) level by phenylarsine oxide (PAO) or antibody against phosphatidylinositol 4-kinase (AbPI4K). Results. We found that PAO and AbPI4K significantly abrogated RE kinetics. Infusion of PI(4,5)P2 through the patch pipette potentiated RE kinetics and reversed PAO- and AbPI4K-induced blockade of RE. Similarly, the application of the bifunctional thiol dithiothreitol (DTT) to PAO-treated cells completely prevented the inhibitory effect of PAO on RE. These findings indicate that PI(4,5)P2 is implicated in the signaling (mechanistic) process of RE in ACC.

scholarly journals Exocytosis from permeabilized bovine adrenal chromaffin cells is differently modulated by guanosine 5′-[γ-thio]triphosphate and guanosine 5′-[β γ-imido]triphosphate. Evidence for the involvement of various guanine nucleotide-binding proteins

1992 ◽  
Vol 284 (2) ◽  
pp. 321-326 ◽  
Author(s):  
G Ahnert-Hilger ◽  
U Wegenhorst ◽  
B Stecher ◽  
K Spicher ◽  
W Rosenthal ◽  
...  
Keyword(s):  
G Protein ◽  
Chromaffin Cells ◽  
Inhibitory Effect ◽  
Adrenal Chromaffin Cells ◽  
Prolonged Incubation ◽  
Permeabilized Cells ◽  
Alpha Subunits ◽  
Cytoplasmic Material ◽  
Streptolysin O ◽  
Adrenal Chromaffin

1. In bovine adrenal chromaffin cells made permeable either to molecules less than or equal to 3 kDa with alphatoxin or to proteins less than or equal to 150 kDa with streptolysin O, the GTP analogues guanosine 5′-[beta gamma-imido]triphosphate (p[NH]ppG) and guanosine 5′-[gamma-thio]triphosphate (GTP[S]) differently modulated Ca(2+)-stimulated exocytosis. 2. In alphatoxin-permeabilized cells, p[NH]ppG up to 20 microM activated Ca(2+)-stimulated exocytosis. Higher concentrations had little or no effect. At a free Ca2+ concentration of 5 microM, 7 microM-p[NH]ppG stimulated exocytosis 6-fold. Increasing the free Ca2+ concentration reduced the effect of p[NH]ppG. Pretreatment of the cells with pertussis toxin prevented the activation of the Ca(2+)-stimulated exocytosis by p[NH]ppG. 3. In streptolysin O-permeabilized cells, p[NH]ppG did not activate, but rather inhibited Ca(2+)-dependent catecholamine release under all conditions studied. In the soluble cytoplasmic material that escaped during permeabilization with streptolysin O, different G-protein alpha-subunits were detected using an appropriate antibody. Around 15% of the cellular alpha-subunits were detected in the supernatant of permeabilized control cells. p[NH]ppG or GTP[S] stimulated the release of alpha-subunits 2-fold, causing a loss of about 30% of the cellular G-protein alpha-subunits under these conditions. Two of the alpha-subunits in the supernatant belonged to the G(o) type, as revealed by an antibody specific for G(o) alpha. 4. GTP[S], when present alone during stimulation with Ca2+, activated exocytosis in a similar manner to p[NH]ppG. Upon prolonged incubation, GTP[S], in contrast to p[NH]ppG, inhibited Ca(2+)-induced exocytosis from cells permeabilized by either of the pore-forming toxins. This effect was resistant to pertussin toxin. 5. The p[NH]ppG-induced activation of Ca(2+)-stimulated release from alphatoxin-permeabilized chromaffin cells may be attributed to one of the heterotrimeric G-proteins lost during permeabilization with streptolysin O. The inhibitory effect of GTP[S] on exocytosis is apparently not mediated by G-protein alpha-subunits, but by another GTP-dependent process still occurring after permeabilization with streptolysin O.

scholarly journals Relationship between arachidonic acid release and Ca2+-dependent exocytosis in digitonin-permeabilized bovine adrenal chromaffin cells

1990 ◽  
Vol 271 (3) ◽  
pp. 571-574 ◽  
Author(s):  
A Morgan ◽  
R D Burgoyne
Keyword(s):  
Arachidonic Acid ◽  
Phospholipase A2 ◽  
Chromaffin Cells ◽  
Nordihydroguaiaretic Acid ◽  
Inhibitory Effect ◽  
Arachidonic Acid Release ◽  
Adrenal Chromaffin Cells ◽  
Protein Kinase C Inhibitor ◽  
Acid Release ◽  
Adrenal Chromaffin

The relationship between Ca2(+)-dependent arachidonic acid release and exocytosis from digitonin-permeabilized bovine adrenal chromaffin cells was investigated. The phospholipase A2 inhibitors mepacrine, nordihydroguaiaretic acid and indomethacin had no effect on either arachidonic acid release or secretion. The phospholipase A2 activator melittin had no effect on secretion. The specific diacylglycerol lipase inhibitor RG80267 had no effect on secretion, but decreased basal arachidonic acid release to such an extent that the level of arachidonic acid in treated cells in response to 10 microM-Ca2+ was equivalent to that of control cells in the absence of Ca2+. Staurosporine, a protein kinase C inhibitor, was found to abolish Ca2(+)-dependent arachidonic acid release completely, but had only a slight inhibitory effect on Ca2(+)-dependent secretion. It is concluded that arachidonic acid is not essential for Ca2(+)-dependent exocytosis in adrenal chromaffin cells.

Difference in neurotransmitter release induced by calcium channel and acetylcholine receptor activation in adrenal chromaffin cells

2006 ◽  
Vol 99 (2-3) ◽  
pp. 1
Author(s):  
Elena A. Lukyanetz ◽  
Oleg M. Pochynyuk ◽  
Oleg L. Zaika ◽  
Larisa M. Koval ◽  
Elena N. Yavorskaya ◽  
...  
Keyword(s):  
Calcium Channel ◽  
Acetylcholine Receptor ◽  
Neurotransmitter Release ◽  
Chromaffin Cells ◽  
Receptor Activation ◽  
Adrenal Chromaffin Cells ◽  
Adrenal Chromaffin

scholarly journals Functional shift from muscarinic to nicotinic cholinergic receptors involved in inositol trisphosphate and cyclic GMP accumulation during the primary culture of adrenal chromaffin cells

1988 ◽  
Vol 251 (2) ◽  
pp. 397-403 ◽  
Author(s):  
T Nakaki ◽  
N Sasakawa ◽  
S Yamamoto ◽  
R Kato
Keyword(s):  
Primary Culture ◽  
Cyclic Gmp ◽  
Chromaffin Cells ◽  
Primary Cultures ◽  
Cholinergic Receptor ◽  
Inhibitory Effect ◽  
Cholinergic Receptors ◽  
Receptor Subtypes ◽  
Adrenal Chromaffin Cells ◽  
Adrenal Chromaffin

Specificities of cholinergic receptors for the accumulation of inositol trisphosphates (InsP3) and cyclic GMP and mobilization of intracellular Ca2+ in relation to culture periods were investigated in primary cultures of bovine adrenal chromaffin cells. At 0.5 day in culture, muscarine, a specific agonist for muscarinic receptors, caused a greater effect on intracellular Ca2+ mobilization and the accumulation of Ins(1,3,4)P3 than did the nicotinic-specific agonist nicotine. On the contrary, at 5 days, nicotine produced a greater effect on the accumulation of Ins(1,3,4)P3 and intracellular calcium mobilization than did muscarine. Furthermore, at 0.5 day, the muscarinic antagonist atropine strongly inhibited the increase in InsP3 accumulation that was induced by the nonspecific agonist carbachol, whereas at 5 days the inhibitory effect of atropine was greatly lowered. On the other hand, the nicotinic receptor antagonists hexamethonium and d-tubocurarine showed a much higher inhibitory potency at 5 days compared with 0.5 day in culture. Cholinergic receptor subtypes involved in cyclic GMP accumulation showed functional shifts similar to those in InsP3 formation. Binding experiments with a muscarinic ligand excluded the possibility that the reduction in muscarinic effects on InsP3 and cyclic GMP formation and intracellular Ca2+ mobilization were due to disappearance of the muscarinic receptor itself. These data show that cholinergic receptors linked to the accumulation of InsP3 and cyclic GMP and Ca2+ mobilization functionally shift from muscarinic to nicotinic during primary culture of adrenal chromaffin cells.

Voltage-Dependent, Pertussis Toxin Insensitive Inhibition of Calcium Currents by Histamine in Bovine Adrenal Chromaffin Cells

2000 ◽  
Vol 83 (3) ◽  
pp. 1435-1442 ◽  
Author(s):  
Kevin P. M. Currie ◽  
Aaron P. Fox
Keyword(s):  
G Proteins ◽  
Pertussis Toxin ◽  
Chromaffin Cells ◽  
Calcium Influx ◽  
Adrenal Chromaffin Cells ◽  
Pipette Solution ◽  
Patch Pipette ◽  
Calcium Buffering ◽  
Voltage Dependent ◽  
Adrenal Chromaffin

Histamine is a known secretagogue in adrenal chromaffin cells. Activation of G-protein linked H1 receptors stimulates phospholipase C, which generates inositol trisphosphate leading to release of intracellular calcium stores and stimulation of calcium influx through store operated and other channels. This calcium leads to the release of catecholamines. In chromaffin cells, the main physiological trigger for catecholamine release is calcium influx through voltage-gated calcium channels ( I Ca). Therefore, these channels are important targets for the regulation of secretion. In particular N- and P/Q-type I Ca are subject to inhibition by transmitter/hormone receptor activation of heterotrimeric G-proteins. However, the direct effect of histamine on I Ca in chromaffin cells is unknown. This paper reports that histamine inhibited I Cain cultured bovine adrenal chromaffin cells and this response was blocked by the H1 antagonist mepyramine. With high levels of calcium buffering in the patch pipette solution (10 mM EGTA), histamine slowed the activation kinetics and inhibited the amplitude of I Ca. A conditioning prepulse to +100 mV reversed the kinetic slowing and partially relieved the inhibition. These features are characteristic of a membrane delimited, voltage-dependent pathway which is thought to involve direct binding of G-protein βγ subunits to the Ca channels. However, unlike virtually every other example of this type of inhibition, the response to histamine was not blocked by pretreating the cells with pertussis toxin (PTX). The voltage-dependent, PTX insensitive inhibition produced by histamine was modest compared with the PTX sensitive inhibition produced by ATP (28% vs. 53%). When histamine and ATP were applied concomitantly there was no additivity of the inhibition beyond that produced by ATP alone (even though the agonists appear to activate distinct G-proteins) suggesting that the inhibition produced by ATP is maximal. When experiments were carried out under conditions of low levels of calcium buffering in the patch pipette solution (0.1 mM EGTA), histamine inhibited I Ca in some cells using an entirely voltage insensitive pathway. We demonstrate that activation of PTX insensitive G-proteins (most likely Gq) by H1 receptors inhibits I Ca. This may represent a mechanism by which histamine exerts inhibitory (in addition to previously identified stimulatory) effects on catecholamine release.

Morphological Observations on the Granule Types in Rabbit Extra-Adrenal Chromaffin Cells.

1975 ◽  
Vol 33 ◽  
pp. 318-319
Author(s):  
Joe A. Mascorro ◽  
Robert D. Yates
Keyword(s):  
Chromaffin Cells ◽  
Sodium Phosphate ◽  
Ganglion Cells ◽  
Ultrastructural Level ◽  
Osmic Acid ◽  
Adrenal Chromaffin Cells ◽  
Potassium Iodate ◽  
Perfusion Fluid ◽  
New Zealand White Rabbits ◽  
Adrenal Chromaffin

Extra-adrenal chromaffin organs (abdominal paraganglia) constitute rich sources of catecholamines. It is believed that these bodies contain norepinephrine exclusively. However, the present workers recently observed epinephrine type granules in para- ganglion cells. This report investigates catecholamine containing granules in rabbit paraganglia at the ultrastructural level.New Zealand white rabbits (150-170 grams) were anesthetized with 50 mg/kg Nembutal (IP) and perfused with 3% glutaraldehyde buffered with 0.2M sodium phosphate, pH 7.3. The retroperitoneal tissue blocks were removed and placed in perfusion fluid for 4 hours. The abdominal paraganglia were dissected from the blocks, diced, washed in phosphate buffer and fixed in 1% osmic acid buffered with phosphate. In other animals, the glutaraldehyde perfused tissue blocks were immersed for 1 hour in 3% glutaraldehyde/2.5% potassium iodate buffered as before. The paraganglia were then diced, separated into two vials and washed in the buffer. A portion of this tissue received osmic acid fixation.

Sign in / Sign up

Export Citation Format

Share Document