Glial-cell-line-derived neurotrophic factor induces nerve fibre formation in primary cultures of adrenal chromaffin cells

Calcium (Ca2+)-dependent endocytosis has been linked to preferential Ca2+ entry through the L-type (α1D, CaV1.3) of voltage-dependent Ca2+ channels (VDCCs). Considering that the Ca2+-dependent exocytotic release of neurotransmitters is mostly triggered by Ca2+ entry through N-(α1B, CaV2.2) or PQ-VDCCs (α1A, CaV2.1) and that exocytosis and endocytosis are coupled, the supposition that the different channel subtypes are specialized to control different cell functions is attractive. Here we have explored this hypothesis in primary cultures of bovine adrenal chromaffin cells where PQ channels account for 50% of Ca2+ current ( ICa), 30% for N channels, and 20% for L channels. We used patch-clamp and fluorescence techniques to measure the exo-endocytotic responses triggered by long depolarizing stimuli, in 1, 2, or 10 mM concentrations of extracellular Ca2+ ([Ca2+]e). Exo-endocytotic responses were little affected by ω-conotoxin GVIA (N channel blocker), whereas ω-agatoxin IVA (PQ channel blocker) caused 80% blockade of exocytosis as well as endocytosis. In contrast, nifedipine (L channel blocker) only caused 20% inhibition of exocytosis but as much as 90% inhibition of endocytosis. Conversely, FPL67146 (an activator of L VDCCs) notably augmented endocytosis. Photoreleased caged Ca2+ caused substantially smaller endocytotic responses compared with those produced by K+ depolarization. Using fluorescence antibodies, no colocalization between L, N, or PQ channels with clathrin was found; a 20–30% colocalization was found between dynamin and all three channel antibodies. This is incompatible with the view that L channels are coupled to the endocytotic machine. Data rather support a mechanism implying the different inactivation rates of L (slow-inactivating) and N/PQ channels (fast-inactivating). Thus a slow but more sustained Ca2+ entry through L channels could be a requirement to trigger endocytosis efficiently, at least in bovine chromaffin cells.

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Effects of cordycepin (3'-deoxyadenosine) on 8-Br cAMP-mediated induction of tyrosine hydroxylase (TH) in primary cultures of bovine adrenal chromaffin cells

The Japanese Journal of Pharmacology ◽

10.1016/s0021-5198(19)54435-3 ◽

1981 ◽

Vol 31 ◽

pp. 90

Author(s):

Konosuke Kumakura

Keyword(s):

Tyrosine Hydroxylase ◽

Chromaffin Cells ◽

Primary Cultures ◽

Adrenal Chromaffin Cells ◽

Adrenal Chromaffin

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Enkephalin congeners and precursors are synthesized and released by primary cultures of adrenal chromaffin cells

Life Sciences ◽

10.1016/0024-3205(81)90161-2 ◽

1981 ◽

Vol 28 (7) ◽

pp. 781-789 ◽

Cited By ~ 43

Author(s):

Jean Rossier ◽

Deanne M. Dean ◽

Bruce G. Livett ◽

Sidney Udenfriend

Keyword(s):

Chromaffin Cells ◽

Primary Cultures ◽

Adrenal Chromaffin Cells ◽

Adrenal Chromaffin

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Stimulation of Formation of Inositol Phosphates in Primary Cultures of Bovine Adrenal Chromaffin Cells by Angiotensin II, Histamine, Bradykinin, and Carbachol

Journal of Neurochemistry ◽

10.1111/j.1471-4159.1988.tb01085.x ◽

1988 ◽

Vol 51 (2) ◽

pp. 634-641 ◽

Cited By ~ 88

Author(s):

Robin Plevin ◽

Michael R. Boarder

Keyword(s):

Angiotensin Ii ◽

Chromaffin Cells ◽

Inositol Phosphates ◽

Primary Cultures ◽

Adrenal Chromaffin Cells ◽

Adrenal Chromaffin ◽

Stimulation Of

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

Biochemical Journal ◽

10.1042/bj2510397 ◽

1988 ◽

Vol 251 (2) ◽

pp. 397-403 ◽

Cited By ~ 26

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.

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Gabapentin Inhibits Catecholamine Release from Adrenal Chromaffin Cells

Anesthesiology ◽

10.1097/aln.0b013e31825153ea ◽

2012 ◽

Vol 116 (5) ◽

pp. 1013-1024 ◽

Cited By ~ 14

Author(s):

Robert D. Todd ◽

Sarah M. McDavid ◽

Rebecca L. Brindley ◽

Mark L. Jewell ◽

Kevin P. M. Currie

Keyword(s):

Calcium Channel ◽

Chromaffin Cells ◽

Single Cells ◽

Primary Cultures ◽

Catecholamine Release ◽

Fluorescent Imaging ◽

Catecholamine Secretion ◽

Adrenal Chromaffin Cells ◽

Adrenal Chromaffin ◽

Channel Currents

Background Gabapentin is most commonly prescribed for chronic pain, but acute perioperative effects, including preemptive analgesia and hemodynamic stabilization, have been reported. Adrenal chromaffin cells are a widely used model to investigate neurosecretion, and adrenal catecholamines play important physiologic roles and contribute to the acute stress response. However, the effects of gabapentin on adrenal catecholamine release have never been tested. Methods Primary cultures of bovine adrenal chromaffin cells were treated with gabapentin or vehicle for 18-24 h. The authors quantified catecholamine secretion from dishes of cells using high-performance liquid chromatography and resolved exocytosis of individual secretory vesicles from single cells using carbon fiber amperometry. Voltage-gated calcium channel currents were recorded using patch clamp electrophysiology and intracellular [Ca2+] using fluorescent imaging. Results Gabapentin produced statistically significant reductions in catecholamine secretion evoked by cholinergic agonists (24 ± 3%, n = 12) or KCl (16 ± 4%, n = 8) (mean ± SEM) but did not inhibit Ca2+ entry or calcium channel currents. Amperometry (n = 51 cells) revealed that gabapentin inhibited the number of vesicles released upon stimulation, with no change in quantal size or kinetics of these unitary events. Conclusions The authors show Ca2+ entry was not inhibited by gabapentin but was less effective at triggering vesicle fusion. The work also demonstrates that chromaffin cells are a useful model for additional investigation of the cellular mechanism(s) by which gabapentin controls neurosecretion. In addition, it identifies altered adrenal catecholamine release as a potential contributor to some of the beneficial perioperative effects of gabapentin.

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