Functional mitochondria are required for O2 but not CO2 sensing in immortalized adrenomedullary chromaffin cells

2008 ◽  
Vol 294 (4) ◽  
pp. C945-C956 ◽  
Author(s):  
J. Buttigieg ◽  
S. T. Brown ◽  
M. Lowe ◽  
M. Zhang ◽  
C. A. Nurse
Keyword(s):  
Complex I ◽  
Chromaffin Cells ◽  
Chromaffin Cell ◽  
Carbonic Anhydrase Ii ◽  
Adrenal Chromaffin Cells ◽  
Wild Type ◽  
Mitochondrial Complex ◽  
Adrenal Chromaffin Cell ◽  
Adrenal Chromaffin ◽  
First Time

Catecholamine (CAT) release from adrenomedullary chromaffin cells (AMC) in response to stressors such as low O2 (hypoxia) and elevated CO2/H+ is critical during adaptation of the newborn to extrauterine life. Using a surrogate model based on a v -myc immortalized adrenal chromaffin cell line (i.e., MAH cells), combined with genetic perturbation of mitochondrial function, we tested the hypothesis that functional mitochondria are required for O2 sensing. Wild-type MAH cells responded to both hypoxia and increased CO2 (hypercapnia) with K+ current inhibition and membrane depolarization. Additionally, these stimuli caused a rise in cytosolic Ca2+ and CAT secretion, determined by fura-2 spectrofluorimetry and carbon fiber amperometry, respectively. In contrast, mitochondria-deficient (ρ0) MAH cells were hypoxia insensitive, although responses to hypercapnia and expression of several markers, including carbonic anhydrase II, remained intact. Rotenone (1 μM), a mitochondrial complex I blocker known to mimic and occlude the effects of hypoxia in primary AMC, was effective in wild-type but not ρ0 MAH cells. These data demonstrate that functional mitochondria are involved in hypoxia-sensing by adrenal chromaffin cells. We also show for the first time that, like their neonatal chromaffin cell counterparts, MAH cells are CO2 sensors; however, this property is independent of functional mitochondria.

Development of chromaffin cells depends on MASH1 function

Development ◽  
2002 ◽  
Vol 129 (20) ◽  
pp. 4729-4738 ◽  
Author(s):  
Katrin Huber ◽  
Barbara Brühl ◽  
François Guillemot ◽  
Eric N. Olson ◽  
Uwe Ernsberger ◽  
...  
Keyword(s):  
Adrenal Medulla ◽  
Chromaffin Cells ◽  
Adrenal Glands ◽  
Sympathetic Neurons ◽  
Cell Lineage ◽  
Neuroendocrine Differentiation ◽  
Sympathetic Ganglia ◽  
Adrenal Chromaffin Cells ◽  
Wild Type ◽  
Adrenal Chromaffin

The sympathoadrenal (SA) cell lineage is a derivative of the neural crest (NC), which gives rise to sympathetic neurons and neuroendocrine chromaffin cells. Signals that are important for specification of these two types of cells are largely unknown. MASH1 plays an important role for neuronal as well as catecholaminergic differentiation. Mash1 knockout mice display severe deficits in sympathetic ganglia, yet their adrenal medulla has been reported to be largely normal suggesting that MASH1 is essential for neuronal but not for neuroendocrine differentiation. We show now that MASH1 function is necessary for the development of the vast majority of chromaffin cells. Most adrenal medullary cells in Mash1–/– mice identified by Phox2b immunoreactivity, lack the catecholaminergic marker tyrosine hydroxylase. Mash1 mutant and wild-type mice have almost identical numbers of Phox2b-positive cells in their adrenal glands at embryonic day (E) 13.5; however, only one-third of the Phox2b-positive adrenal cell population seen in Mash1+/+ mice is maintained in Mash1–/– mice at birth. Similar to Phox2b, cells expressing Phox2a and Hand2 (dHand) clearly outnumber TH-positive cells. Most cells in the adrenal medulla of Mash1–/– mice do not contain chromaffin granules, display a very immature, neuroblast-like phenotype, and, unlike wild-type adrenal chromaffin cells, show prolonged expression of neurofilament and Ret comparable with that observed in wild-type sympathetic ganglia. However, few chromaffin cells in Mash1–/– mice become PNMT positive and downregulate neurofilament and Ret expression. Together, these findings suggest that the development of chomaffin cells does depend on MASH1 function not only for catecholaminergic differentiation but also for general chromaffin cell differentiation.

Gaseous Transmitter Regulation of Catecholamine Secretion by hypoxia in Murine Adrenal Chromaffin Cells

2021 ◽  
Author(s):  
Anna Gridina ◽  
Xiaoyu Su ◽  
Shakil A. Khan ◽  
Ying-Jie Peng ◽  
Benjamin L Wang ◽  
...  
Keyword(s):  
Nervous System ◽  
Protein Kinase ◽  
Guanylyl Cyclase ◽  
Chromaffin Cells ◽  
Soluble Guanylyl Cyclase ◽  
Protein Kinase G ◽  
Adrenal Chromaffin Cells ◽  
Wild Type ◽  
Genetic Deletion ◽  
Adrenal Chromaffin

Emerging evidence suggests that gaseous molecules, carbon monoxide (CO) and hydrogen sulfide (H2S) generated by heme oxygenase-(HO)-2 and cystathionine γ-lyase (CSE), respectively, function as transmitters in the nervous system. Present study examined the roles of CO and H2S in hypoxia-induced catecholamine (CA) release from adrenal medullary chromaffin cells (AMC). Studies were performed on AMC from adult (≥6 weeks of age) wild type (WT), HO-2 null, CSE null and HO-2/CSE double null mice of either gender. CA secretion was determined by carbon fiber amperometry and [Ca2+]i by microflurometry using Fura-2. HO-2- and CSE immunoreactivities were seen in WT AMC, which were absent in HO-2 and CSE null mice. Hypoxia (medium pO2 30-38 mmHg) evoked CA release and elevated [Ca2+]i. The magnitude of hypoxic response was greater in HO-2 null mice and in HO inhibitor treated WT AMC compared to controls. H2S levels were elevated in HO-2 null AMC. Either pharmacological inhibition or genetic deletion of CSE prevented the augmented hypoxic responses of HO-2 null AMC and H2S donor rescued AMC responses to hypoxia in HO-2/CSE double null mice. CORM-3, a CO donor, prevented the augmented hypoxic responses in WT and HO-2 null AMC. CO donor reduced H2S levels in WT AMC. The effects of CO donor were blocked by either ODQ or 8pCT, inhibitors of soluble guanylyl cyclase (SGC) or protein kinase G, respectively. These results suggest that HO-2-derived CO inhibits hypoxia-evoked CA secretion from adult murine AMC involving soluble guanylyl cyclase (SGC)-protein kinase G (PKG)-dependent regulation of CSE- derived H2S.

NGF-like trophic support from peripheral nerve for grafted rhesus adrenal chromaffin cells

1990 ◽  
Vol 73 (3) ◽  
pp. 418-428 ◽  
Author(s):  
Jeffrey H. Kordower ◽  
Massimo S. Fiandaca ◽  
Mary F. D. Notter ◽  
John T. Hansen ◽  
Don M. Gash
Keyword(s):  
Tyrosine Hydroxylase ◽  
Adrenal Medulla ◽  
Sural Nerve ◽  
Chromogranin A ◽  
Chromaffin Cells ◽  
Chromaffin Cell ◽  
Adrenal Chromaffin Cells ◽  
Content Type ◽  
Adrenal Chromaffin ◽  
Fine Print

✓ Autopsy results on patients and corresponding studies in nonhuman primates have revealed that autografts of adrenal medulla into the striatum, used as a treatment for Parkinson's disease, do not survive well. Because adrenal chromaffin cell viability may be limited by the low levels of available nerve growth factor (NGF) in the striatum, the present study was conducted to determine if transected peripheral nerve segments could provide sufficient levels of NGF to enhance chromaffin cell survival in vitro and in vivo. Aged female rhesus monkeys, rendered hemiparkinsonian by the drug MPTP (n-methyl-4-phenyl-1,2,3,6 tetrahydropyridine), received autografts into the striatum using a stereotactic approach, of either sural nerve or adrenal medulla, or cografts of adrenal medulla and sural nerve (three animals in each group). Cell cultures were established from tissue not used in the grafts. Adrenal chromaffin cells either cocultured with sural nerve segments or exposed to exogenous NGF differentiated into a neuronal phenotype. Chromaffin cell survival, when cografted with sural nerve into the striatum, was enhanced four- to eightfold from between 8000 and 18,000 surviving cells in grafts of adrenal tissue only up to 67,000 surviving chromaffin cells in cografts. In grafts of adrenal tissue only, the implant site consisted of an inflammatory focus. Surviving chromaffin cells, which could be identified by both chromogranin A and tyrosine hydroxylase staining, retained their endocrine phenotype. Cografted chromaffin cells exhibited multipolar neuritic processes and numerous chromaffin granules, and were also immunoreactive for tyrosine hydroxylase and chromogranin A. Blood vessels within the graft were fenestrated, indicating that the blood-brain barrier was not intact. Additionally, cografted chromaffin cells were observed in a postsynaptic relationship with axon terminals from an undetermined but presumably a host origin.

scholarly journals Analysis of mice carrying targeted mutations of the glucocorticoid receptor gene argues against an essential role of glucocorticoid signalling for generating adrenal chromaffin cells

Development ◽  
1999 ◽  
Vol 126 (13) ◽  
pp. 2935-2944 ◽  
Author(s):  
S. Finotto ◽  
K. Krieglstein ◽  
A. Schober ◽  
F. Deimling ◽  
K. Lindner ◽  
...  
Keyword(s):  
Glucocorticoid Receptor ◽  
Chromaffin Cells ◽  
Essential Role ◽  
Adrenal Chromaffin Cells ◽  
Wild Type ◽  
Sympathetic Neurones ◽  
Adrenal Chromaffin ◽  
Deficient Mice

Molecular mechanisms underlying the generation of distinct cell phenotypes is a key issue in developmental biology. A major paradigm of determination of neural cell fate concerns the development of sympathetic neurones and neuroendocrine chromaffin cells from a common sympathoadrenal (SA) progenitor cell. Two decades of in vitro experiments have suggested an essential role of glucocorticoid receptor (GR)-mediated signalling in generating chromaffin cells. Targeted mutation of the GR should consequently abolish chromaffin cells. The present analysis of mice lacking GR gene product demonstrates that animals have normal numbers of adrenal chromaffin cells. Moreover, there are no differences in terms of apoptosis and proliferation or in expression of several markers (e.g. GAP43, acetylcholinesterase, adhesion molecule L1) of chromaffin cells in GR-deficient and wild-type mice. However, GR mutant mice lack the adrenaline-synthesizing enzyme PNMT and secretogranin II. Chromaffin cells of GR-deficient mice exhibit the typical ultrastructural features of this cell phenotype, including the large chromaffin granules that distinguish them from sympathetic neurones. Peripherin, an intermediate filament of sympathetic neurones, is undetectable in chromaffin cells of GR mutants. Finally, when stimulated with nerve growth factor in vitro, identical proportions of chromaffin cells from GR-deficient and wild-type mice extend neuritic processes. We conclude that important phenotypic features of chromaffin cells that distinguish them from sympathetic neurones develop normally in the absence of GR-mediated signalling. Most importantly, chromaffin cells in GR-deficient mice do not convert to a neuronal phenotype. These data strongly suggest that the dogma of an essential role of glucocorticoid signalling for the development of chromaffin cells must be abandoned.

Effects of substance P on the long-term regulation of tyrosine hydroxylase activity and catecholamine levels in cultured adrenal chromaffin cells

1986 ◽  
Vol 64 (12) ◽  
pp. 1548-1555 ◽  
Author(s):  
P. Boksa
Keyword(s):  
Tyrosine Hydroxylase ◽  
Substance P ◽  
Chromaffin Cells ◽  
Chromaffin Cell ◽  
Splanchnic Nerve ◽  
Adrenal Chromaffin Cells ◽  
Hydroxylase Activity ◽  
Tyrosine Hydroxylase Activity ◽  
Adrenal Chromaffin

Acetylcholine, released from splanchnic nerve terminals innervating adrenal chromaffin cells, is known to increase synthesis of adrenal tyrosine hydroxylase, the rate-limiting enzyme in catecholamine synthesis. The neuropeptide substance P is also present in the splanchnic nerve innervating the adrenal medulla, and this study examined whether substance P has any long-term effects on tyrosine hydroxylase activity and catecholamine levels in cultures of adult bovine adrenal chromaffin cells. When cultures were incubated for 3 days with substance P and carbachol, a cholinergic agonist, substance P (10−6 M, and greater) completely inhibited the increase in tyrosine hydroxylase activity normally induced by carbachol. Long-term stimulation with carbachol also depleted endogenous catecholamines from the cells and substance P prevented this carbachol-induced depletion of catecholamine content. Substance P by itself, in the absence of carbachol, had only a slight effect on tyrosine hydroxylase activity. 8-Bromoadenosine 3′:5′-cyclic monophosphate, an analogue of adenosine 3′:5′-cyclic monophosphate, also increases tyrosine hydroxylase activity in chromaffin cells; however, substance P had no effect on the increase in tyrosine hydroxylase activity induced by this analogue. These results indicate that substance P's effects are relatively specific for the carbachol-induced increased in tyrosine hydroxylase activity and that the primary site of action of substance P is not a site common to the mechanism of tyrosine hydroxylase induction by carbachol and 8-bromoadenosine 3′:5′-cyclic monophosphate. During long-term incubation of chromaffin cell cultures, substance P inhibited the carbachol-induced increase in tyrosine hydroxylase activity only if peptidase inhibitors, bacitracin and captopril, were included in the medium, suggesting that chromaffin cell peptidases may be capable of terminating substance P's actions in these cells. The peptidase inhibitors bacitracin and captopril alone increased tyrosine hydroxylase activity and depleted catecholamines from the cells. The results suggest that substance P, released either from the splanchnic nerve or from the chromaffin cells themselves, may play a role in the long-term regulation of tyrosine hydroxylase activity and catecholamine levels in the mature adrenal chromaffin cell.

scholarly journals Conditional Inactivation of Glucocorticoid Receptor Gene in Dopamine-β-Hydroxylase Cells Impairs Chromaffin Cell Survival

Endocrinology ◽  
2008 ◽  
Vol 150 (4) ◽  
pp. 1775-1781 ◽  
Author(s):  
Rosanna Parlato ◽  
Christiane Otto ◽  
Jan Tuckermann ◽  
Stefanie Stotz ◽  
Sylvia Kaden ◽  
...  
Keyword(s):  
Glucocorticoid Receptor ◽  
Chromaffin Cells ◽  
Chromaffin Cell ◽  
Apoptotic Cell ◽  
Receptor Gene ◽  
Adrenal Chromaffin Cells ◽  
Glucocorticoid Receptor Gene ◽  
Conditional Inactivation ◽  
Adrenal Chromaffin

Glucocorticoid hormones (GCs) have been thought to determine the fate of chromaffin cells from sympathoadrenal progenitor cells. The analysis of mice carrying a germ line deletion of the glucocorticoid receptor (GR) gene has challenged these previous results because the embryonic development of adrenal chromaffin cells is largely unaltered. In the present study, we have analyzed the role of GC-dependent signaling in the postnatal development of adrenal chromaffin cells by conditional inactivation of the GR gene in cells expressing dopamine-β-hydroxylase, an enzyme required for the synthesis of noradrenaline and adrenaline. These mutant mice are viable, allowing to study whether in the absence of GC signaling further development of the adrenal medulla is affected. Our analysis shows that the loss of GR leads not only to the loss of phenylethanolamine-N-methyl-transferase expression and, therefore, to inhibition of adrenaline synthesis, but also to a dramatic reduction in the number of adrenal chromaffin cells. We provide evidence that increased apoptotic cell death is the main consequence of GR loss. These findings define the essential role of GCs for survival of chromaffin cells and underscore the specific requirement of GCs for adrenergic chromaffin cell differentiation and maintenance.

Permeation and inactivation by calcium and manganese of bovine adrenal chromaffin cell calcium channels

1992 ◽  
Vol 263 (4) ◽  
pp. C818-C824 ◽  
Author(s):  
R. I. Fonteriz ◽  
J. Garcia-Sancho ◽  
L. Gandia ◽  
M. G. Lopez ◽  
A. G. Garcia
Keyword(s):  
Chromaffin Cells ◽  
Chromaffin Cell ◽  
Divalent Cations ◽  
Receptor Activation ◽  
Adrenal Chromaffin Cell ◽  
High K ◽  
Bovine Chromaffin Cells ◽  
Adrenal Chromaffin ◽  
Ca2 Channels ◽  
Stimulation Of

Stimulation of fura-2-loaded bovine chromaffin cells with the nicotinic agonist 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP; 10 microM) or depolarization with high [K+] (50 mM) accelerated the entry of both Ca2+ and Mn2+, used here as a Ca2+ surrogate for Ca2+ channels. Removal of extracellular Na+ prevented the effects of DMPP but did not modify the effects of K+, indicating that Na+ is necessary for coupling of Ca2+ entry to the nicotinic receptor activation and that the ionophore associated with it is functionally impermeable to divalent cations. DMPP- as well as K(+)-evoked Ca2+ and Mn2+ influx were blocked completely by Ni2+ but only partially by dihydropyridines, suggesting that, in addition to L-type Ca2+ channels, other Ca2+ entry pathways may be present. Inactivation of Ca2+ channels, followed by comparing the rates of Mn2+ uptake at different time periods after the addition of DMPP or high K+, did not happen in the absence of extracellular Ca2+. When 1 mM Ca2+ was present, a delayed inhibition (half time, 10-20 s) was observed, suggesting that it is not due to the entry of Ca2+ itself but to the increase of the cytoplasmic Ca2+ concentration ([Ca2+]i) that takes a few seconds to develop. The influx of Ca2+, estimated from the increase of [Ca2+]i, was also impaired in a time-dependent fashion by previous entry of Mn2+. Inactivation of Ca2+ entry was achieved at estimated mean intracellular Mn2+ concentrations as low as 10(-9) M.

Preconditioning stimuli that augment chromaffin cell secretion

2009 ◽  
Vol 296 (4) ◽  
pp. C792-C800 ◽  
Author(s):  
Laura Tapia ◽  
Josefina García-Eguiagaray ◽  
Antonio G. García ◽  
Luis Gandía
Keyword(s):  
High Frequency ◽  
Chromaffin Cells ◽  
Chromaffin Cell ◽  
Low Frequency ◽  
Half Time ◽  
Adrenal Chromaffin Cells ◽  
Cell Secretion ◽  
Adrenal Chromaffin ◽  
Sympathetic Discharge ◽  
Stimulation Of

We have investigated here whether a preconditioned stimulation of nicotinic and muscarinic receptors augmented the catecholamine release responses elicited by supramaximal 3-s pulses of 100 μM acetylcholine (100ACh) or 100 mM K+ (100K+) applied to fast-perifused bovine adrenal chromaffin cells. Threshold concentrations of nicotine (1–3 μM) that caused only a tiny secretion did, however, augment the responses elicited by 100ACh or 100K+ by 2- to 3.5-fold. This effect was suppressed by mecamylamine and by Ca2+ deprivation, was developed with a half-time ( t1/2) of 1 min, and was reversible. The nicotine effect was mimicked by threshold concentrations of ACh, choline, epibatidine, and oxotremorine-M but not by methacholine. Threshold concentrations of K+ caused lesser potentiation of secretion compared with that of threshold nicotine. The data are compatible with an hypothesis implying 1) that continuous low-frequency sympathetic discharge places chromaffin cells at the adrenal gland in a permanent “hypersensitive” state; and 2) this allows an explosive secretion of catecholamines by high-frequency sympathetic discharge during stress.

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