Developmental regulation of O 2 sensing in neonatal adrenal chromaffin cells from wild-type and NADPH-oxidase-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.

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Development of chromaffin cells depends on MASH1 function

Development ◽

10.1242/dev.129.20.4729 ◽

2002 ◽

Vol 129 (20) ◽

pp. 4729-4738 ◽

Cited By ~ 1

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.

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

Journal of Neurophysiology ◽

10.1152/jn.00669.2020 ◽

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.

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Developmental regulation of leucine-enkephalin expression in adrenal chromaffin cells by glucocorticoids and innervation

Journal of Neuroscience ◽

10.1523/jneurosci.12-10-03818.1992 ◽

1992 ◽

Vol 12 (10) ◽

pp. 3818-3827 ◽

Cited By ~ 14

Author(s):

PD Henion ◽

SC Landis

Keyword(s):

Chromaffin Cells ◽

Developmental Regulation ◽

Adrenal Chromaffin Cells ◽

Leucine Enkephalin ◽

Adrenal Chromaffin

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

AJP Cell Physiology ◽

10.1152/ajpcell.00495.2007 ◽

2008 ◽

Vol 294 (4) ◽

pp. C945-C956 ◽

Cited By ~ 30

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.

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

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100115213 ◽

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.

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