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Impact of Chromogranin A deficiency on catecholamine storage, catecholamine granule morphology and chromaffin cell energy metabolism in vivo

Abstract

Chromogranin A (CgA) is a prohormone and granulogenic factor in neuroendocrine tissues with a regulated secretory pathway. The impact of CgA depletion on secretory granule formation has been previously demonstrated in cell culture. However, studies linking the structural effects of CgA deficiency with secretory performance and cell metabolism in the adrenomedullary chromaffin cells in vivo have not previously been reported. Adrenomedullary content of the secreted adrenal catecholamines norepinephrine (NE) and epinephrine (EPI) was decreased 30–40 % in Chga-KO mice. Quantification of NE and EPI-storing dense core (DC) vesicles (DCV) revealed decreased DCV numbers in chromaffin cells in Chga-KO mice. For both cell types, the DCV diameter in Chga-KO mice was less (100–200 nm) than in WT mice (200–350 nm). The volume density of the vesicle and vesicle number was also lower in Chga-KO mice. Chga-KO mice showed an ~47 % increase in DCV/DC ratio, implying vesicle swelling due to increased osmotically active free catecholamines. Upon challenge with 2 U/kg insulin, there was a diminution in adrenomedullary EPI, no change in NE and a very large increase in the EPI and NE precursor dopamine (DA), consistent with increased catecholamine biosynthesis during prolonged secretion. We found dilated mitochondrial cristae, endoplasmic reticulum and Golgi complex, as well as increased synaptic mitochondria, synaptic vesicles and glycogen granules in Chga-KO mice compared to WT mice, suggesting that decreased granulogenesis and catecholamine storage in CgA-deficient mouse adrenal medulla is compensated by increased VMAT-dependent catecholamine update into storage vesicles, at the expense of enhanced energy expenditure by the chromaffin cell.

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Abbreviations

CA:

Catecholamines

CgA:

Chromogranin A protein

Chga:

Mouse Chromogranin A gene

DA:

Dopamine

DC:

Dense core

DCV:

DC vesicle

EPI:

Epinephrine

GC:

Golgi complex

KO:

Knockout

NE:

Norepinephrine

SDCV:

Decreased DC with swelled halo

SEV:

Swelled empty vesicle

SV:

Synaptic vesicle

TEM:

Transmission electron microscopy

VMAT:

Vesicular monoamine transporter

WT:

Wild-type

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Acknowledgments

Transmission Electron Microscopy was conducted at the Cellular & Molecular Medicine Electron Microscopy Core Facility at UCSD. A Research Career Scientist Award from the Department of Veterans Affairs supports S.K.M. Mahata’s personal fund and UCSD Academic Senate Grant (RO091B) supported this work.

Conflict of interest

The authors have nothing to declare.

Author information

Correspondence to Sushil K. Mahata.

Additional information

Teresa Pasqua and Sumana Mahata contributed equally to this work.

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Pasqua, T., Mahata, S., Bandyopadhyay, G.K. et al. Impact of Chromogranin A deficiency on catecholamine storage, catecholamine granule morphology and chromaffin cell energy metabolism in vivo. Cell Tissue Res 363, 693–712 (2016). https://doi.org/10.1007/s00441-015-2316-3

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Keywords

  • Norepinephrine
  • Epinephrine
  • Dense core vesicle
  • Synaptic vesicle
  • Mitochondria