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PMID: 9285821 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

High-resolution calcium mapping of the endoplasmic reticulum-Golgi-exocytic membrane system. Electron energy loss imaging analysis of quick frozen-freeze dried PC12 cells.

Molecular biology of the cell ·Vol. 8 ·No. 8 ·1997-08-00 ·Pages 1501-12

Pezzati R, Bossi M, Podini P, Meldolesi J, Grohovaz F

Abstract

The calcium pools segregated within the endoplasmic reticulum, Golgi complex, exocytic, and other organelles are believed to participate in the regulation of a variety of cell functions. Until now, however, the precise intracellular distribution of the element had not been established. Here, we report about the first high-resolution calcium mapping obtained in neurosecretory PC12 cells by the imaging mode of the electron energy loss spectroscopy technique. The preparation procedure used included quick freezing of cell monolayers, followed by freeze-drying, fixation with OSO4 vapors, resin embedding, and cutting of very thin sections. Conventional electron microscopy and high-resolution immunocytochemistry revealed a high degree of structural preservation, a condition in which inorganic elements are expected to maintain their native distribution. Within these cells, calcium signals of nucleus, cytosol, and most mitochondria remained below detection, whereas in other organelles specific patterns were identified. In the endoplasmic reticulum, the distribution was heterogeneous with strongly positive cisternae (more often the nuclear envelope and stacks of parallel elements that are frequent in quick frozen preparations) lying in the proximity of or even in direct continuity with other, apparently negative cisternae. The Golgi complexes were labeled strongly and uniformly in all cisternae and part of their vesicles, with no appreciable differences along the cis-trans axis. Weaker or negative signals were recorded from the trans-Golgi network elements and from scattered vesicles, whereas in contrast secretion granules were strongly positive for calcium. These results are discussed in relation to the existing knowledge about the mechanisms of calcium transport in the variations organelles, and about the processes and functions regulated by organelle lumenal calcium in eukaryotic cells.

MeSH Terms
Animals Calcium/metabolism Endoplasmic Reticulum/metabolism,ultrastructure Golgi Apparatus/metabolism,ultrastructure Immunohistochemistry Microscopy, Electron PC12 Cells Rats
Chemicals
Calcium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Pezzati R
Consiglio Nazionale delle Ricerche, Cellular and Molecular Pharmacology Center, Milan, Italy.
Bossi M
Podini P
Meldolesi J
Grohovaz F
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.8.8.1501
Published
1997-08-00
Pages
1501-12
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC276172
Subset
IM
Grants
Telethon · 652 · Italy
External Links
PubMed source DOI: 10.1091/mbc.8.8.1501 PMC Full Text View journal details
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