Immunocytochemistry of calciosomes in liver and pancreas

S. Hashimoto, B. Bruno, D. P. Lew, T. Pozzan, P. Volpe, J. Meldolesi

Research output: Contribution to journalArticle

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Abstract

Calciosomes are small cytoplasmic vacuoles identified in various nonmuscle cell types by their content of protein(s) similar to calsequestrin (CS), the Ca2+ storage protein of the muscle sarcoplasmic reticulum (SR). These entities have been interpreted as the 'primitive' counterpart of the SR, and suggested to be the organelle target of inositol-1,4,5-triphosphate action (Volpe, P., K. H. Krause, S. Hashimoto, F. Zorzato, T. Pozzan, J. Meldolesi, and D. P. Lew. Proc. Natl. Acad. Sci. USA. 85: 1091-1095). Immunoperoxidase and immunogold experiments carried out in both thick and ultrathin cryosections of rat hepatocytes and pancreatic acinar cells by using antimuscle CS antibodies revealed a specific labeling widely distributed in the entire cytoplasm, while nuclei were negative. Individual calciosomes appeared as small (105 nm) membrane-bound vacuoles intermingled with, and often apposed to ER cisternae and mitochondria. Other calciosomes were scattered in the Golgi area, in between zymogen granules and beneath the plasma membrane. The cumulative volume of the CS-positive organelles was measured to account for the 0.8 and 0.45% of the cytoplasm in liver and pancreas cells, respectively. The real total volume of the calciosome compartment is expected to be approximately twice as large. In hepatocytes, structures similar to CS-positive calciosomes were decorated by antibodies against the Ca2+ ATPase of muscle SR, while ER cisternae were not. By dual labeling, colocalization was revealed in 53.6% of the organelles, with 37.6% positive for the ATPase only. CS appeared preferentially confined to the content, and the Ca2+ ATPase to the contour of the organelle. The results suggested a partial segregation of the two antigens, reminiscent of their well-known segregation in muscle SR. Additional dual-label experiments demonstrated that hepatic calciosomes express neither two ER markers (cytochrome-P450 and NADH-cytochrome b5 reductase) nor the endolysosome marker, luminal acidity (revealed by 3-[2,4-dinitroanilino)-3'-amino-N-methyl dipropylamine). Calciosomes appear as unique cytological entities, ideally equipped to play a role in the rapid-scale control of the cytosolic-free Ca2+ in nonmuscle cells.

Original languageEnglish
Pages (from-to)2523-2531
Number of pages9
JournalJournal of Cell Biology
Volume107
Issue number6 II
Publication statusPublished - 1988

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Calsequestrin
Sarcoplasmic Reticulum
Pancreas
Organelles
Immunohistochemistry
Liver
Calcium-Transporting ATPases
Vacuoles
Hepatocytes
Cytoplasm
Cytochrome-B(5) Reductase
Muscles
Inositol 1,4,5-Trisphosphate
Muscle Proteins
Antibodies
Acinar Cells
Secretory Vesicles
Cytochrome P-450 Enzyme System
Adenosine Triphosphatases
Mitochondria

ASJC Scopus subject areas

  • Cell Biology

Cite this

Hashimoto, S., Bruno, B., Lew, D. P., Pozzan, T., Volpe, P., & Meldolesi, J. (1988). Immunocytochemistry of calciosomes in liver and pancreas. Journal of Cell Biology, 107(6 II), 2523-2531.

Immunocytochemistry of calciosomes in liver and pancreas. / Hashimoto, S.; Bruno, B.; Lew, D. P.; Pozzan, T.; Volpe, P.; Meldolesi, J.

In: Journal of Cell Biology, Vol. 107, No. 6 II, 1988, p. 2523-2531.

Research output: Contribution to journalArticle

Hashimoto, S, Bruno, B, Lew, DP, Pozzan, T, Volpe, P & Meldolesi, J 1988, 'Immunocytochemistry of calciosomes in liver and pancreas', Journal of Cell Biology, vol. 107, no. 6 II, pp. 2523-2531.
Hashimoto S, Bruno B, Lew DP, Pozzan T, Volpe P, Meldolesi J. Immunocytochemistry of calciosomes in liver and pancreas. Journal of Cell Biology. 1988;107(6 II):2523-2531.
Hashimoto, S. ; Bruno, B. ; Lew, D. P. ; Pozzan, T. ; Volpe, P. ; Meldolesi, J. / Immunocytochemistry of calciosomes in liver and pancreas. In: Journal of Cell Biology. 1988 ; Vol. 107, No. 6 II. pp. 2523-2531.
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abstract = "Calciosomes are small cytoplasmic vacuoles identified in various nonmuscle cell types by their content of protein(s) similar to calsequestrin (CS), the Ca2+ storage protein of the muscle sarcoplasmic reticulum (SR). These entities have been interpreted as the 'primitive' counterpart of the SR, and suggested to be the organelle target of inositol-1,4,5-triphosphate action (Volpe, P., K. H. Krause, S. Hashimoto, F. Zorzato, T. Pozzan, J. Meldolesi, and D. P. Lew. Proc. Natl. Acad. Sci. USA. 85: 1091-1095). Immunoperoxidase and immunogold experiments carried out in both thick and ultrathin cryosections of rat hepatocytes and pancreatic acinar cells by using antimuscle CS antibodies revealed a specific labeling widely distributed in the entire cytoplasm, while nuclei were negative. Individual calciosomes appeared as small (105 nm) membrane-bound vacuoles intermingled with, and often apposed to ER cisternae and mitochondria. Other calciosomes were scattered in the Golgi area, in between zymogen granules and beneath the plasma membrane. The cumulative volume of the CS-positive organelles was measured to account for the 0.8 and 0.45{\%} of the cytoplasm in liver and pancreas cells, respectively. The real total volume of the calciosome compartment is expected to be approximately twice as large. In hepatocytes, structures similar to CS-positive calciosomes were decorated by antibodies against the Ca2+ ATPase of muscle SR, while ER cisternae were not. By dual labeling, colocalization was revealed in 53.6{\%} of the organelles, with 37.6{\%} positive for the ATPase only. CS appeared preferentially confined to the content, and the Ca2+ ATPase to the contour of the organelle. The results suggested a partial segregation of the two antigens, reminiscent of their well-known segregation in muscle SR. Additional dual-label experiments demonstrated that hepatic calciosomes express neither two ER markers (cytochrome-P450 and NADH-cytochrome b5 reductase) nor the endolysosome marker, luminal acidity (revealed by 3-[2,4-dinitroanilino)-3'-amino-N-methyl dipropylamine). Calciosomes appear as unique cytological entities, ideally equipped to play a role in the rapid-scale control of the cytosolic-free Ca2+ in nonmuscle cells.",
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