Journal
EUROPEAN JOURNAL OF HISTOCHEMISTRY
Volume 58, Issue 3, Pages 251-257Publisher
PAGEPRESS PUBL
DOI: 10.4081/ejh.2014.2377
Keywords
Brown adipocyte; mitochondrial dynamics; calcium; endoplasmic reticulum
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Funding
- Serbian Ministry of Education, Science and Technological Development [173055, 173054]
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Mitochondria are key organelles maintaining cellular bioenergetics and integrity, and their regulation of [Ca2+](i) homeostasis has been investigated in many cell types. We investigated the short-term Ca-SANDOZ (R) treatment on brown adipocyte mitochondria, using imaging and molecular biology techniques. Two-month-old male Wistar rats were divided into two groups: Ca-SANDOZ (R) drinking or tap water (control) drinking for three days. Alizarin Red S staining showed increased Ca2+ level in the brown adipocytes of treated rats, and potassium pyroantimonate staining localized electron-dense regions in the cytoplasm, mitochondria and around lipid droplets. Ca-SANDOZ (R) decreased mitochondrial number, but increased their size and mitochondrial cristae volume. Transmission electron microscopy revealed numerous enlarged and fusioned-like mitochondria in the Ca-SANDOZ (R) treated group compared to the control, and megamitochondria in some brown adipocytes. The Ca2+ diet affected mitochondrial fusion as mitofusin 1 (MFN1) and mitofusin 2 (MFN2) were increased, and mitochondrial fission as dynamin related protein 1 (DRP1) was decreased. Confocal microscopy showed a higher colocalization rate between functional mitochondria and endoplasmic reticulum (ER). The level of uncoupling protein-1 (UCP1) was elevated, which was confirmed by immunohistochemistry and Western blot analysis. These results suggest that Ca-SANDOZ (R) stimulates mitochondrial fusion, increases mitochondrial-ER contacts and the thermogenic capacity of brown adipocytes.
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