The ceramide-enriched <Emphasis Type="Italic">trans</Emphasis>-Golgi compartments reorganize together with other parts of the Golgi apparatus in response to ATP-depletion |
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Authors: | Claudia Meisslitzer-Ruppitsch Clemens Röhrl Carmen Ranftler Josef Neumüller Monika Vetterlein Adolf Ellinger Margit Pavelka |
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Institution: | 1.Department of Cell Biology and Ultrastructure Research, Center for Anatomy and Cell Biology,Medical University of Vienna,Vienna,Austria;2.Division for Pathobiochemistry, Center for Pathobiochemistry and Genetics,Medical University of Vienna,Vienna,Austria |
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Abstract: | In this study, the ceramide-enriched trans-Golgi compartments representing sites of synthesis of sphingomyelin and higher organized lipids were visualized in control
and ATP-depleted hepatoma and endothelial cells using internalization of BODIPY-ceramide and the diaminobenzidine photooxidation
method for combined light-electron microscopical exploration. Metabolic stress induced by lowering the cellular ATP-levels
leads to reorganizations of the Golgi apparatus and the appearance of tubulo-glomerular bodies and networks. The results obtained
with three different protocols, in which BODIPY-ceramide either was applied prior to, concomitantly with, or after ATP-depletion,
revealed that the ceramide-enriched compartments reorganize together with other parts of the Golgi apparatus under these conditions.
They were found closely associated with and integrated in the tubulo-glomerular bodies formed in response to ATP-depletion.
This is in line with the changes of the staining patterns obtained with the Helix pomatia lectin and the GM130 and TGN46 immuno-reactions occurring in response to ATP-depletion and is confirmed by 3D electron tomography.
The 3D reconstructions underlined the glomerular character of the reorganized Golgi apparatus and demonstrated continuities
of ceramide positive and negative parts. Most interestingly, BODIPY-ceramide becomes concentrated in compartments of the tubulo-glomerular
Golgi bodies, even though the reorganization took place before BODIPY-ceramide administration. This indicates maintained functionalities
although the regular Golgi stack organization is abolished; the results provide novel insights into Golgi structure–function
relationships, which might be relevant for cells affected by metabolic stress. |
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