The distribution of mitochondria in pancreatic acinar cells was investigated using confocal fluorescence microscopy and transmission electron microscopy (EM). Acinar cells were studied either after enzymatic isolation or in small segments of undisassociated pancreatic tissue. Loading of isolated acinar cells with Mito Tracker Green or Red, a fluorescence mitochondrial probe, showed that mitochondria are predominantly situated in the perigranular, subplasmalemmal and perinuclear regions. Subsequent applications of EM fixatives induced a leak of the fluorescent indicator to the cytosol but did not change the distribution of mitochondria. EM was then performed on isolated acinar cells and on acinar cells of pancreatic tissue segments. The intracellular distribution of mitochondria was quantified by calculating the percentage of the cross-sectional area that was occupied by mitochondria. In isolated acinar cells the highest density of mitochondria was seen in the perigranular region, where mitochondria occupied 25.69±1.58% of the area, then the subplasmalemmal region with 12.61±0.77% and the perinuclear region with 9.07±0.97% (n=26). Similar results were obtained from acinar cells of pancreatic tissue segments: the perigranular 22.9±1.95%, subplasmalemmal 12.45±0.78% and perinuclear regions 9.07±0.97% (n=26). The outer mitochondrial membranes were frequently positioned close to membranes of the ER, which followed the outer contour of mitochondria. Mitochondria were never found in direct contact with the nuclear envelope: there were usually layers of ER between the mitochondrial and nuclear membranes. Subplasmalemmal mitochondria were found in a very close proximity to the plasma membrane with no ER layers between the mitochondrial and the corresponding plasma membranes. We conclude that in pancreatic acinar cells mitochondria are preferentially distributed to perigranular, subplasmalemmal and perinuclear regions and this distribution is not affected by isolation or fixation procedures.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Bereiter-Hahn J (1990) Behaviour of mitochondria in the living cell. Int Rev Cytol 122:1–63
Bereiter-Hahn J, Voth M (1994) Dynamics of mitochondria in living cells: shape changes, dislocations, fusion and fission of mitochondria. Microsc Res Tech 27:198–219
Buckman JF, Hernandez H, Kress GJ, Voyakova TV, Pal S, Reynolds IJ (2001) MitoTracker labelling in primary neuronal and astrocytic cultures: influence of mitochondrial membrane potential and oxidants. J Neurosci Methods 104:165–176
Collins TJ, Berridge MJ, Lipp P, Bootman MD (2002) Mitochondria are morphologically and functionally heterogeneous within cells. EMBO J 21:1616–1627
Couchman JR, Rees DA (1982) Organelle-cytoskeleton relationships in fibroblasts: mitochondria, Golgi apparatus, and endoplasmic reticulum in phases of movement and growth. Eur J Cell Biol 27:47–54
Csordas G, Thomas AP, Hajnoczky G (1999) Quasi-synaptic Ca2+ signal transmission between endoplasmic reticulum and mitochondria. EMBO J 18:96–108
Dedov VN, Roufogalis BN (1999) Organisation of mitochondria in living sensory neurons. FEBS Letts 456:171–174
Djeza PP, Bortolon R, Perez-Terzic C, Holmuhamedov EL, Terzic A (2002) Energetic communication between mitochondria and nucleus directed by catalyzed phosphotransfer. Proc Natl Acad Sci U S A 99:10156–10161
Duchen MR (1999) Contributions of mitochondria to animal physiology: from homeostatic sensor to calcium signalling and cell death. J Physiol 516:1–17
Frey TG, Mannella CA (2000) The internal structure of mitochondria. Trends Biochem Sci 25:319–324
Gilabert JA, Bakowski D, Parekh AB (2001) Energized mitochondria increase the dynamic range over which inositol 1, 4, 5-trisphosphate activates store-operated calcium influx. EMBO J 20:2672–2679
Glitsch MD, Bakowski D, Parekh AB (2002) Store-operated Ca2+ entry depends on mitochondrial Ca2+ uptake. EMBO J 21:6744–6754
Gonzalez A, Schulz I, Schmid A (2000) Agonist-evoked mitochondrial Ca2+ signals in mouse pancreatic acinar cells. J Biol Chem 275:38680–38686
Hajnoczky G, Robb-Gaspers LD, Seitz MB, Thomas AP (1995) Decoding of cytosolic calcium oscillations in the mitochondria. Cell 82:415–424
Johnson PR, Tepikin AV, Erdemli G (2002) Mitochondria contribute to Ca2+ homeostasis in the mouse pancreatic acinar cell. Cell Calcium 32:56–69
Kern HF (1993) Chapter 2: fine structure of the human exocrine pancreas. In: Vay Liang W, Go et al. (eds) The pancreas: biology, pathology, and disease, 2nd edn. Raven, New York, pp 9–19
Kidd JF, Pilkington MF, Schell MJ, Fogarty KE, Skepper JN, Taylor CW, Thorn P (2002) Paclitaxel affects cytosolic calcium signals by opening the mitochondrial permeability transition pore. J Biol Chem 277:6504–6510
Munn EA (1974) The structure of mitochondria. Academic, New York
Osipchuk YV, Wakui M, Yule DI, Gallacher DV, Petersen OH (1990) Cytoplasmic Ca2+ oscillations evoked by receptor stimulation, G-protein activation, internal application of inositol trisphosphate or Ca2+: simultaneous microfluorimetry and Ca2+-dependent Cl− current recording in single pancreatic acinar cells. EMBO J 9:697–704
Park MK, Ashby MC, Erdemli G, Petersen OH, Tepikin AV (2001) Perinuclear, perigranular and sub-plasmalemmal mitochondria have distinct functions in the regulation of cellular calcium transport. EMBO J 20:1863–1874
Petersen OH (1992) Stimulus-secretion coupling: cytoplasmic calcium signals and the control of ion channels in exocrine acinar cells. J Physiol 448:1–51
Petersen OH, Burdakov D, Tepikin AV (1999) Polarity in intracellular calcium signalling. Bioessays 21:851–860
Pozzan T, Rizzuto R, Volpe P, Meldolesi J (1994) Molecular and cellular physiology of intracellular Ca2+ stores. Physiol Rev 74:595–636
Raraty M, Ward J, Erdemli G, Vaillant C, Neoptolemos JP, Sutton R, Petersen OH (2000) Calcium-dependent enzyme activation and vacuole formation in the apical granular region of pancreatic acinar cells. Proc Natl Acad Sci U S A 97:13126–13131
Rizzuto R, Simpson AWM, Brini M, Pozzan T (1992) Rapid changes of mitochondrial Ca2+ revealed by a specifically targeted recombinant aequorin. Nature 358:325–327
Rizzuto R, Pinton P, Carrington W, Fay FS, Fogarty KE, Lifshitz LS, Tuft RA, Pozzan T (1998) Close contact with endoplasmic reticulum as determinants of mitochondrial Ca2+ responses. Science 280:1763–1766
Scheffler IE (1999) Mitochondria, 1st edn. Wiley, New York
Sjöstrand FS (1976) The problems of preserving molecular structure of cellular components in connection with electron microscopic analysis. J Ultrastruct Res 55:271–280
Sjöstrand FS (1978) The structure of mitochondrial membranes: a new concept. J Ultrastruct Res 64:217–245
Skulachev VP (2001) Mitochondrial filaments and clusters as power-transmitting cables. Trends Biochem Sci 26:23–29
Szalai G, Csordas G, Hantash BM, Thomas AP, Hajnoczky G (2000) Ca2+ signal transmission between ryanodine receptors and mitochondria. J Biol Chem:275 15305–15313
Tinel H, Cancela JM, Mogami H, Gerasimenko JV, Gerasimenko OV, Tepikin AV, Petersen OH (1999) Active mitochondria surrounding the pancreatic acinar granule region prevent spreading of inositol trisphosphate-evoked local cytosolic Ca2+ signals. EMBO J 18:4999–5008
Voronina S, Sukhomlin T, Johnson PR, Erdemli G, Petersen OH, Tepikin AV (2002) Correlation of NADH and Ca2+ signals in mouse pancreatic acinar cells. J Physiol 539.1:41–52
P.R. Johnson and N.J. Dolman contributed equally to the study. This work was supported by a Medical Research Council programme grant. P.R.J. is a Medical Research Council PhD student and N.J.D. is a Wellcome Trust PhD student.
About this article
Cite this article
Johnson, P.R., Dolman, N.J., Pope, M. et al. Non-uniform distribution of mitochondria in pancreatic acinar cells. Cell Tissue Res 313, 37–45 (2003). https://doi.org/10.1007/s00441-003-0741-1
- Electron microscopy
- Mito Tracker Red
- Mouse (CD-1)