Mitochondria are semi-autonomous organelles which are endowed with the ability to change their shape (e.g., by elongation, shortening, branching, buckling, swelling) and their location inside a living cell. In addition they may fuse or divide. These dynamics are discussed. Dislocation of mitochondria may result from their interaction with elements of the cytoskeleton, with microtubules in particular, and from processes intrinsic to the mitochondria themselves. Morphological criteria and differences in the fate of some mitochondria argue for the presence of more than one mitochondrial population in some animal cells. Whether these reflect genetic differences remains obscure. Emphasis is laid on the methods for visualizing mitochondria in cells and following their behaviour. Fluorescence methods provide unique possibilities because of their high resolving power and because some of the mitochondria-specific fluorochromes can be used to reveal the membrane potential. Fusion and fission often occur in short time intervals within the same group of mitochondria. At sites of fusion of two mitochondria material of the inner membrane, the matrix compartment seems to accumulate. The original arrangement of the fusion partners is maintained for some minutes. Fission is a dynamic event which, like fusion, in most cases observed in vertebrate cell cultures is not a straight forward process but rather requires several "trials" until the division finally occurs. Regarding fusion and fission hitherto unpublished phase contrast micrographs, and electron micrographs have been included.

Dynamics of mitochondria in living cells: Shape changes, dislocations, fusion, and fission of mitochondria

Neuronal activity triggers calcium waves in hippocampal astrocyte networks

Swelling of primary astrocyte cultures by exposing them to hypotonic media caused release of label after the cells had been allowed to accumulate 3H-L-glutamate, 3H-D-aspartate, or 3H-taurine. Comparable release of endogenous L-glutamate or taurine, as measured by high- pressure liquid chromatography (HPLC), was also found. Release of label was not affected by treating the cells with cytochalasin B, indicating that microfilament polymerization was not significantly involved. Hypotonic-induced release did not appear to principally involve reversal of the Na(+)-dependent uptake system since increasing external K+ to depolarize the cells by replacement of external Na+, thus maintaining isotonic conditions, increased release to a lesser extent. Threo beta-hydroxyaspartate, a potent 3H-L-glutamate uptake blocker, added externally stimulated efflux of 3H-L-glutamate independently of the swelling-induced efflux. Upon restoration of swollen cells to isotonic medium they showed an unimpaired ability to take up 3H-L- glutamate. The swelling-induced release of label was inhibited by a number of anion transport inhibitors, one of which has been shown to significantly improve outcome in an experimental brain trauma/hypoxia model in which astrocyte swelling is an early event.

https://www.jneurosci.org/content/jneuro/10/5/1583.full.pdf

Swelling-induced release of glutamate, aspartate, and taurine from astrocyte cultures

Role of glial cells in the regulation of the brain ion microenvironment

Astrocytes in culture regulate their volume under anisosmotic conditions by as yet unclear mechanisms. In a number of other cells this process involves a loss of intracellular osmotically active solutes, including taurine. The possibility that taurine participates as an osmoeffector in astrocytes was examined in cultured astrocytes exposed to hyposmolar conditions. Astrocytes responded to decreases in osmolarity by rapid swelling followed by a volume regulatory phase. Hyposmotic conditions induced a dramatic increase of 3H-taurine efflux, with a time course corresponding to the cell volume regulatory phase. Decreasing osmolarity from 310 to 254, 198 or 150 m osmoles resulted in the release of 8.2%, 17%, and 54%, respectively, of 3H-taurine previously accumulated by astrocytes. Endogenous taurine concentration decreased 64%. The efflux of 3-H GABA, 3H-glycine, or 3H-D-aspartate was much less affected under similar conditions. These results suggest a role for taurine as an osmoeffector in astrocytes.

Volume regulation in astrocytes: a role for taurine as an osmoeffector.

Cell volume regulation and energy metabolism were studied in primary cultured cerebral astrocytes during exposure to media of altered osmolarity. Cells suspended in medium containing 1/2 the normal concentration of NaCl (hypoosmotic) swell immediately to a volume 40–50% larger than cells suspended in isoosmotic medium. The cell volume in hypoosmotic medium then decreases over 30 min to a volume approximately 25% larger than cells in isoosmotic medium. In hyperosmotic medium (containing twice the normal concentration of NaCl), astrocytes shrink by 29%. Little volume change occurs following this initial shrinkage. Cells resuspended in isoosmotic medium after a 30 min incubation in hypoosmotic medium shrink immediately to a volume 10% less than the volume of cells incubated continuously in isoosmotic medium. Thus, the regulatory volume decrease (RVD) in hypoosmotic medium involves a net reduction of intracellular osmoles. The RVD is partially blocked by inhibitors of mitochondrial electron transport but is unaffected by an inhibitor of glycolysis or by an uncoupler of oxidative phosphorylation. Inhibition of RVD by these metabolic agents is correlated with decreased cellular ATP levels. Ouabain, added immediately after hypoosmotic induced swelling, completely inhibits RVD, but does not alter cell volume if added after RVD has taken place. Ouabain also inhibits cell respiration 27% more in hypoosmotic medium than in isoosmotic medium indicating that the (Na, K)-ATPase-coupled ion pump is more active in the hypoosmotic medium. These data suggest that the cell volume response of astrocytes in hypoosmotic medium involves the net movement of osmoles by a mechanism dependent on cellular energy and tightly coupled to the (Na, K)-ATPase ion pump. This process may be important in the energy-dependent osmoregulation in the brain, a critical role attributed to the astrocyte in vivo.

Energy-dependent volume regulation in primary cultured cerebral astrocytes.

Astrocytes from primary culture were exposed to conditions that affect intracellular and extracellular Ca2+ concentrations. Astrocyte cell volume was increased approximately 16% after a 30 min exposure to isoosmotic phosphate-buffered saline (PBS) containing the Ca2+ buffer EDTA. Cell volume returned to control values within 30 min of resuspension in normal PBS. Cellular calcium content was not affected by these treatments; however, the recovery of normal cell volume following EDTA exposure was inhibited by 0.1-1.0 mM quinine HCl in a dose-dependent fashion suggesting that a potassium channel controlled by the intracellular Ca2+ concentration is important in this volume response. Intracellular accumulation of an exogenous Ca2+ buffer, BAPTA, also produced cell swelling that persisted following resuspension in normal PBS. Lowering the extracellular Ca2+ concentration with EDTA enhanced the swelling of BAPTA-loaded cells. These data suggest that conditions leading to a decrease in free intracellular Ca2+ concentration may influence astrocyte volume by a mechanism similar to that described in other cell types.

Control of astrocyte volume by intracellular and extracellular Ca2

Secalonic acid D (SAD) is a mycotoxin produced in laboratory cultures by Penicillium oxalicum. Reports have indicated that P. oxalicurn frequently contaminated freshly harvested corn (Mislivec and Tuite, 1970), but none have reported any natural contamination of corn with SAD.

Penicillium Oxalicum and Secalonic Acid D in Fresh Corn

Secalonic acid D (SAD), a hepatotoxic, teratogenic, and slightly mutagenic metabolite of Penicillium oxalicum has been identified as a natural contaminant of grain dust. Secalonic acid D was admini...

Deborah S. Fleischhacker

Papers

Energy-dependent volume regulation in primary cultured cerebral astrocytes.

Control of astrocyte volume by intracellular and extracellular Ca2

Penicillium Oxalicum and Secalonic Acid D in Fresh Corn

Enhancement of Influenza Virus Infections by Secalonic Acid D