Intracellular calcium in mammalian brain cells: fluorescence measurements with quin2

1987 ◽  
Vol 65 (3) ◽  
Author(s):  
M.E. Morris ◽  
J.J. Friedlich ◽  
J.F. MacDonald
Keyword(s):  
Intracellular Calcium ◽  
Mammalian Brain ◽  
Brain Cells ◽  
Fluorescence Measurements

Down-regulation of free intracellular calcium in dissociated brain cells of aged mice and rats

Life Sciences ◽  
1996 ◽  
Vol 59 (5-6) ◽  
pp. 435-449 ◽  
Author(s):  
Henrike Hartmann ◽  
Anne Eckert ◽  
Karsten Velbinger ◽  
Michael Rewsin ◽  
Walter E. Müller
Keyword(s):  
Intracellular Calcium ◽  
Brain Cells ◽  
Down Regulation ◽  
Aged Mice ◽  
Free Intracellular Calcium ◽  
Dissociated Brain Cells

scholarly journals The Chemically Synthesized Ageladine A-Derivative LysoGlow84 Stains Lysosomes in Viable Mammalian Brain Cells and Specific Structures in the Marine Flatworm Macrostomum lignano

Marine Drugs ◽  
2015 ◽  
Vol 13 (2) ◽  
pp. 920-935 ◽  
Author(s):  
Thorsten Mordhorst ◽  
Sushil Awal ◽  
Sebastian Jordan ◽  
Charlotte Petters ◽  
Linda Sartoris ◽  
...  
Keyword(s):  
Mammalian Brain ◽  
Brain Cells ◽  
Macrostomum Lignano

Intracellular calcium recordings from isolated cells of the mammalian central nervous system

1987 ◽  
Vol 65 (5) ◽  
pp. 926-933 ◽  
Author(s):  
M. E. Morris ◽  
J. F. MacDonald ◽  
J. J. Friedlich ◽  
I. Szekelyhidi
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Intracellular Calcium ◽  
Mouse Embryo ◽  
Dorsal Root Ganglion Neurons ◽  
Brain Cells ◽  
Isolated Cells ◽  
Mammalian Central Nervous System ◽  
Intracellular Ca ◽  
Ganglion Neurons

Measurements made with two different techniques of intracellular calcium levels from small isolated cells of the mammalian central nervous system are described and compared. Recordings in cultured mouse embryo spinal cord and dorsal root ganglion neurons, made with double-barrelled borosilicate Ca2+-selective microelectrodes yielded a mean Ca2+ level of 2.3 (SE ± 0.54) μM for the lowest values recorded in 24 out of 46 cells. Intracellular Ca2+ dependence on membrane potential was apparent with levels of calcium ≥4 μM (r = 0.371, n = 29). Both cyclic fluctuations induced by tetraethylammonium and an apparent increase in Ca2+ evoked by the depolarizing excitatory amino acid, L-aspartate, were observed. In contrast, estimates of intracellular Ca2+ obtained by spectrofluorimetry of suspensions of mouse embryo brain cells, loaded with the intracellular Ca-binding fluorescent probe, quin2 provided a [Formula: see text]-fold lower value, 152 (SE ± 7) nM. This more closely resembles levels reported for large neurons where large-tip microelectrodes with greater sensitivity were used, and in spite of the heterogeneity of the cells this value is presumed to be a more accurate estimate of intraneuronal Ca2+ concentration. In these fluorescence studies KCl readily evoked increases in intracellular Ca2+ which could be blocked by verapamil and Cd2+ and were not induced in the absence of Ca2+. Increases were also produced by N-methyl-D-aspartate, but not by the kainate-like Lathyrus neurotoxin, L-3-oxalylamino-2-aminopropionic acid. These results provide preliminary evidence for both voltage-sensitive and receptor-activated Ca channels in embryonic brain cells. Although the recording of intraneuronal Ca2+ with conventional ion-selective microelectrodes in small cells has problems with respect to accuracy, stability, and time constant, recent advances in the design of Ca2+ sensors and electrodes are promising. These, as well as developments in techniques of single cell fluorescence analysis, now offer methods with improved and powerful capacity for accurate and simultaneous measurements of intracellular Ca2+ and membrane electrophysiological parameters.

Arachidonic acid and its metabolites increase Cai in cultured rat oligodendrocytes

1993 ◽  
Vol 264 (3) ◽  
pp. C632-C640 ◽  
Author(s):  
B. Soliven ◽  
M. Takeda ◽  
T. Shandy ◽  
D. J. Nelson
Keyword(s):  
Spinal Cord ◽  
Arachidonic Acid ◽  
Linoleic Acid ◽  
Intracellular Calcium ◽  
Protein Interaction ◽  
Membrane Lipid ◽  
Rat Spinal Cord ◽  
Fura 2 ◽  
Fluorescence Measurements ◽  
Lipid Protein Interaction

Fluorescence measurements of intracellular calcium (Cai) were made on cultured rat spinal cord oligodendrocytes (OLGs) using the dye fura-2. Exposure of OLGs to arachidonic acid (AA) (5-50 microM) elicited a concentration-dependent increase in Cai that was derived mainly from extracellular Ca2+. AA at 50 microM also released Ca2+ from intracellular stores. The response to AA was not decreased by nifedipine or by inhibition of Na(+)-Ca2+ exchange. AA-induced Ca2+ influx pathway was permeable to Mn2+ and Co2+ but not to Ba2+ and was not markedly influenced by depolarization, suggesting that AA activates a voltage-independent, not strictly selective, Ca2+ channel. The Cai response to AA was partially attenuated in the presence of indomethacin, indicating that the Cai response was mediated in part by cyclooxygenase products of AA. However, the AA-induced Cai response far exceeded that induced by prostaglandins and was mimicked by linoleic acid. We conclude that AA modulates Cai of OLGs via two mechanisms: 1) indirectly via cyclooxygenase pathway and 2) directly via membrane lipid-protein interaction.

scholarly journals Dexamethasone stimulates store-operated calcium entry and protein degradation in cultured L6 myotubes through a phospholipase A2-dependent mechanism

2010 ◽  
Vol 298 (5) ◽  
pp. C1127-C1139 ◽  
Author(s):  
Kiyoshi Itagaki ◽  
Michael Menconi ◽  
Bozena Antoniu ◽  
Qin Zhang ◽  
Patricia Gonnella ◽  
...  
Keyword(s):  
Calcium Channel ◽  
Protein Degradation ◽  
Intracellular Calcium ◽  
Muscle Wasting ◽  
Muscle Protein ◽  
Calcium Entry ◽  
Calcium Stores ◽  
Fluorescence Measurements ◽  
Store Operated Calcium Entry

Muscle wasting in various catabolic conditions is at least in part regulated by glucocorticoids. Increased calcium levels have been reported in atrophying muscle. Mechanisms regulating calcium homeostasis in muscle wasting, in particular the role of glucocorticoids, are poorly understood. Here we tested the hypothesis that glucocorticoids increase intracellular calcium concentrations in skeletal muscle and stimulate store-operated calcium entry (SOCE) and that these effects of glucocorticoids may at least in part be responsible for glucocorticoid-induced protein degradation. Treatment of cultured myotubes with dexamethasone, a frequently used in vitro model of muscle wasting, resulted in increased intracellular calcium concentrations determined by fura-2 AM fluorescence measurements. When SOCE was measured by using calcium “add-back” to muscle cells after depletion of intracellular calcium stores, results showed that SOCE was increased 15–25% by dexamethasone and that this response to dexamethasone was inhibited by the store-operated calcium channel blocker BTP2. Dexamethasone treatment stimulated the activity of calcium-independent phospholipase A2(iPLA2), and dexamethasone-induced increase in SOCE was reduced by the iPLA2inhibitor bromoenol lactone (BEL). In additional experiments, treatment of myotubes with the store-operated calcium channel inhibitor gadolinium ion or BEL reduced dexamethasone-induced increase in protein degradation. Taken together, the results suggest that glucocorticoids increase calcium concentrations in myocytes and stimulate iPLA2-dependent SOCE and that glucocorticoid-induced muscle protein degradation may at least in part be regulated by increased iPLA2activity, SOCE, and cellular calcium levels.

scholarly journals Mitotic defects lead to neuronal aneuploidy and apoptosis in frontotemporal lobar degeneration caused by MAPT mutations

2018 ◽  
Vol 29 (5) ◽  
pp. 575-586 ◽  
Author(s):  
Julbert Caneus ◽  
Antoneta Granic ◽  
Rosa Rademakers ◽  
Dennis W. Dickson ◽  
Christina M. Coughlan ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Frontotemporal Lobar Degeneration ◽  
Gene Dosage ◽  
Human Lymphocytes ◽  
Chromosome Instability ◽  
Mammalian Brain ◽  
Brain Cells ◽  
Brain Neurons ◽  
Normal Human ◽  
Mutant Forms

Mutant Tau (MAPT) can lead to frontotemporal lobar degeneration (FTLD). Previous studies associated MAPT mutations and altered function with aneuploidy and chromosome instability in human lymphocytes and in Drosophila development. Here we examine whether FTLD-causing mutations in human MAPT induce aneuploidy and apoptosis in the mammalian brain. First, aneuploidy was found in brain cells from MAPT mutant transgenic mice expressing FTLD mutant human MAPT. Then brain neurons from mice homozygous or heterozygous for the Tau (Mapt) null allele were found to exhibit increasing levels of aneuploidy with decreasing Tau gene dosage. To determine whether aneuploidy leads to neurodegeneration in FTLD, we measured aneuploidy and apoptosis in brain cells from patients with MAPT mutations and identified both increased aneuploidy and apoptosis in the same brain neurons and glia. To determine whether there is a direct relationship between MAPT-induced aneuploidy and apoptosis, we expressed FTLD-causing mutant forms of MAPT in karyotypically normal human cells and found that they cause aneuploidy and mitotic spindle defects that then result in apoptosis. Collectively, our findings reveal a neurodegenerative pathway in FTLD-MAPT in which neurons and glia exhibit mitotic spindle abnormalities, chromosome mis-segregation, and aneuploidy, which then lead to apoptosis.

The differential distribution of beta tubulin mRNAs in individual mammalian brain cells

1985 ◽  
Vol 27 (3) ◽  
pp. 205-214 ◽  
Author(s):  
W. Sue T. Griffin ◽  
Michael A. Alejos ◽  
Erica J. Cox ◽  
Marcelle R. Morrison
Keyword(s):  
Mammalian Brain ◽  
Brain Cells ◽  
Differential Distribution ◽  
Beta Tubulin
Sign in / Sign up

Export Citation Format

Share Document