Correlated measurements of free and total intracellular calcium concentration in central nervous system neurons

Pozzo-Miller, Lucas; Pivovarova, N. B.; Connor, John A.; Reese, Thomas S.; Andrews, S. Brian

doi:10.1002/(sici)1097-0029(19990915)46:6<370::aid-jemt5>3.0.co;2-3

Cited by 22 publications

(7 citation statements)

References 71 publications

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“…Baseline levels of the somatic apparent free [Ca 2+ ] i in NR1 R/− cells were similar with those in wild‐type neurons (R/−, 84±20 nM, n =21; +/+, 64±8 nM, n =29; mean±s.e.m., n cells ; Figure 1C), and in line with (Garaschuk et al , 1997; Pozzo‐Miller et al , 1999; Shuttleworth and Connor, 2001). NMDAR‐evoked responses were also comparable between the two genotypes (apparent [Ca 2+ ] i after stimulation: +/+, 631±75 nM, n =29; R/−, 629±86 nM, n =21; for derivation of [Ca 2+ ] i values, see Supplementary data and Supplementary Methods).…”

Section: Resultssupporting

confidence: 75%

CaMKII translocation requires local NMDA receptor-mediated Ca2+ signaling

Thalhammer

Rudhard

Tigaret

et al. 2006

EMBO J

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Excitatory synaptic transmission and plasticity are critically modulated by N‐methyl‐D‐aspartate receptors (NMDARs). Activation of NMDARs elevates intracellular Ca2+ affecting several downstream signaling pathways that involve Ca2+/calmodulin‐dependent protein kinase II (CaMKII). Importantly, NMDAR activation triggers CaMKII translocation to synaptic sites. NMDAR activation failed to induce Ca2+ responses in hippocampal neurons lacking the mandatory NMDAR subunit NR1, and no EGFP‐CaMKIIα translocation was observed. In cells solely expressing Ca2+‐impermeable NMDARs containing NR1N598R‐mutant subunits, prolonged NMDA application elevated internal Ca2+ to the same degree as in wild‐type controls, yet failed to translocate CaMKIIα. Brief local NMDA application evoked smaller Ca2+ transients in dendritic spines of mutant compared to wild‐type cells. CaMKIIα mutants that increase binding to synaptic sites, namely CaMKII‐T286D and CaMKII‐TT305/306VA, rescued the translocation in NR1N598R cells in a glutamate receptor‐subtype‐specific manner. We conclude that CaMKII translocation requires Ca2+ entry directly through NMDARs, rather than other Ca2+ sources activated by NMDARs. Together with the requirement for activated, possibly ligand‐bound, NMDARs as CaMKII binding partners, this suggests that synaptic CaMKII accumulation is an input‐specific signaling event.

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Section: Resultssupporting

confidence: 75%

CaMKII translocation requires local NMDA receptor-mediated Ca2+ signaling

Thalhammer

Rudhard

Tigaret

et al. 2006

EMBO J

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“…3), and ER Ca sequestration is reversible (Pozzo‐Miller et al 1997). The time course of [Ca] ER recovery is surprisingly long, in the order of 15–30 min (Pozzo‐Miller et al 1999). Since ER stores will presumably be loaded and primed for Ca 2+ ‐ or ligand‐gated release during this recovery phase (Figs 1 and 2; Pozzo‐Miller et al 1996), it is likely that ER cisterns will carry a memory trace of previous synaptic activity over the time frame of minutes.…”

Section: Synaptic Activation Of Calcium Influx Induces ‘Superbufferinmentioning

confidence: 99%

“…We review here our studies on the calcium sequestering and release activity of dendritic ER in CA3 hippocampal pyramidal neurons both at rest and at defined times following neuronal activity, using microfluorometric [Ca 2+ ] i imaging (Connor, 1986) and energy‐dispersive X‐ray (EDX) microanalysis (Leapman & Andrews, 1991; Buchanan et al 1993; Andrews et al 1994). The first technique reveals the quantitative distribution of free Ca 2+ in living dendrites, while the latter method provides direct, quantitative, and complementary measurements of total (free plus bound) Ca concentrations within identified subcellular compartments at the electron microscopic level (reviewed by Pozzo‐Miller et al 1999).…”

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confidence: 99%

Microheterogeneity of calcium signalling in dendrites

Pozzo-Miller

Connor

Andrews

2000

The Journal of Physiology

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Transient changes in the intracellular concentration of free Ca2+ ([Ca2+]i) originating from voltage‐ or ligand‐gated influx and by ligand‐ or Ca2+‐gated release from intracellular stores, trigger or modulate many fundamental neuronal processes, including neurotransmitter release and synaptic plasticity. Of the intracellular compartments involved in Ca2+ clearance, the endoplasmic reticulum (ER) has received the most attention because it expresses Ca2+ pumps and Ca2+ channels, thus endowing it with the potential to act as both an intracellular calcium sink and store. We review here our ongoing work on the role of calcium sequestration into, and release from, ER cisterns and the role that this plays in the generation and termination of free [Ca2+]i transients in dendrites of pyramidal neurons in hippocampal slices during and after synaptic activity. These studies have been approached by combining parallel microfluorometric measurements of free cytosolic [Ca2+]i transients with energy‐dispersive X‐ray microanalytical measurements of total Ca content within specific dendritic compartments at the electron microscopy level. Our observations support the emerging realization that specific subsets of dendritic ER cisterns provide spatial and temporal microheterogeneity of Ca2+ signalling, acting not only as a major intracellular Ca sink involved in active clearance mechanisms after voltage‐ and ligand‐gated Ca2+ influx, but also as an intracellular Ca2+ source that can be mobilized by a signal cascade originating at activated synapses.

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“…Such work included electrically excitable cells, such as neuronal cell frag ments (Pozzo-Miller et al, 1999Tareilus et al , 1993) and intact neuronal cells (Heuser et al. 1979: Torri-Tarelli et al.. 1985).…”

Section: A Appreciation Of Methods Availablementioning

confidence: 99%

Sub-Second Cellular Dynamics: Time-Resolved Electron Microscopy and Functional Correlation

Plattner¹,

Hentschel²

2006

International Review of Cytology

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Subcellular processes, from molecular events to organellar responses and cell movement, cover a broad scale in time and space. Clearly the extremes, such as ion channel activation are accessible only by electrophysiology, whereas numerous routine methods exist for relatively slow processes. However, many other processes, from a millisecond time scale on, can be "caught" only by methods providing appropriate time resolution. Fast freezing (cryofixation) is the method of choice in that case. In combination with follow-up methodologies appropriate for electron microscopic (EM) analysis, with all its variations, such technologies can also provide high spatial resolution. Such analyses may include, for example, freeze-fracturing for analyzino restructuring of membrane components, scanning EM and other standard EM techniques, as well as analytical EM analyses. The latter encompass energy-dispersive x-ray microanalysis and electron spectroscopic imaging, all applicable, for instance, to the second messenger, calcium. Most importantly, when conducted in parallel, such analyses can provide a structural background to the functional analyses, such as cyclic nucleotide formation or protein de-or rephosphorylation during cell stimulation. In sum, we discuss many examples of how it is practically possible to achieve strict function-structure correlations in the sub-second time range. We complement this review by discussing alternative methods currently available to analyze fast cellular phenomena occurring in the sub-second time range.

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Correlated measurements of free and total intracellular calcium concentration in central nervous system neurons

Cited by 22 publications

References 71 publications

CaMKII translocation requires local NMDA receptor-mediated Ca2+ signaling

CaMKII translocation requires local NMDA receptor-mediated Ca2+ signaling

Microheterogeneity of calcium signalling in dendrites

Sub-Second Cellular Dynamics: Time-Resolved Electron Microscopy and Functional Correlation

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