Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.13.12.5 (aequorin)
 1,451 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Aplysia central neurons were injected with the calcium-sensitive photoprotein aequorin and stimulated with trains of identical depolarizing voltage-clamp pulses. The light emissions grew and the outward currents declined in successive pulses. Tetraethylammonium (TEA) enhanced the light emissions to single depolarizing pulses and suppressed the outward current. The remaining net inward current is carried primarily by calcium ions and does not facilitate. The aequorin emissions were larger at all amplitudes of depolarizing pulses that elicited emissions, and the facilitation of emissions in a train of pulses was reduced. The effect of TEA on outward current was nearly maximal when sodium ions were partially replaced with 0.1 M TEA, while the aequorin emissions were further enhanced by increasing the TEA concentration to 0.459 M. TEA enhanced the aequorin emissions at all voltages. These observations suggest that the action of TEA on aequorin emissions is not strictly a consequence of its better known outward current blocking action. The effects of TEA could be partly due to the lowered sodium concentration of these solutions. Replacement of sodium by Tris, sucrose or mannose, however, all produced no enhancement of emissions. Tetramethylammonium (TMA) replacement of sodium had effects similar to those of TEA. Thus TEA and TMA appear to have a specific effect. Part of the enhancement of light emissions by TEA is due to the removal of a series resistance error in the voltage clamp, and this may also account partly for the reduced facilitation of aequorin emissions in TEA. The remainder of the action of TEA on aequorin emissions evidently reflects a specific but previously unrecognized action on the cellular metabolism of calcium ions or on the voltage-dependent calcium channels.
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PMID:Effect of TEA on light emission from aequorin-injected aplysia central neurons. 45 96

1. When molluscan neural somata are filled with the calcium-indicating photo-protein aequorin and subjected to a 1 Hz train of depolarizing pulses (0.3 sec duration to + 15 mV) under voltage clamp, the successive photo-emissions due to calcium influx facilitate. The origin of this phenomenon was investigated in identified neurones from the abdominal ganglion of Aplysia californica.2. Since outward currents inactivate cumulatively in successive pulses, the effective depolarization increases due to a series resistance error. Elimination of this error by electronic compensation or pharmacological block of outward current reduced aequorin response facilitation by only about 30%, on the average.3. When voltage-dependent sodium and potassium currents are blocked in tetraethylammonium (TEA)-substituted zero-sodium sea water, the remaining inward calcium currents display no facilitation. On the contrary, a slow decline during a pulse and a slight progressive depression in successive pulses are observed. Barium-substitution for calcium in the same medium eliminates a small residual potassium current insensitive to external TEA. The remaining inward barium currents also display depression instead of facilitation.4. A non-pharmacological separation of calcium current was accomplished by measuring tail currents at the potassium equilibrium potential following depolarizing pulses. Calcium tail currents activate rapidly and then decline gradually and incompletely as depolarizing pulse duration is lengthened. Tail currents also show no evidence of facilitation; there is instead a slight depression of currents after successive pulses.5. Increments of optical absorbance in neurones filled with the calcium-sensitive dye arsenazo III show a depression rather than facilitation to successive depolarizations in a train. The time course of these absorbance signals is consistent with the time-dependent depression of calcium current.6. Calibration of arsenazo III response amplitude indicates that the dye reports only about 1% of the calcium concentration increment expected from knowledge of cell volume and the charge carried by calcium current during a depolarizing pulse. This suggests that cytoplasmic buffering of free calcium must occur rapidly, on a time scale comparable to the response time of arsenazo III (about 1 msec) or more rapidly.7. The slow potassium tail current following a depolarizing pulse is calcium-dependent and probably provides an approximate index of the internal sub-membrane calcium concentration. Increments in this current after repetitive pulses display a slight progressive depression rather than facilitation.8. Since neither calcium currents nor the concentration transients show facilitation, we conclude that aequorin response facilitation is due to the non-linear dependence of aequorin photo-emissions on calcium concentration. This conclusion is supported by a finding that the very different kinetics of arsenazo III responses and aequorin responses can be reconciled by a simple model representing calcium accumulation and known response properties of the two indicator substances.9. In a train of impulses evoked by injecting depolarizing current into a neurone, the successive action potentials grow in duration. Nevertheless, a nearly constant calcium influx signalled by arsenazo III accompanies broadening action potentials.
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PMID:Aequorin response facilitation and intracellular calcium accumulation in molluscan neurones. 624 86

The effect of raised cytoplasmic pH (pHi) on intracellular concentration ([Ca2+]i) transients following calcium influx during membrane depolarization was studied in identified neurons in the abdominal ganglion of Aplysia californica. The pHi was monitored with pH-sensitive microelectrodes. Sea water containing 15 mM NH4Cl at pH 7.7 elevated pHi about 0.35 pH units from the normal level of 7.17. These cells have an estimated buffering power of about 60 mM/pH unit. Calcium influx was elicited by depolarizing pulses under voltage clamp and [Ca2+]i transients were monitored with the photoprotein aequorin or the metallochromic dye arsenazo III. Aequorin photo-emissions increased by 21--131% (mean, 48%) and arsenazo III absorbance changes accompanying depolarization increased by 9--33% (mean, 20%) after 30 min in NH4+, corresponding roughly to a 14% increase in [Ca2+]i transients. Calcium-dependent potassium tail currents following a depolarizing pulse were somewhat slower and 4--91% (mean, 38%) large in NH4+. The magnitude and time- and voltage-dependence of the membrane calcium conductance was studied using calcium tail currents following depolarizing pulses. The calcium current was unaffected by NH4+, so the enhanced [Ca2+]i transients must reflect reduced calcium buffering at high pHi. Either reduced cytoplasmic calcium binding or slowed active extrusion of calcium may be responsible for this effect.
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PMID:Cytoplasmic alkalization reduces calcium buffering in molluscan central neurons. 627 35

It is shown in this paper that electrical bursting and the oscillations in the intracellular calcium concentration, [Ca2+]i, observed in excitable cells such as pancreatic beta-cells and R-15 cells of the mollusk Aplysia may be driven by a slow oscillation of the calcium concentration in the lumen of the endoplasmic reticulum, [Ca2+]lum. This hypothesis follows from the inclusion of the dynamic changes of [Ca2+]lum in the Chay bursting model. This extended model provides answers to some puzzling phenomena, such as why isolated single pancreatic beta-cells burst with a low frequency while intact beta-cells in an islet burst with a much higher frequency. Verification of the model prediction that [Ca2+]lum is a primary oscillator which drives electrical bursting and [Ca2+]i oscillations in these cells awaits experimental testing. Experiments using fluorescent dyes such as mag-fura-2-AM or aequorin could provide relevant information.
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PMID:Electrical bursting and luminal calcium oscillation in excitable cell models. 898 63