The response of yeast to sharp environmental increases in calcium concentration has been extensively studied. However, systematic studies of the response under more general changes are still lacking. Only limited expl...The response of yeast to sharp environmental increases in calcium concentration has been extensively studied. However, systematic studies of the response under more general changes are still lacking. Only limited exploration of cellular responses has been conducted where calcium concentration is decreased. This article describes a set of luminometric experiments that monitor the cytosolic calcium concentration under changing external concentration conditions. As a decrease in external calcium concentrations requires the use of large sample volumes, the experiments require the use of equipment adapted for this purpose. We describe the modification of commercial luminometric equipment to make the exploration possible. We explore the yeast cellular behavior when an increase in external calcium concentration is followed by a decrease in external calcium concentration. We compare these results with those from the case of a double pulse of concentration increase. Results from the experiment show that the first, concentration increasing pulse produces the well-known sharp increase in cytosolic calcium followed by calcium sequestration to return to a cytosolic concentration near its initial condition. Surprisingly, the calcium decrease step shows similar results with a cytosolic increase followed by a return to lower levels. The results suggest the presence of a calcium sensing mechanism regulating calcium influx from external sources. This mechanism would produce channel opening as a response to any changes in external concentration, be it an enhancement or a depletion.展开更多
文摘The response of yeast to sharp environmental increases in calcium concentration has been extensively studied. However, systematic studies of the response under more general changes are still lacking. Only limited exploration of cellular responses has been conducted where calcium concentration is decreased. This article describes a set of luminometric experiments that monitor the cytosolic calcium concentration under changing external concentration conditions. As a decrease in external calcium concentrations requires the use of large sample volumes, the experiments require the use of equipment adapted for this purpose. We describe the modification of commercial luminometric equipment to make the exploration possible. We explore the yeast cellular behavior when an increase in external calcium concentration is followed by a decrease in external calcium concentration. We compare these results with those from the case of a double pulse of concentration increase. Results from the experiment show that the first, concentration increasing pulse produces the well-known sharp increase in cytosolic calcium followed by calcium sequestration to return to a cytosolic concentration near its initial condition. Surprisingly, the calcium decrease step shows similar results with a cytosolic increase followed by a return to lower levels. The results suggest the presence of a calcium sensing mechanism regulating calcium influx from external sources. This mechanism would produce channel opening as a response to any changes in external concentration, be it an enhancement or a depletion.
