Caffeine as a pathway for increased Calcium

This experiment determines that caffeine is a pathway for increased Calcium.  Ca2+ is increased by caffeine from 17-30 minutes into the experiment due to the release of the dantrolene (Ca2+ inhibitor), which was present in the beginning.  Fura 2 digital epiflourescence microscopy and Cell Cultures were used to determine the calcium levels. 

Two experiments were compiled in this graph to show that both caffeine and AICAR can increase GLUT 4 production in the cell as compared to a control (the cell under normal circumstances).  The AICAR and caffeine produced more GLUT 4 than the control group and the groups that were given various inhibitors, which effectively block the effects on GLUT 4.  Incubations, Western Blot Analysis, and Cell Cultures were used to gather the information used in this figure. 

2-Deoxyglucose was measured in this experiment in place of glucose, because it can be used in the cell by the same pathway, but is not metabolized in all other cell functions, and is therefore a more accurate representation of the glucose that would be used in this particular pathway.  The experiment shows that insulin increases glucose transport.  The increases due to insulin are amplified in the samples treated with caffeine and AICAR.  Incubations, and Cell Cultures were used to gather the information used in this figure.  This could possibly suggest this as a method of overcoming insulin resistance by increasing GLUT 4 production, which increases the amount of glucose that can enter the cell by way of insulin. 

No increases were observed in GLUT 1 and GLUT 3 due to caffeine or AICAR.  This means that GLUT 4 is specifically increased by these mechanisms.  Incubations, Western Blot Analysis, and Cell Cultures were used to gather the information used in this figure. 

Caffeine produced larger increases in the transcription factors that make GLUT 4 than the control, and the samples treated with both caffeine and an inhibitor.  Western Blot Analysis and Cell Cultures were used to determine this data. 

The GLUT 4 transcription factors were increased with the addition of AICAR as compared to the control group.  Western Blot Analysis and Cell Cultures were used to determine this data. 

Caffeine and Ionomycin (a calcium ionophore, which allows increased calcium levels by diffusion into the cell) produced increases in GLUT 4 and its transcription factors as compared to the control.  Western Blot Analysis and Cell Cultures were used to determine protein levels in this experiment.