Insulin, an amino-acid based protein, helps make up a larger polypeptide chain, proinsulin, which is located within the beta cells of the pancreas. Enzymes release the insulin from the larger proinsulin chain just before it is secreted from the beta cells. Insulin’s main function is to lower blood glucose levels by speeding up the membrane transport of glucose from the blood and into the body’s cells, for use for energy or conversion to other storage forms, like glycogen or fats.[i] It also acts as an inhibitor to glycogen by not allowing it to break down into glucose and thus counter acts any metabolic activity that would increase the plasma levels of glucose.[ii]
This experiment will study the effects of insulin on a normal rat, and on a rat that has been induced with Type 1 diabetes. Type 1 diabetes is an autoimmune disorder in which the body attacks the insulin producing beta cells of the pancreas. The pancreas is then incapable of producing insulin, and thus the body’s blood glucose levels become elevated.[iii] Hypothesis
Based on the given information for type 1 diabetes, in this report, a hypothesis can be made that insulin given to a subject with type 1 diabetes would help lower his or her blood glucose levels while insulin that is given to a normal subject will have no effect.
Materials and Methods
• 5 test tubes
• 1 bottle of Glucose Standard
• 1 bottle of deionized water
• 1 bottle of Enzyme Reactor Agent
5 test tubes were filled with glucose standard. The first tube received 1 drop of the glucose standard, the second, 2 drops and so on. Each tube, receiving one drop more than the last. Deionized water was added to tubes one through four, with tube one receiving 4 drops, tube two: three drops, tube three: 2 drops and finally tube four with 1 drop. All five test tubes should now contain the same liquid volume amount. The test tubes were then mixed and placed in the centrifuge. Once extracted from the centrifuge, the pellets were removed from each tube via tweezers. Five drops of the enzyme color regent were added to each tube and then the tubes were incubated. Once removed from the incubator, each tube was individually placed in the spectrophotometer to be analyzed. The results are recorded and graphed to create a standard curve to use as a comparison for the second experiment.
• 2 lab rats
• 4 test tubes
• 6 syringes
• Bottle of Saline
• Bottle of Insulin
• Bottle of Alloxan
• Deinoized Water
• Bottle of Barium Hydroxide
2 lab rats were labeled and separated for this experiment. The first rat was used as the control subject while the second rat was the experimental subject. Rat one was injected with the saline and rat two was injected with the Alloxan. Blood was extracted separately from each rat, via syringe, and placed in the allotted test tube (blood from subject one is labeled R1 and blood from subject two is R2). They were then placed in test tube holder for further use. Rat one and rat two were both injected with insulin. After a period of time, blood samples were again extracted from the rats and placed in separate, labeled test tubes (rat one R1I and rat two, R2I) and placed into the test tube holder. Five drops on deionized water were added to the four test tubes. Following the deionized water, 5 drops of barium hydroxide (used for clearing proteins and cells so a clear glucose reading can be obtained) and finally Heparin (an anticoagulant that stops blood from clotting) were added to the blood samples. The test tubes were then mixed and placed into...