The Effect of Glucose and Caffeine on Memory and Learning Time in Mice

Shaina Danet

 

Introduction

            For humans, activities require active use of our brain to decode information and store it into our long term memory. One place where the brain stores the information it acquires is the hippocampus, particularly spatial recognition information (Martinez et al. 1982).Slightly controversial studies have indicated the hippocampus is associated with recognition, but familiarity is distributed to the other cortical areas (Fortin et al 2004). The brain uses glucose as a food source to be able to run (Booth 1994 as described by Donohoe and Benton 1999). Glucose is taken from the foods we eat during digestion, and is released into the blood stream. Other substances, such as caffeine, have also been shown to improve memory through increased alertness (Addicott and Laurienti 2009). Caffeine also shows evidence of increasing brain activity in general, which decreases the time needed for the hippocampus to take in information and create memories.

            In order for the brain to function properly, there has to be a minimum amount of glucose in the blood that can pass the blood-brain barrier and feed the brain (Foster et al 1998). Evidence has suggested that one reason people have cognitive impairments is from having consistent low blood glucose levels (Foster et al 1998). The elderly particularly have this problem because of poor glucose regulation; as the human body ages, the ability to metabolize glucose diminishes, including having drastically reduced endocrine response to stress (Gold 1986, 1992 as said by Foster et al 1998). When giving the elderly regular doses of glucose, memory drastically improves (manning et al 1992, as said by Foster et al. 1998)

            In rodents and humans alike, there has been evidence that glucose administration can enhance memory (Gold 1986, 1991, 1992, as described by Donohoe and Benton 1999; Foster et al 1998; Canal et al 2005) as a result of excess blood glucose, as well as individual differences in glucose regulation (Gonder-Frederick et al 1987, Hall et al 1989, Benton and Owens 1993, and Benton et al 1994, as described by Donohoe and Benton 1999). Glucose has been shown to have positive effects on healthy adults, and improving cognitive function in adults with memory problems (Foster et al 1998). While most information on the effects of glucose on the hippocampus has focused on memory tasks, there has also been evidence that glucose can contribute to better scores on some non-memory tasks (Donohoe and Benton 1999). Owen et al (2010) found that higher dosages (60g compared to 20g) of glucose showed more of an improvement with word recall and delay than the lower dosage. This went against previous research, which found little difference between two dosage levels, though they were closer in amount (2010). Kanoski and Davidson (2010) found that high energy diets can actually impair cognitive function and memory. Therefore, previous data suggests that there is a limit to what a human’s body can handle when it comes to energy being consumed and how much glucose is being released into our blood streams.

            An outside chemical that can have a beneficial effect on memory conversion is caffeine. Caffeine has been shown to enhance memory despite different processes of ingestion (acute, slow delivery, long intermittent consumption) (Cunha and Agostinho 2010). However, other research directly counters the supposed effects of caffeine (Childs and deWit 2006, as told by Cunha and Agostinho 2010). Caffeine is believed to aid memory processing by producing a neurostimulant that provokes adenosine and allows the neurons to be disinhibited (Nehlig 1999 and Ferre 2008, as told by Addicott and Laurienti 2009) as well as increasing cerebral energy metabolism and cortical activity (Arendash et al 2009).The drawback of constant exposure to caffeine is that it reduces the effects that it would have compared to an acute dose (Addicott and Laurienti 2009). There also has been evidence that chronic exposure of caffeine will affect the A1-CB1 receptors in the hippocampus, altering how the brain translates spatial memory (Souse et al 2011). Along those same lines, Arendash et al found that mice who had exhibited the signs of alzheimers actually stopped and slightly reversed memory impairment with a moderate dose of caffeine daily (2009).

                        While previous research has independently assessed the role of glucose and caffeine on memory development, there has yet to be an experiment simultaneously comparing the separate effects of these two compounds. I examined how variable concentrations of glucose and caffeine affect memory in mice using a maze. I had predicted that higher concentrations of glucose and caffeine will have increased memory retention. In addition, I had predicted that glucose will have a greater effect on memory and learning time compared to caffeine.

 

Methods

Preparations:

            Forty mice (Mus musculus) were housed two to a bedded cage, separated by sex, at room temperature with food pellets and water available in the animal room at McKendree University. The mice were kept under a twelve hour light period where the lights were constantly on. Each treatment had cages of eight mice, four females and four males, and the cages for each treatment were grouped together. The five solution treatments that were used to study the effects of glucose and caffeine on memory and learning time were 9% saline solution for control, 2mg/kg glucose, 4mg/kg glucose, 2mg/kg caffeine, and 4mg/kg caffeine. The saline solution was made in a 9% sodium chloride in DI water solution. The four other solutions were made with saline solution and either crystallized glucose or caffeine powder mixed in.  Each mouse was weighed to the nearest gram, then the weights were used to calculate the 2mg substance/kg weight of mouse or 4mg/kg dose that each mouse would receive per 1mL shot. This information was used to create the stock solutions that were kept in 1L bottles. 15mL at a time were portioned into 30mL bottles for easier handling

Trials:

Between October 8th-24th, and February 4th- March 1st I ran trials every Monday, Wednesday and Friday for a total of nine trials. The trials were always started at the same hour. The day before the maze test the mice had their food taken away for fasting [18-24 hours before the trial]. During the testing, the two mice from each cage were moved to a larger holding area where they were given 1ml shots of their respective solution to the base of their tail. After waiting about four minutes for the solution to move into the blood stream, one mouse at a time would be placed at the start of the maze and timed from when they touched the start of the maze until they approached the end of the maze. The food pellets, as well as treats such as chocolate and peanut butter, were placed at the finish line as incentive for the mice to move to the end of the maze. When the mouse approached the food at the finish line the mouse was placed back in their cage with a handful of food. After five trials the maze was reversed for another four trials. This was to keep the data consistent with creating new memories of the maze, instead of timing memory recall. The times for each mouse were averaged based on the maze used and used in the ANOVA. An average of all the mice for each treatment was then done to see if there was a pattern across variables for each maze. The time versus variable data was run with a one-way ANOVA to examine across glucose and caffeine times, as well as the 2mg and 4mg concentrations. The data collected was put into the one-way ANOVA in three groups: Average 1, the variable, and average 2.The ANOVA was run twice, either with average 1 and the variable, or average 2 and the variable, the variable being the dependent factor. The post-hoc test used was the Tukey test.  

Results

Figure 1 The average times across treatments for the mice to complete Maze 1; the 2mg caffeine had the shortest average time. (p=.137)

This shows the comparison of the average times of each treatment. The control group had an average of 1.96 minutes to run the maze. The 2mg Glucose and 4mg Caffeine treatments had a higher average time than the control(2.35 and 2.17 minutes), but the 4mg Glucose and 2mg Caffeine groups had a shorter time(1.81 and 1.68 minutes), with the 2mg Caffeine group having the shortest average time on maze one. The results were not significant between the groups, however (p= .137).

Figure 2 The average times across treatments for the mice to run Maze2 ; 2mg Glucose had the shortest average time, and only 2mg caffeine as at/above the control time. (p=.485)

            The averages for the second maze also had unexpected results. Compared to the control (1.08 minutes), 2mg Caffeine had the same average time. 2mg Glucose had the shortest time with .67 minutes, and then 4mg Glucose with .93 minutes, and finally 4mg Caffeine close behind with .95 minutes. There was also no significance between the groups (p=.485). The level of standard deviation is much greater in figure 2 than figure 1 as well.

 

Discussion

Based on previous research, one would expect glucose and caffeine to have better results than the control. The results for this current experiment came to be vastly different from the literature used. There was no significance between any of the groups, and even in figures 1 and 2 one can see that certain treatments actually had a higher time average than the control. Previous work confirmed that glucose helps increase memory for certain tests, and could even reverse poor cognitive function (Foster et al 1998, Canal et al 2005, Donohoes and Benton 1999, Owen et al 2010). Owen et al found that a higher concentration of glucose (60g) showed significant results compared with placebo, while a lower concentration (25g) did not. In the same though process, caffeine has previously been shown to increase memory retention as well as small improvements towards cognitive deficiencies (Addicott and Laurienti 2009, Cunha and Agostinho 2010, Arendash et al 2009).

            Previous research has indicated that chemicals such as glucose and caffeine can help improve memory and even alleviate certain cognitive disorders. When the brain is transforming information into memory, the brain takes in large amount of glucose to fuel it. If one takes in large amounts of glucose, the brain will have plenty of energy to feed off of, which is supported by previous research (Foster et al 1998, Canal et al 2005, Owen et al 2010). As for caffeine, the stimulant capabilities of caffeine will increase alertness, meaning that the brain is paying special attention to the information flowing in from the senses. When we pay more attention to our surroundings, we are able to help our memory retention, as compared to when a person is fatigued. The fact even chronic caffeine drinkers show memory retention improvement, as compared to the vast increase for acute drinkers, suggests that this increased awareness really does make the difference in memory retention (Addicott and Laurienti 2009). Both glucose and caffeine have also shown evidence of improving and even reversing cognitive defects (Cunha and Agostinho 2010, Arendash et al 2009, Kanoski and Davidson 2010).

            My results are contrary to the results given by my sources. First, there is no descending pattern of averages between the control and the other treatments as would be expected if glucose and caffeine improved memory retention(fig.1 and fig 2). In fact, some of the groups had a longer time average than the control. Also, if higher concentrations of the two solutions really do improve memory retention better than the lesser concentrations, then there would be a trend that the 4mg doses would be smaller than the 2mg doses. In reality there is a mixture between which dosage is shorter between glucose and caffeine, and between the two mazes. Additionally, we find that the results are not significant. This information rejects my original hypothesis that glucose and caffeine have a significant effect on memory retention.

            My results would show that glucose and caffeine does not have much of an effect on the brain, not readily increasing memory retention. This could be just an unusual result, which does happen, or could possibly provide clues towards the specificity of how these two substances help memory. Individuals may be affected differently by different substances, but since this experiment has a large enough testing group that can be ruled out. Another possibility is that the maze testing with a goal for food may not be sufficient for testing the full capability of glucose or caffeine on memory. My research can be used, along with other research that showed no significant results, to determine if there are certain processes within the body that can use external substances to enhance function, or dispose of it normally through waste. There may be a biological process that determines whether the brain will be enhanced with extraneous glucose or caffeine.

            New experiments should test whether other memory retention test provide a better result towards memory retention testing. Some memory tests are simply unaffected by glucose and caffeine additives. Also, one could see if there is a difference between injecting the mouse with the solution vesus having them ingest the glucose or caffeine with normal tap water.

Lit Cited

Addicott, M.A., Laurienti,P.J. 2009. A comparison of the effects of caffeine following abstinence and normal caffeine use. Psychopharmacology. 207:423-431.

Arendash, G.W., Mori, T., Cao, C., Mamcarz, M., Runfeldt, M., Dickson, A., Rezai-Zadeh, K., Tan, J., Citron, B.A., Lin, X., Echeverria, V., Potter, H. 2009. Caffeine reverses cognitive impairment and decreases brain amyloid-ß levels in aged alzheimer’s disease mice. Journal of Alzheimer’s Disease. 17: 661-680.

Benton, D., Owens, D.D. 1993. Blood glucose and human memory. Psychoparmacology. 113: 83-88.

Benton, D., Owens, D.S., Parker, P. Y. 1994. Blood glucose influences memory and attention in younger adults. Neuropsychologia. 32: 595-607.

Booth, D. 1994. The psychology of nutrition. Taylor and Francis, London.

Canal, C.E., Stutz, S.J., Gold, P.E. 2005. Glucose injections into the dorsal hippocampus or dorsolateral striatum of rats prior to T-maze training: Modulation of learning rates and  strategy selection. Learn, Mem, 12: 367-374.

Childs, E., deWit, H. 2006. Subjective, behavioral, and physiological effects of acute caffeine in light, nondependent caffeine users. Psychopharmacology. 185: 514-523

Cunha, R.A., Agostinho, P.M. 2010. Chronic caffeine consumption prevents memory disturbance in different animal models of memory decline. Journal of Alzheimer’s Disease. 20: 95-116.

Donohoe, R.T., Benton, D. 1999. Cognitive functioning is susceptible to the level of blood glucose. Psychopharmacology (Berl). 145: 378-85.

Fernandez, G., Effern, A., Grunwald, T., Pezer, N., Lehnertz, k. 1999. Real-time tracking of memory formation in the human rhinal cortex and hippocampus. Science. 285: 1582-5. 

Ferre, S. 2008. An update on the mechanisms of the psychostimulant effects of caffeine. Journal of Neurochemistry. 105: 1067-1079

Fortin, N.J., Wright, S.P., Eichenbaum, H. 2004. Recollection-like memory retrieval in rats is dependent on the hippocampus. Nature. 431: 188-91.

Foster, J.K., Lidder, P.G., Sunram, S.I. 1998. Glucose and memory: fractionation of  enhancement effects? Psychopharmacology (Berl). 137: 259-70.

Gold, P.E. 1986. Glucose modulation of memory storage processing. Behavioral Neural Biology. 45: 342-349

Gold, P.E. 1991. An integrated memory regulation system: from blood to brain. In: Fredrickson  RCA, McGaugh JL, Felton DL (eds) Peripheral signaling of the brain: role in neural-immune interactions, learning and memory. Hogrefe and Huber, Toronto, pp 391-414.

Gold, P.E. 1992. Modulation of memory processing: enhancement of memory in rodents and humans. In: Squire LR, Buttes N (eds) Neuropsychology of memory. Guildford press, New York, pp 402-414.

Gonder- Frederick, L., Hall, J.L., Vogt, J., Cox, D.J., Green, J., Gold, P.E. 1987. Memory enhancement in elderly humans: effects of glucose ingestion. Physiological Behavior. 41: 503-504

Hall, J.L., Gonder-Frederick, L.A., Chewning W.W., Silveria, J., Gold, P.E. 1989. Glucose enhancement of performance on memory tests in young and aged humans.    Neuropsychologia. 27: 1129-1138.

Kanoski, S.E., Davidson, T.L. 2010. Different patterns of memory impairments accompany          short- and longer-term maintenance on a high-energy diet. Journal of experimental psychology. Animal behavior processes. 36: 313-319.

Manning C.A., Parsons M.W., Gold, P.E. 1992. Anterograde and retrograde enhancement of 24-h memory by glucose in elderly humans. Behavioral Neural Biology. 58:125-130

Martinez , J.L., Petty, C., Messing, R.B. 1982. Regional brain uptake of 2-deoxy-D-glucose  following training in a discriminated y-maze avoidance task. Journal of comparative and  physiological psychology. 96: 721-4.

Nehlig, A. 1999. Are we dependent upon coffee and caffeine? A review on human and animal data. Neuroscience Biobehavior Review. 23:563–576.

Owens, L., Finnegan, Y., Hu, H., Scholey, A.B., Sünram-Lea, S.I. 2010. Glucose effects on long-term memory performance: duration and domain specificity. Psychopharmacology.  211: 131-140.

Sousa, V.C., Assaife-Lopes, N., Ribeiro, J.A., Pratt,J.A., Brett, R.R., Sebastião, A.M. 2011.  Regulation of hippocampal cannabinoid CB1 receptor actions by adenosine A1 receptors  and chronic caffeine administration: implications for the effects of Δ9- tetrahydrocannabinol on spatial memory. Neuropsychopharmacology. 36:472-487.