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Acute vs Chronic Ingestion of Grape Seed Extract

on the Spatial Memory of Mice


Anna Hedges & Staci Hammer
Loras College
Department of Biology
1450 Alta Vista St.
Dubuque, IA 52001

Statement of Objectives:
The purpose of this experiment is to compare the memory of mice after receiving either chronic
or acute treatments of grape seed extract to see if one type of dosage improves memory. Both
types of doses have been shown to enhance memory by observing faster water maze trial times in
mice over mice who have not received treatment of the grape seed extract. However,
administering both chronic and acute doses have never been implemented in the same
experiment. For this experimental design, there will be three different groups of mice. Over five
days, one group will receive no extract to serve as a control, one group of mice with chronic
treatment will be administered 100 micrograms/g/day of grape seed extract, and the acute group
will get a one-time dose of 500 micrograms/g of the extract. Memory improvement will be
measured by the difference in maze trial times of pre-and post-tests taken before and after all
dosages have been administered. Mice with the prolonged treatment of grape seed extract are
expected to experience a significantly larger decrease in time to complete the maze because the
grape seed extract will have more time to imprint the route into each mouse’s memory.
Project Overview:
Grapes are a type of fruit that contain flavonols, phytochemical compounds that aid in memory
and neuronal activity. Consumption of these assists in memory improvement because flavonols
protect neurons from damage and activity loss (Praag et al., 2007). The protected neurons now
have the ability to build stronger neuronal pathways and connections to retain memory (Subash
et al., 2014). Both acute (Jamshidzadeh, et. al., 2010) and chronic (Sarkaki, et. al., 2013)
treatment have been proven to work. Whereas both long-term and short-term treatments are
effective in reducing the time needed to complete maze trials, the amount or duration of intake
for optimal results remains unclear. Knowing whether grape seed extract improves memory more
by ingestion over time, or in one sitting, can lead to lifestyle changes for those who wish to
maintain or increase their memory capabilities. These flavonols must be digested and sent to the
brain to begin to work on the neurons (Wang et al., 2008). This experiment intends to test the
hypothesis that not only does grape seed extract work to enhance memory, but also the chronic
treatment of grape seed extract will have larger decrease in time between initial and final tests
when compared to the acute and control groups.

Methods:
For this experiment, nine female and nine male mice of similar age will be selected from the
Loras College mouse room. Three mice of each gender will be randomly placed in the chronic,
acute, and control treatment groups. Each mouse will then be housed in an individual cage
marked with its treatment group and be fed as normal. First, the mice will be weighed to
determine the correct individual dosages of grape seed extract. Because 100 micrograms/g/day of
this extract in mice improves memory (Sarkaki, et. al., 2013), we will use this amount. To test
spatial memory, we will use the complex maze in Figure 1 because the mice will use proximal
cues to find the way to the end (Vorheese & Williams, 2014).
Figure 1. The mice will be placed in the spot marked with “x,” and the peanut butter will be at
the end with the arrow.

Second, each mouse will go through an initial trial of the maze before receiving treatment of the
extract. For all trials, the maze will be placed on the floor in a spot that can be seen from the
window in an adjacent room. Only a red light will be on to observe the mouse without disturbing
its dark cycle, and a one-half teaspoon of peanut butter will be placed at the bottom right corner
of the maze to motivate it to finish. Each mouse will be placed in the top left corner of the maze,
and the time it takes for them to complete it will be measured and recorded. Third, treatment of
the grape seed extract will begin for the next five days. Mice drink between four and seven
milliliters of water per day (“Mice – Food & Water,” n.d.); to ensure sufficient amounts are
provided, the individual dosages of extract will be dissolved in 12 milliliters of water per mouse
that will be changed every twenty-four hours. The control group will not receive any extract in
the water, and the chronic treatment group will have 100 micrograms/g/day. The acute treatment
group will receive regular water the first four days. On the fifth day, 500 micrograms/g of the
extract will be given to this group to receive the entire chronic treatment dosage at one time.
During these five days, the mice will be timed in one maze trial at the same time of day (Sarkaki,
et. al., 2013). On the sixth day, all mice will receive water without extract, and a final maze trial
will be conducted. A decrease in maze trial times over the week means the mice were able to
navigate the maze faster and ultimately improved their memories of it. If one group of the mice
has a larger difference in completion times than the others, then that means the treatment the
mice received enhanced their memory more.

List of Materials Needed:


● 18 mice
● One jar of peanut butter
● 250 mg of Grape Seed Extract
● Maze
● Water

References
Jamshidzadeh, A., Baigi, B. F. B., & Aram, M. (2010). The Effects of Grape Seed and Grape
Pomace Extracts on Spatial Memory Impairment Induced be Hyoscine in Mice. Journal
of Medicinal Plants Research, 4(22), 2334–2339.
Mice — Food & Water. (n.d.). Retrieved September 26, 2017, from
http://nswschoolanimals.com/mice/mice-food-water/
Praag, H. van, Lucero, M. J., Yeo, G. W., Stecker, K., Heivand, N., Zhao, C., … Gage, F. H.
(2007). Plant-Derived Flavanol (−)Epicatechin Enhances Angiogenesis and Retention of
Spatial Memory in Mice. Journal of Neuroscience, 27(22), 5869–5878.
https://doi.org/10.1523/JNEUROSCI.0914-07.2007
Sarkaki, A., Rafieirad, M., Hossini, S. E., Farbood, Y., Motamedi, F., Mansouri, S. M. T., &
Naghizadeh, B. (2013). Improvement in Memory and Brain Long-term Potentiation
Deficits Due to Permanent Hypoperfusion/Ischemia by Grape Seed Extract in Rats.
Iranian Journal of Basic Medical Sciences, 16(9), 1004–1010.
Subash, S., Essa, M. M., Al-Adawi, S., Memon, M. A., Manivasagam, T., & Akbar, M. (2014).
Neuroprotective effects of berry fruits on neurodegenerative diseases. Neural
Regeneration Research, 9(16), 1557–1566. https://doi.org/10.4103/1673-5374.139483
Vorhees, C. V., & Williams, M. T. (2014). Assessing Spatial Learning and Memory in Rodents.
ILAR Journal, 55(2), 310–332. https://doi.org/10.1093/ilar/ilu013
Wang, J., Ho, L., Zhao, W., Ono, K., Rosensweig, C., Chen, L., … Pasinetti, G. M. (2008).
Grape-Derived Polyphenolics Prevent Aβ Oligomerization and Attenuate Cognitive
Deterioration in a Mouse Model of Alzheimer’s Disease. Journal of Neuroscience,
28(25), 6388–6392. https://doi.org/10.1523/JNEUROSCI.0364-08.2008