Journal of Undergraduate Biology Laboratory Investigations

2018

ETHANOL DECREASES HEART RATE OF

DAPHNIA COMPARED TO WATER
Gillian Cloar, Tram Dinh, Andrea Hoffen, Jordan Humphrey, Miranda Vesy*
1
University of Oklahoma, Department of Biology, 730 Van Vleet Oval, Room 314 Norman, OK 73019
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Various substances can have multiple effects on the number of action potentials taking place in a neuron, which
can directly impact the heart rate of a myogenic organism. Does ethanol alcohol decrease or increase action
potentials in a neuron? We tested this question using daphnia as our organism, making it unique because daphnia
are a myogenic organism, just like humans. Throughout five trials, we tested these organisms and 5% ethanol
combined with aquarium water, we found that ethanol alcohol caused the heart rate of the daphnia to decrease,
in this case, due to a decrease in action potentials in the organism.
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Introduction the same effect on all myogenic organisms, we

The leading cause of death in the United tested the effect of ethanol on Daphnia heart rate.
States is heart disease. As scientists have begun Although most arthropods have neurogenic hearts,
developing synthetic cardiac muscle cells, they have or hearts controlled by their nervous system,
run into a problem with getting the genetically daphnia are an exception to this because they are
engineered cells to beat at the same rate as the myogenic (Stein, 1966). This means that their hearts
original heart cells. By experimenting with different beat due to spontaneous heart muscle activity rather
substances, scientists hope to develop a drug which than in response to the pacemaker activity of the
can suppress or increase heart rate as needed (Shaw cardiac ganglion (Hill, 1992). Humans are also
& French, 2018). The heart beats because its’ myogenic, which makes daphnia an exceptional
pacemaker cells increase the membrane voltage candidate for testing the effects of drugs on the
enough to generate rhythmic action potentials. This heart (Stein, 1966.) They are also small and
then generates contraction of the heart muscle, relatively transparent, making the heart easily
commonly known as a heartbeat (DiFrancesco, accessible for viewing without using dissection or
2012). In a rat, ethanol was shown to decrease the other methods which would in turn harm the
number of heart beats per minute due to depressing organism. Using information from these past
action potentials in the membrane (Gimeno, 1962). journals, we have hypothesized that heart rate will
This decrease in action potentials is due to the decrease in Daphnia submerged in 5% ethanol
postsynaptic receptors being much less sensitive compared to Daphnia submerged in their original
(Wallner, 2003). Curious as to whether ethanol has aquarium water due to the fact that ethanol
depresses action potentials in the membrane. We
* Research Mentor
 JUBLI Volume: 1. Issue: 2 (2018)

will know that this hypothesis is supported if there Results

is a lower heart rate after daphnia submersion in An unpaired t-Test was conducted to
ethanol compared to water. However, a higher heart compare the effect of ethanol on daphnia heart rate.
rate after ethanol submersion would disprove our There was a significant difference between the two
hypothesis. groups; with the degree of freedom is 4, p= .0167.
Daphnia submerged in a solution of 5% ethanol
Methods showed a significant decrease in the average percent
To test the effect of ethanol on the heart rate change in daphnia heart rate compared to the
of daphnia, we used a solution of ethanol and daphnia in the control group, which were
aquarium water and a solution of 100% aquarium submerged only in aquarium water. However, even
water. We used the protocol for handling daphnia though ethanol had a significant effect on heart rate,
found on Canvas (Shaw & French, 2018). We there was not a consistent percent change between
recorded the daphnia under a microscope for 15 trials. There were outliers above and below the
seconds then watched this video in slow motion average percent change in the groups submerged in
while using a clicker to count the heart rate of the 5% ethanol, trial 3 (-8.76%) and trail 4 (-47.25%).
daphnia. By multiplying this number by four, we This means that although the percent change
calculated the heart rate in beats per minute, or between ethanol and aquarium water showed an
bpm. After this, we used a dropper to add one drop obvious difference, there is still significant
of 5% ethanol alcohol to the aquarium water difference between different daphnia in each trial
solution that the daphnia was submerged in on the because of independent heart rates of each daphnia.
slide. The daphnia was submerged in the When the daphnia were in 5% ethanol, the rate of
water/alcohol solution for seven minutes in order to their heart change percent decreased, on average, by
give the alcohol time to affect the daphnia. After the 21.70%, compared to when they were in water, as
seven minutes passed, the daphnia was again seen in Figure 1. When Daphnia was submerged in
observed under the microscope and heart rate was 5% ethanol, their heart rate decreased every time,
calculated in the same way that it was before the while the percent heart rate of Daphnia in water was
alcohol was added. Heart rate was recorded before overall consistent (Figure 1). Although in some
and after the ethanol was added in order to trials there was a slight decrease of heart rate and in
accurately measure the rate of change. We repeated other trials there was a slight increase, the change it
this using five daphnia in order to increase the heart rate had little to no change.
accuracy and size of our experimental group. As our
control, we repeated this entire process but with one
difference: rather than adding a drop of alcohol, a
drop of aquarium water was added. This sham
control was used to improve the accuracy of our
data by showing whether the alcohol affected the
heart rate or the heart rate was simply the time spent
in a slide, which obviously causes some stress on
the organism.
We now found the percentage rate of change
between the first and second observed heart rates in
each of the ten daphnia. These percentages were
then averaged and placed onto a bar graph for ease
of viewing. This was done using the software Figure 1. Average percent change decreasing
Microsoft Excel. Due to the fact that our data was rapidly in Daphnia’s heart rate in 5% ethanol
both nominal and measurement, and that we had compared to steady average percent change in
two unpaired groups, analysis is conducted by Daphnia’s heart rate in aquarium water.
running an unpaired T-test on the percent change
values.
 JUBLI Volume: 1. Issue: 2 (2018)

Discussion
As we hypothesized, the average heart rate of Gimeno, A. L., Gimeno, M. F., & Webb, J. L.
daphnia is significantly lower after 5% ethanol (1962). Effects of ethanol on cellular membrane
alcohol submergence, compared to submergence in potentials and contractility of isolated rat
aquarium water (as seen on figure 1). This atrium. American Journal of Physiology.
correlation was consistent throughout all five 203(1): 194-196.
daphnia in our experiment. Our control group, the
daphnia not submerged in ethanol, had no Hill, R. B., et al. "Nervous regulation of the
significant heart rate increase before and after the myogenic heart in early juveniles of the isopod c
seven minutes of sitting on the slide. The Fisher rustacean, Ligia exotica." Phylogenetic
study supports our conclusion that alcohol Models in Functional Coupling of the CNS
consumption decreases action potentials in and the Cardiovascular System: 3rd
myogenic organisms (Fisher, 1975). Because International Congress of Comparative
humans and daphnia are both myogenic, it is a Physiology and Biochemistry, Satellite
reasonable conclusion that alcohol decreases action Symposium, Shimoda, August 31-September
potentials, and therefore heart rate, in humans. 2, 1991. Vol. 11. Karger Medical and
As an alternative interpretation, one could Scientific Publishers, 1992.
argue that the decreased heart rate is more
dependent on the lab setting and. stress that the Kaas, B., Krishnarao, K., Marion, E., Stuckey, L., &
organism was placed under rather than the alcohol. Kohn, R. (2009). Effects of melatonin and
However, our control group goes to disprove this ethanol on the heart rate of Daphnia magna.
interpretation by showing no decrease in heart rate Impulse: the premier journal for
when no ethanol was used. Another alternative undergraduate publications in the
interpretation of our results could be that the vagus neurosciences.
nerve regulated the lowering of the heart rate, rather
than the alcohol consumption. Shaw, T. & French, D. (2018). Authentic Research
The reason for decreased action potentials in in Introductory Biology, 2018 Ed.
daphnia due to ethanol consumption is unknown, as Fountainhead, Fort Worth.
it could be decreased acetylcholine release, lowered
sensitivity of acetylcholine receptors, or a Stein, R. J., Richter, W. R., Zussman, R. A., &
combination of both (Kaas, 2009). As a future Brynjolfsson, G. (1966). Ultrastructural
experiment, the amount of acetylcholine released in characterization of Daphnia heart muscle.
the synapse could be measured in order to help The Journal of cell biology, 29(1), 168.
determine the exact reason for the decrease in action
potentials caused by ethanol alcohol. This could Wallner, M., H. J. Hanchar, and R. W. Olsen.
help to show how exactly what kind of heart "Ethanol enhances α4β3δ and α6β3δ γ-
problems could be combated with ethanol. aminobutyric acid type A receptors at low
concentrations known to affect humans."
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