MINNELLAA19
HOSRIA19
Cell Communication Lab
Purpose-
The purpose of this lab was to explore how cell communication using two strains of
yeast.
Background-
Both unicellular and multicellular organisms use cell communication in order to elicit a
response which helps an organism coordinate and respond to their environment. Cellular
communication can occur through direct contact, local signaling, or long-distance signaling. For
a response to occur, a message much reach a receptor that is able to receive that type of message
which then turns on the ability for many other reactions, or a cascade. Pheromones are a type of
signaling molecule that is secreted from one organism and detected from another organism thus
providing various messages through this cellular communication and eliciting a change.
In this lab we focused on the mating of yeast. Yeast is a unicellular eukaryote that
undergoes many of the same biological processes as humans. In the yeast cell cycle, cell growth
and cell division are tightly linked and are dependent on factors such as nutrient concentration.
Depending on environmental cues, yeast can undergo asexual or sexual reproduction to produce
new cells. The type of yeast that mate are haploids, which contain one copy of the genome, like
egg or sperm cells. There are two haploid mating types, Mat a and Mat alpha, and these cells can
bud and reproduce asexually, like diploid yeast. The mixed mating types cells can become a
haploid, budding haploid, zygote, budding zygote, or a shmoo Each of these mating types release
pheromones. During this phase, two haploids continue to grow towards each other until
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achieving cell-cell contact. The zygote formed then re-enters the mitotic cell cycle, which gives
rise to the first diploid bud. In this lab we are going to try and view yeast reproduction through
cell communication.
Procedure-
Petri dishes and culture tubes were labeled “a-type” and “alpha-type”. Then, 3 mL of
broth were added to each culture tube and the specific strains of yeast was placed into the
corresponding culture tubes. Transferring the yeast colonies into the tubes was accomplished by
using a toothpick to scrape yeast colonies from the subculture dishes. The yeast suspensions were
then placed, using a pipette, onto their respective plates and spread over the surface of the agar.
The agar plates were then incubated for 24 hours. Wet mounts were then created for the “a-type”,
“alpha-type”, and a mixed culture. Each slide was observed under different fields of
magnification at various time intervals. However, the time intervals were unfortunately not
optimal due to a shortage of time.
Data-
HAO Type Culture Single Haploid Budding haploid
Time 0 Field 1 (40) 22 16
field 2 (40) 18 9
field 3 (40) 25 23
total per stage 65 48
percent of total 58% 42%
time 1 field 1 (40) 74 19
field 2 (400) 72 31
field 3 (40) 80 27
total per stage 226 77
percent of total 74.60% 25.40%
time 2 field 1 (400) 79 52
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field 2 (40) 77 47
field 3 (400) 82 61
total per stage 238 160
percent of total 59.80% 40.20%
time 3 field 1(400) 100 62
field 2 (40) 96 53
field 3 (400) 102 66
total per stage 298 181
percent of total 62.20% 37.80%
Budding
HBO type culture Single Haploid haploid
Time 0 Field 1 (40) 3 0
a few minutes apart from eachother field 2 (40) 1 2
field 3 (40) 1 1
total per stage 5 3
percent of total 62.50% 37.60%
time 1 field 1 (40) 15 20
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after 15 min field 2 (400) 25 40
field 3 (40) 16 22
total per stage 56 82
percent of total 40.60% 59.40%
time 2 field 1 (400) 800 130
after 2hr field 2 (40) 600 115
field 3 (400) 815 131
total per stage 2216 376
percent of total 85.50% 14.50%
time 3 field 1(400) 805 135
after 2hr and 15 min field 2 (40) 610 210
field 3 (400) 821 134
total per stage 2236 389
percent of total 85.10% 14.80%
Mixed Single Budding Single Budding
Culture Haploid Haploid Shmoos Zygotes Zygotes Asci
Field 1
Time 0 (100) 1256 96 4 188 20 20
Field 2 212 8 12 16 8 0
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(400)
Field 3 (40)
total per
stage 1468 104 16 204 28 20
percent of
total 79.80% 5.70% 0.87% 11.10% 1.50% 1.10%
Field 1
Time 1 (100) 1184 84 8 72 18 16
Field 2
(400) 84 24 12 16 12 4
Field 3 (40)
total per
stage 1268 108 20 88 30 20
percent of
total 82.70% 7.00% 1.30% 5.70% 2.00% 1.30%
Field 1
Time 2 (100) 744 78 8 56 32 12
Field 2
(400) 44 4 0 12 4 0
Field 3 (40)
total per
stage 788 82 8 68 36 12
percent of
total 79.30% 8.20% 0.80% 6.84% 3.60% 1.20%
Field 1
Time 3 (100) 560 60 4 32 28 12
Field 2
(400) 80 8 4 4 0 0
Field 3 (40)
total per
stage 640 68 8 36 28 12
percent of
total 80.10% 8.60% 1.00% 4.50% 3.50% 1.50%
- no data could be
collected/view of
magnification was not
optimal
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Conclusion-
Through this lab, we were able to observe cells communicating. Our data also proves that
we were able to see cell communication as it is modeled. The mixed culture reproduced sexually
and the isolated culture reproduced asexually. We know that the isolated cultures reproduced
asexually because only single haploids and budding haploids were present. At each time interval
for each culture there were more single haploids than budding. The HAO strains single haploid
to budding haploid percentage was 75% to 25%, and the percentage for the HBO strain was 85%
to 15%. The cells were reproducing at a very slow rate asexually. The mixed culture was a lot
different since sexual reproduction occurred and we
could view the different stages of life and view the haploids becoming gametes, shmoos were
also produced. A shmoo looks like a pear, and it the two cells combining together. Seeing the
haploids become gametes proved to us that the yeast colony was reproducing sexually. There was
difficulty in counting the very dense yeast colony but the differences in the development of cells
reproduction were clear. Shmoos production proved sexual reproduction because before a shmoo
is formed a signal cascade must occur. In the A type and Alpha type secreted pheromones which
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attracted them to each other and allowed for them to mix and reproduce. The shmoo would not
have been created without cell communication or pheromones because the pheromones initiated
the signal cascade after binding to a receptor of the opposite strain. Without the pheromones
binding to the cells receptor no communication would have occurred to signal reproduction and
create a shmoo to form in the yeast colony.
Although this experiment was successful and helped us observe cell communication
through our microscopes there were some sources of error that may have occurred. One of the
major issues was trying to collect accurate data. It was extremely difficult to count each of the
full fields from the microscopes so we split the fields up into quadrants and counted what was in
that view then multiplied by four. This method seemed like the most efficient since counting a
full field would become very tedious and it is even easier to lose your place counting all of the
tiny bubbles. Counting the fields of view was a huge difficulty in collecting our data but being
able to distinguish the various stages of the cells reproduction through the picture we took of the
microscope. Our last problem was time, we not only did not have enough time to collect our
isolated cultures and had to work after class time to complete them but the lab data also had to be
collected at specific time intervals which were a bit hard to keep consistent while trying to
collect data throughout. Although there were many sources of error that could have affected our
data we still came out with a result that helped us observe and understand cell communication
better.
Appendix-
1. No, there was no visible difference between the two although they are genetically
different.
2. Yes, the mixed cultures clearly had cells at different life stages and shmoos were clearly
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visible. The isolated cultures, however, did not have gametes like the mixed cultures, so
various life stages and shmoos were not present.
3. This lab proved that cells can communicate via pheromones. Communication was
observed without direct contact through the pheromone signaling in the mixed cultures.
The shmoos were formed and were able to fuse together as a result of the opposite cells
sending pheromones which bound to the receptors on the opposite cell.
4. 4. Colonies C and D would develop shmoos first because the pheromones secreted from a
yeast cell are attracted to the opposite strain of yeast. The pheromones from colonies C
and D will reach the opposite strain first, causing the colonies to develop into shmoos.
5. The purpose of the 24-hour incubation period was to allow the yeast cells to develop into
colonies. No, you would not expect to observe the same changes in cellular morphology
if the cultures were not incubated.
Big Idea
1. The unknown colony could be added to a known colony of type-a yeast. Then you would
observe the life stages of the yeast cells after a period of time to determine if shmoos
developed. If shmoos did not develop, then the unknown is type-a. However if shmoos
developed, then the unknown is type-alpha because shmoos only develop when
pheromones bind to the receptors of the opposite yeast culture.
2. A pheromone reaches the opposite type yeast cell and binds to its receptors, and a signal
cascade is triggered. The signal cascade can then regulate the transcription of genes
which allow for the growth of the cell towards the direction of the opposite cell which
forms a shmoo.
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