NATIONAL CENTER FOR CASE STUDY TEACHING IN SCIENCE

Speciation
and the
Threespine Stickleback
by
Joan C. Sharp1, Erin M. Barley1,
Kevin K-W. Lam1, Suraaj Aulakh1,
Allison Cornell2, and Kathleen A. Fitzpatrick1

Introduction
Watch the following video on allopatric speciation: <https://youtu.be/TVv7om3WE_M> (running time: 13:32 min).
This video describes the process of speciation that occurs when a population is split in two by a geographical barrier.
After separation, each population accumulates genetic differences through the four mechanisms of genetic divergence
(founder effect, genetic drift, mutation, natural selection). If the two populations reunite, there are three possible
outcomes: (1) they are a single species, (2) they are separate species, or (3) they are in the process of speciating.
In this case study, we examine a fascinating fish called the threespine stickleback. We focus on Paxton Lake, a lake
where two populations of these fish that were once geographically separated came together again. In Part I, we ask
What is the outcome of this reintroduction?

Part I – Background
Threespine sticklebacks (Gasterosteus aculeatus) are minnow-sized fish that live in coastal marine environments
throughout Europe, Asia, and North America. Marine threespine sticklebacks are named for the three dorsal spines
that help protect them against predators. In addition, they have bony lateral plates to protect their sides and pelvic
spines to provide protection on their underside. The dorsal and pelvic spines are held erect by a bony structure called
a pelvic girdle. The sides of threespine sticklebacks are well protected with an armor of bony lateral plates. Marine
sticklebacks filter-feed on zooplankton, using the long bony gill rakers that line their throats and sieve out small prey
from the water that flows into their mouths and over their gills.
In coastal areas, many freshwater lakes and streams are also home to populations of sticklebacks that have become
adapted to local freshwater conditions. One such lake is Paxton Lake on Texada Island, British Columbia. Texada is
one of the Gulf Islands in the Salish Sea between the British Columbia mainland and Vancouver Island. The origin
of the freshwater sticklebacks on Texada Island dates back to glaciation around 13,000 years ago, when the weight of
glaciers lowered coastal land relative to sea level and the current Gulf Islands were submerged. As glaciers melted and
retreated, the land rebounded and the islands emerged from the ocean, trapping water in lakes that gradually became
fresh over time. Throughout the Gulf Islands, freshwater lakes are home to threespine stickleback populations that
became trapped and isolated from marine populations as the Gulf Islands formed.

1
Department of Biological Sciences, Simon Fraser University, Burnaby, BC.
2
Department of Biological Sciences, Cedar Crest College, Allentown, PA.

Case copyright held by the National Center for Case Study Teaching in Science, University at Buffalo, State University of New York. Original-
ly published January 3, 2019. Please see our usage guidelines, which outline our policy concerning permissible reproduction of this work. Image
credit: A selection of sticklebacks,by Alexander Francis Lydon (1836–1917), pd, <https://archive.org/details/britishfreshwate01houg/page/n7>.
 NATIONAL CENTER FOR CASE STUDY TEACHING IN SCIENCE

But the story gets more interesting; Paxton Lake is home to not one but two different forms of threespine sticklebacks.
The evidence supports a double invasion hypothesis. This proposes that after the first marine population of sticklebacks
was trapped and became adapted to local conditions in the lake, additional marine sticklebacks made their way up-
stream from the ocean into the lake. Paxton Lake is now home to two related but distinct populations of sticklebacks,
both descended from a marine ancestor.
In Paxton Lake, the two populations of sticklebacks—limnetic and benthics—are named for the areas of the lake in
which they live and feed. Limnetic sticklebacks feed on zooplankton in the limnetic zone, the open waters of the lake
near the surface and away from shore. Benthic sticklebacks feed on larger invertebrates in the benthic zone, the muddy
lake bottom in the shallow water near the edge of the lake.
Limnetic sticklebacks are slim with narrow mouths. Like their marine ancestors, they have a protective armor of dorsal
and pelvic spines, pelvic girdle, and many lateral plates. This armor offers protection from predatory cutthroat trout,
which feed on sticklebacks in the limnetic zone of the lake. Their throats contain large numbers of long gill rakers,
used by limnetic sticklebacks to filter tiny zooplankton out of the water.
Benthic sticklebacks are stocky fish, with deep bodies and wide mouths. In contrast to their marine ancestors, the ben-
thic fish aren’t as well protected. Most have only two dorsal spines instead of the usual three. Most lack pelvic spines
and a pelvic girdle and have few lateral plates. They have a relatively small number of short gill rakers and do not filter
feed. Instead, they feed on bottom dwelling organisms such as worms, clams, and insect larvae in the muddy shallows
of the benthic zone. Their main predators in the benthic zone are large insects such as backswimmers and dragonfly
larvae, which capture young sticklebacks by seizing their spines.
Figure 1 shows a limnetic male and a benthic
male. Limnetic and benthic sticklebacks live, feed,
and face predation in different areas of Paxton
Lake. However, males of both types build nests,
court females, and care for their eggs side-by-side
in the benthic zone of the lake. Sticklebacks have
long been studied for their interesting mating
behaviors. In springtime, male sticklebacks build
nests of vegetation glued together by sticky secre-
tions. A mating male develops bright colors and
becomes very aggressive, swimming out to defend
his nest from all attackers. A researcher studying Figure 1. Limnetic male stickleback (top) and benthic male stickleback
the sticklebacks described how the little males (bottom). Image Credit: Elizabeth Carefoot, Learning and Instructional
would swim up and try to nip at her as she waded Development Centre, Simon Fraser University. Used with permission.
into their territories!
When a female stickleback enters a male’s territory, he recognizes her by her swollen belly, full of eggs. Instead of
attacking, the male courts the female, performing a zigzag dance or nipping at her fins. If the female is receptive, she
follows him to his nest. After she enters the nest, the male nudges her to encourage her to spawn. He then enters the
nest himself and releases sperm to fertilize the eggs. The male guards the fertilized eggs until they hatch, oxygenating
them by fanning the nest with his pectoral fins.
The threespine sticklebacks of Paxton Lake have been studied by researchers at the University of British Columbia
(UBC) for over 25 years, first by Dr. Don McPhail and more recently by a research team headed by evolutionary
biologist Dr. Dolph Schluter. In this case study, you will design experiments that are similar to those carried out by the
UBC researchers and will interpret the data generated from their experiments.
The following video introduces you to Dr. Schluter, the researcher who led much of the research that you will be
examining: <https://youtu.be/FRwqspBKX-I> (running time: 4:52 min).

“Speciation and the Threespine Stickleback” by Sharp, Barley, Lam, Aulakh, Cornell, & Fitzpatrick Page 2
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Part II – Interaction Between Benthic and Limnetic Sticklebacks

In Part I, you learned about the background and history of Paxton Lake’s stickleback fish and specifically about the
two distinct populations of sticklebacks that are now in contact. We now ask which of the three outcomes is occurring:
(1) benthic and limnetic sticklebacks are one species; (2) benthic and limnetic sticklebacks are separate species; (3)
benthic and limnetic sticklebacks are in the process of speciating. Below are data from UBC researchers to help us
answer this question.

Survival of Benthic, Limnetic, and Hybrid Sticklebacks

Method: Artificially fertilized eggs from benthics, limnetics, and hybrids were reared in the laboratory.
Results: Table 1. Survival of fertilized eggs to hatching.
Form Total # of Eggs Survivors / Egg # % Survival
Benthics 682 662/682 97%
F1 hybrids 659 626/659 95%
Limnetics 599 575/599 96%
F2 hybrids 525 504/525 96%

Source: McPhail, J.D. 1992. Ecology and evolution of sympatric sticklebacks (Gasterosteus): evidence for a
species pair in Paxton Lake, Texada Island, British Columbia. Canadian Journal of Zoology 70: 361–9.

Questions
1. What can we conclude from the data in Table 1?
FZ hybrids have the least amount of eggs and survivors yet
they have a similarsurvival rate with Limnetics
2. What is the difference between F1 and F2 crosses, and why are both crosses necessary?
FI may be a male Benthic breeding with a female Limnetic,
while FZ may be a mate Limnetic breeding with a female Benthic
Both important as the PNA shared may vary based on males and
Proportions of Benthic, Limnetic, and Hybrid Stickleback in Paxton Lake females in the species'
Method: Large numbers of fish were captured from Paxton Lake during the summers of 1969, 1979, and 1986. Table
2 shows the proportions of benthic, limnetic, and hybrid fish captured from the lake each year.
Results: Table 2. Proportions (and numbers) of benthic, limnetic, and hybrid sticklebacks in summer trap samples.
Year Total Benthics Limnetics Hybrids
1 1057 0.50 (528) 0.48 (509) 0.019 (20)
2 962 0.50 (479) 0.49 (473) 0.010 (10)
3 994 0.49 (491) 0.49 (489) 0.014 (14)

Source: McPhail, J.D. 1992. Ecology and evolution of sympatric sticklebacks (Gasterosteus): evidence for a
species pair in Paxton Lake, Texada Island, British Columbia. Canadian Journal of Zoology 70: 361–9.

Question
3. What can we conclude from the data in Table 2?
this shows that there may be low inter-breeding in
the wild for hybrids and thus a significantly lower
population compared to Benthics and Limnetics despite having
“Speciation and the Threespine Stickleback” by Sharp, Barley, Lam, Aulakh, Cornell, & Fitzpatrick
high fertility Page 3
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Morphology of Benthic, Limnetic, and Hybrid Sticklebacks

Table 3. Comparison of anatomical differences between laboratory-reared benthic, limnetic, and F2 hybrid sticklebacks.
Form Benthic Limnetic F2 Hybrids
Average body depth: 10.6 mm 8.8 mm 9.7 mm
Percentage of population with 56% 2% 46%
two (vs. three) dorsal spines:
Percentage of population 81% 0.2% 37%
without pelvic girdle:
Average number of gill rakers: 18.7 23.8 20.6
Average gill raker length: 0.8 mm 1.5 mm 1.2 mm
Average jaw width: 3.7 mm 2.5 mm 3.2 mm
Average number of lateral plates: 0.7 11 6.9

Source: McPhail, J.D. 1992. Ecology and evolution of sympatric sticklebacks (Gasterosteus): evidence for a species pair
in Paxton Lake, Texada Island, British Columbia. Canadian Journal of Zoology 70: 361–9.

Questions
4. What can we conclude from the data in Table 3?
FZ hybrids share roughly half of the anatomical
difference percentages between n Benthic and Limnetic sticklebacks

5. Given the data from Tables 1–3, which of the three possible outcomes from renewed contact between isolated
populations best describes the interaction between benthic and limnetic sticklebacks? In other words, do they form
one species, separate species, or are they in the process of speciating?

From the three possible outcomes from renewed

contact between isolated populations, they
are best described as in the process of speciating

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