THE EVOLUTION OF STICKLEBACK POPULATIONS Randi Fielding, Section 028 Lab # 9

Section One: Introduction

According to ARKive, a small fish able to live in both saltwater and freshwater that has sharp spines along its back in front of the dorsal fin is called a threespine stickleback. Threespine sticklebacks can be organized into three distinct population categories: marine, sea-run and freshwater (Leady, 2014). Each of the populations is different based on the environment the sticklebacks can be found in. Sticklebacks in the marine form live and breed strictly in the ocean while freshwater stickleback live entirely in fresh water (Leady, 2014). Sea-run sticklebacks spend most of their lives in the ocean, however, they are anadromous so freshwater is where they migrate to breed (Leady, 2014). Scientists have discovered, by measuring allele frequencies, that some of the stickleback populations present in lakes today differ from those of the past, as they have mutations which may have helped them evolve quickly between saltwater and freshwater forms (Nature, 2012). Sea-run sticklebacks, as a result of being trapped in postglacial lakes, had to evolve in order to continue living in their new freshwater environment (Leady, 2014). This means the sea-run stickleback populations have gradually been forced to change and develop over generations to adapt to their new surroundings by reducing or eliminating their pelvic girdles and spines for a lighter form better suited for freshwater (Leady, 2014). As the sticklebacks remained trapped in the postglacial lakes, this led to the changes in the distribution and frequencies of alleles or, in other words, evolution (Science Daily, 2013). According to Science Daily, stickleback fish are good for studying evolution and changes in population because they are a small fish with short generational time periodsmaking themclearly readable and easily visible. Since they are so small, they are a lot of fossilized sticklebacks available and easy to locate making living sticklebacks readily comparable with fossilized sticklebacks. In this particular lab experiment, sticklebacks from two lakes around Cook Inlet, Alaska (Bear Paw Lake and Frog Lake) and fossil sticklebacks from the Truckee Formation in Nevada, a site where 10 million years ago a lake existed, were evaluated to answer the question Do changes in body structure reflect evolution? The hypothesis is that the pelvic girdle and pelvic spines will change over generations. This means that the skeletal structure will either remain unchanged thereby having a complete pelvic girdle and spine, or their skeletal structure will either be reduced or absent, both of which indicate there have been an evolutionary change in their pelvic structures (Leady, 2014). My prediction is that specific changes will be found in the two (2) experiments that indicate evolution is occurring.

Section Two: Methods

The experiment performed is listed in Lab #9, found in the Fundamentals of Life Science Lab Manual (Leady, 2014). There were no changes made to the protocol listed.

Section Three: Results

The results for Experiments #1 and #2 are shown in both of the tables and graphs below. The analysis of the pelvic girdles and spines show that evolution is occurring because changes in the stickleback skeletal structure were found. By studying the stickleback fish, either living or fossilized, sticklebacks can be divided into three distinct categories based on their pelvic skeletal structure: (1) Complete sticklebacks with a complete pelvis and pelvic spine, (2) Reduced sticklebacks with the pelvis reduced to one or two smaller bones and no pelvic spine or (3) Absent no pelvis or pelvis spine (Leady, 2014). The third table demonstrates the results of other data provided by other researchers in terms of additional fossilized layers that were studied.

Experiment #1: Living Stickleback Fish in Bear Paw Lake and Frog Lake
# OF THE STICKELBACK 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 TOTAL COMPLETE TOTAL REDUCED TOTAL ABSENT BEAR PAW LAKE Reduced Reduced Absent Reduced Absent Reduced Reduced Reduced Reduced Reduced Reduced Reduced Reduced Absent Reduced Reduced Reduced Reduced Absent Reduced 0 Complete 16 Reduced 4 Absent FROG LAKE Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete 20 Complete 0 Reduced 0 Absent

Experiment #2: Fossil Stickleback Fish from the Truckee Formation in Nevada
# OF THE STICKLEBACK 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 TOTAL COMPLETE TOTAL REDUCED TOTAL ABSENT LAYER 2 Complete Complete Complete Complete Reduced Reduced Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete Complete 18 Complete 2 Reduced 0 Absent LAYER 3 Absent Reduced Reduced Reduced Absent Reduced Reduced Reduced Reduced Reduced Reduced Reduced Reduced Reduced Absent Absent Reduced Reduced Reduced Absent 0 Complete 15 Reduced 5 Absent

Results of Fossil Fish Pelvic Structure Analysis LAYER 1 LAYER 2 LAYER 3 LAYER 4 LAYER 5 LAYER 6 20 18 3 1 0 0 COMPLETE 0 2 16 19 15 16 REDUCED 0 0 1 0 5 4 ABSENT
* This research includes data provided by other researchers for the additional layers not used within this experiment.

Section Four: Discussion

Do changes in body structure reflect evolution? The hypothesis is that the pelvic girdle and pelvic spines of the stickleback fish will change over generations. My prediction is that specific changes will be found in the two (2) experiments that indicate evolution is occurring. The results of the experiment, listed above, show that evolution has occurred as the stickleback fish analyzed, both living and fossilized, indicate a reduction or absenteeism in pelvic girdles and pelvic spines which is indicative that an evolutionary change took place in their pelvic skeletal structure in response to remaining in the freshwater environment. In Experiment #1, living stickleback fish from two lakes around Cook Inlet, Alaska, specifically Bear Paw Lake and Frog Lake, were collected to determine whether there were significant differences between the two populations (Leady, 2014). 20 sticklebacks were taken from each lake, preserved and then stained with a solution that turns bones within them red so that the skeletal structures can be analyzed (Leady, 2014). The pelvic structures were then scored complete, reduced or absent and compared against one another and to different marine phenotype in order to determine whether or not possible environmental factors influenced the changes in their skeletal structure. The two lakes vary because Bear Paw Lake prevents predatory fish from readily entering the lake since it is completely enclosed and does not drain at the surface (Alaska Department of Game and Fish, 2013). Frog Lake, on the other hand, allows access by predatory fish to the lake because it is connected by a stream to the water system (Alaska Department of Game and Fish, 2013). In examining the data collected, the results support the idea that different predators in the two lakes influence pelvic structures. In Bear Paw Lake, where predatory fish are prevented from readily entering, the sample of stickleback fish examined showed that out of 20 stickleback fish, 16 were scored reduced and 4 were scored absent. This demonstrates that an evolutionary change has taken place in Bear Paw Lake because stickleback fish have a reduced skeletal structure or absent a skeletal structure (no pelvic girdle or spine). Frog Lake, on the other hand, out of 20 stickleback fish that were analyzed, 20 were scored complete. Sticklebacks present in Frog Lack and have not changed their skeletal structures because their pelvic girdles and spines are still necessary for protection against the predatory fish that can inhabit Frog Lake via the stream to the water system. In Experiment #2, fossilized stickleback fish that lived 10 million years ago from the Truckee Formation in Nevada, a lake that existed about 10 million years ago, were analyzed to determine the change in stickleback fish over time (Leady, 2014). The layers analyzed in the experiment were layers 2 and 3 and data was provided by other researchers for additional layers. All of the data was spanned over the course of 15,000 years in order to make an estimation in pelvic structures for this population (Leady, 2014). 20 sticklebacks from Layer 2 were analyzed, scoring 18 complete and 2 reduced. 20 sticklebacks from Layer 3 were analyzed, scoring 15 reduced and 5 absent. The further down in layers,

the further back in time the layers go making Layer 2 older than Layer 3 and proving that the sticklebacks have evolved. As the number of complete pelvises changed over the years (Layer 1 20, Layer 2 18, Layer 4 1, Layer 5 0, Layer 6 0), this shows there was a relative decrease in the frequency of fossilized stickleback fish with complete pelvic girdles and spines. According to the data provided by other researchers for additional layers, the pelvic structure changed very quickly at within the first half of the years analyzed, down to 3 complete stickleback fish being verified. The data shows that towards the end of the 15,000 years, roughly in the last 7,500 years, that only one complete stickleback was verified and moving towards present day 0 complete stickleback fish were verified. Looking at the fossilized stickleback fish can help to calculate the rate of change over time, helping in analyzing the evolutionary change in the population. The change over the years was able to be studied and proven that evolution has occurred because the information needed was preserved in the fossilized layers. The results from Experiment 2s table listed above support the same idea.

Section Five: Summary

In conclusion, the results showed that specific changes in the two experiments were found to indicate that evolution is occurring, as most of the stickleback shrimp scored had changes to their pelvic skeletal structure as a response of adapting and changing to their freshwater environment. The only weakness in Experiment #1 could be by the experimenter wrongly scoring the stickleback shrimp. It would provide incorrect data and an incorrect analysis if the pelvic skeletal structures was wrongly identified. In Experiment #2, the sticklebacks were fossilized and that could provide a weakness if the fossils were damaged, deteriorated or decayed. If the fossils incurred any of the aforementioned, the pelvic structures being scored could have deteriorated or decayed where they might be scored as reduced or absent when, in fact, they were complete before such occurred. During this lab, since it was a virtual lab completed on the computer, there were no problems in completing the experiment.

Citations
ARKive. (2003-2013). Three-spined stickleback (Gasterosteus aculeatus aculeatus). Retrieved 20 October 2013 from http://www.arkive.org/three-spined-stickleback/gasterosteus-aculeatus-aculeatus/ Leady, B. (2014). Fundamentals of Life Science Lab Manual. Plymouth, MI: Hayden-McNeil Publishing. Nature International Weekly Journal of Science (2013.) Stickleback genomes reveal path of evolution. Retrieved 19 October 2013 from http://www.nature.com/news/stickleback-genomes-reveal-path-ofevolution-1.10392 ScienceDaily (2013). Stickleback Genome Holds Clues to Adaptive Evolution. Retrieved 21 October 2013 from http://www.sciencedaily.com/releases/2012/04/120404133753.htm State of Alaska (2013). Alaska Department of Fish and Game Northern Cook Inlet Management Area. Retrieved 21 October 2013 from http://www.adfg.alaska.gov/index.cfm?adfg=ByAreaSouthcentralNorth CookInlet.main