CASE TEACHING NOTES for “Those Old Kentucky Blues: An Interrupted Case Study” by Celeste A. Leander, Departments of Botany and Zoology, University of British Columbia Robert J. Huskey, Department of Biology, University of Virginia INTRODUCTION / BACKGROUND The purpose of this case, based on an extended family from the Appalchian Mountains which had a high occurrence of the rare disorder methemoglobinemia, is to provide students with an example of a Mendelian inheritance pattern that can be viewed as either dominant/recessive or as incompletely dominant, depending on perspective. The case has been used in first year biology in both traditional and honors sections. Typically, first-year science majors have misconceptions about inheritance. The case presents students with a seemingly clearcut example of a dominant/recessive inheritance pattern. However, when examined from the phenotype of enzyme function, it becomes obvious that this is not clear cut and in fact looks to be an example of incomplete dominance. Both are correct. It simply depends on the perspective from which the phenotype is being analyzed. In the first case, the phenotype is skin color. In the second case, the phenotype is enzyme activity level. Objectives After completing this case, students will understand: • Hypothesis generation. • Pedigree construction and analysis. • Evaluation of inheritance patterns. • Reconciliation of changing inheritance patterns, depending on perspective. CLASSROOM MANAGEMENT This case is designed for small groups of four to six students. Each group is responsible for generating answers to all of the questions. I pass out the first part of the case and allow the students five minutes to read it and answer the first set of questions. I then open up the floor for five minutes for group discussion. I keep tally on the board. Student hypotheses based on Ruth’s observations typically fall under two broad headings: environmental causes (including contaminated food or water) or genetic causes. It is helpful to mention that methemoglobinemia can be congenital or acquired by exposure to various chemicals (such as some antibiotics and anesthetics), so both categories of hypotheses are reasonable. Armed with this information, I then ask the students to report on a way to test these hypotheses. Some will want to do invasive blood tests, test the water in the area, etc. I then reveal what Ruth did by introducing Part II of the case. I mention to the students that Ruth knew that if blueness is a heritable trait, it would pass through a lineage in a predictable pattern. I pass out the second part of the case and allow the students minutes to read it and generate a pedigree. Each group draws their pedigree on the board. Once the pedigree is drawn, students are asked to add genotypes for all individuals. Any conflicts should be pointed out and discussed among the groups. I walk around and pose questions to the students at this stage. If students are on the right track, I C T N for “Those Old Kentucky Blues” by Leander & Huskey Page may ask, “How come Elizabeth is heterozygous?” Sometimes groups will fixate on an inheritance pattern that doesn’t work. An example is sex-linked inheritance. In this case, I may ask, “How come Luna is blue?” Many students will assign John’s genotype as homozygous dominant (“BB”) because none of his children were blue. Other students will argue that we can’t know that for sure. This is a good chance to point out that each child is an independent statistical event, so we cannot in fact know that John is homozygous dominant. This discussion and group work typically lasts to minutes. Once everyone is satisfied with the pedigree, students are sent back to their seats comfortable with the decision of a dominant/recessive inheritance pattern for “blueness.” I then pass out the NADH diaphorase activity graph. After a few minutes, I ask the students to articulate what this graph says. They will typically agree that the parents (heterozygotes) are somewhere in between the two homozygous phenotypes. This is an example of incomplete dominance. The difference is that we are no longer analyzing “blueness” as a phenotype; our perspective has changed. We are now analyzing enzyme activity as our phenotype. I give them minutes to answer the questions from Part II in their groups. Students are usually uncomfortable with the idea that an inheritance pattern is not set in stone but can change depending on the phenotype analyzed. I leave the last five minutes of class open for questions and clarification. To wrap up the discussion, I mention that a specific, yet unrelated, mutation in a hemoglobin protein can also cause congenital blue coloration of the skin. This mutant form of hemoglobin does not allow NADH diaphorase to interact properly with the iron atom. In this case, the mutation is inherited as a dominant phenotype with regards to skin color. A heterozygous person appears blue because hemoglobin is a structural protein; when half of the hemoglobin is non-functional, it becomes evident phenotypically. Enzymes, however, are effective in very small concentrations. Thus, the mutation in NADH diaphorase, as presented in this case, is recessive when evaluating a heterozygote because having just half of a regular dose of the enzyme is enough to get the job done. BLOCKS OF ANALYSIS Detailed case analysis is provided in a separate file that is password-protected. To access this information, go to the detailed case analysis. You will be prompted for a username and password. If you have not yet registered with us, you can see whether you are eligible for an account by reviewing our password policy and then apply online or write to answerkey@sciencecases.org. Contested Geneology It should be noted that the family history part of the Science article (Trost, ) on the blue people of Troublesome Creek does not match well with the evidence compiled by professional genealogists. There was a Martin Fugate who married Elizabeth Smith (October , ), but there is no evidence that any of their children were “blue.” At least one record indicates that they had children, not seven. One of their children is listed in one source as Zachariah Fugate, but he married Margaret Triplett, not a “Smith” sister of Elizabeth. One of the children of Martin and Elizabeth was the Levi Fugate (born about and died about ) who married a “Ritchie,” namely Mahala “Cricket” Ritchie (born June , , and died July , ). This couple had seven children including Luna Fugate (born June and died in ). There was another Martin Fugate whose wife was named Mary, but her maiden name is not known. This Martin Fugate, a son of Benjamin Fugate and Hannah Devers, was born about in Russell County, Virginia, and died about in Breathitt County, Kentucky. Martin and Mary Fugate had seven children including Zachariah “Ball Creek Zach” Fugate and Martin “Big Man” Fugate. C T N for “Those Old Kentucky Blues” by Leander & Huskey Page Zachariah “Ball Creek Zach” Fugate married Mary “Polly” Smith who was a sister to the Elizabeth Smith who married the first Martin Fugate mentioned. “Ball Creek Zach” and “Polly” had children including two who were known to be “blue”—John “John Blue” Fugate and Lorenzo Dow “Blue Anze” Fugate. It would seem that in the reconstruction of the family line through family stories, hazy memories, and guesses there was some confusion about which Zachariah belonged to which Martin. We may never be able to construct a totally accurate pedigree of this interesting family, but it is important to clear up what confusion we can. REFERENCES Fugate, Mary. The Fugate Family Newsletter Nov. . Vol. XVII, pg. . (ISSN: -) Trost, Cathy. The Blue People of Troublesome Creek. Science . November . http://www.brown.edu/Students/MAT//zowers/downloads/Standard_Artifact.pdf and http://www.rootsweb.com/~kyperry/Blue_Fugates_Troublesome_Creek.html Last accessed: January , . Umbreit, Jay. Methemoglobin—It’s Not Just Blue: A Concise Review. American Journal of Hematology ():–. . Acknowledgements: This case study was published with support from the National Science Foundation under Award . Any opinions, findings and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. Copyright © by the National Center for Case Study Teaching in Science. Originally published March , at http://www.sciencecases.org/blue_people/blue_people_notes.asp. Please see our usage guidelines, which outline our policy concerning permissible reproduction of this work. C T N for “Those Old Kentucky Blues” by Leander & Huskey Page