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INQUIRY & An Authentic Research Experience INVESTIGATION for Undergraduates in the Developmental Biology and Physiology Laboratory Using the Chick Embryonic Heart • JACQUELINE S. McLAUGHLIN, MIT A. PATEL ABSTRACT 2014; Lopatto, 2003; Seago, 1992). A four-step pedagogical frame- The lab presented in this paper utilizes a proven four-step pedagogical work that includes all essential elements of authentic research has framework (McLaughlin & Coyle, 2016) to redesign a classic Association of been developed that can serve to simplify and streamline the develop- Biology Laboratory Education (ABLE) undergraduate lab (McLaughlin & ment and implementation process in an introductory biology labora- McCain, 1999) into an authentic research experience on vertebrate four- tory setting (McLaughlin & Coyle, 2016). This framework has been chambered heart development and physiology. The model system is the shown to improve student perceptions of science and research, skills chicken embryo. Through their research, students are also exposed to the and knowledge levels in scientific research practices set forth by the embryonic anatomy and physiology of the vertebrate heart, the electrical National Research Council (2000), and confidence to effectively circuitry of the developing heart, and the effects of pharmacological drugs on heart rate and contractility. Classical embryological micro-techniques, engage in scientific research practices including the ability to write explantation of the embryo, surgical removal of the beating heart, isolation strong scientific lab reports and read scientific papers (McLaughlin of the heart chambers, and more advanced tissue culture methods are also et al., 2017). These results are consistent with other studies which conducted. In this laboratory paradigm, students work in pairs to ask their measured students’ perceptions and attitudes (Brownell et al., 2012; own questions concerning the effects of two human cardiovascular drugs, Harrison et al., 2011; Caruso et al., 2009; Weaver et al., 2008; ™ ™ denopamine and acebutolol on both in vivo and in vitro chicken Howard & Miskowski, 2005), ability to design experiments and inter- embryonic heart rate and contractility, develop testable hypotheses based on pret data (Myers & Burgess, 2003), and ability to develop information information gathered from relevant scientific literature, devise and carry out fluency (Gehring & Eastman, 2008) in laboratory courses that inte- a controlled experiment, and present the data in a professional scientific manner pertaining to a topic of clinical significance. grated authentic research-based experiences into the curriculum. Course-based The chicken embryo is a classic model Key Words: authentic research; heart development; organism used to illustrate the principles of chicken embryo; developmental biology laboratory; undergraduate physiology laboratory; undergraduate research. basic vertebrate embryology primarily because research experiences it is easily accessible, inexpensive, and direct observation of a living embryo is possible even Introduction (CUREs) provide a at the early stages of embryogenesis. With the aid of a dissecting microscope, the key stages Course-based undergraduate research experi- way to increase and of heart development and the circulatory pat- ences (CUREs) provide a way to increase broaden terns can be observed in vivo, even when the and broaden participation of students in chicken embryo is still attached to the yolk. authentic research (Auchincloss et al., 2014; participation of Another advantage to using the live chicken Bangera & Brownell, 2014), and train them embryo to demonstrate the principles of heart in the essential elements of authentic students in authentic development is that the embryo can be research, which include reading scientific lit- research. removed from the shell and maintained in vitro erature, generating questions (that don’t cur- in a temperature controlled environment for rently have answers), forming hypotheses, designing experiments, several hours while the beating of the heart, flow of blood, and collecting and analyzing data, working toward significant findings, the developmental stages accompanying the formation of the four and presenting results in both oral and written forms (Spell et al., chambers are even more clearly visible. In addition, the chicken The American Biology Teacher, Vol. 79, No 8, pages. 645–654, ISSN 0002-7685, electronic ISSN 1938-4211. © 2017 National Association of Biology Teachers. All rights reserved. Please direct all requests for permission to photocopy or reproduce article content through the University of California Press’s Reprints and Permissions web page, www.ucpress.edu/journals.php?p=reprints. DOI: https://doi.org/10.1525/abt.2017.79.8.645. THE AMERICAN BIOLOGY TEACHER CHICKEN EMBRYONIC HEART 645 embryonic heart is large enough for properly trained students to surgically isolate it from the embryo. Thus, students who work with this particular embryo in their development and/or physiology lab- oratories not only learn principles of vertebrate mor- phogenesis, organogenesis, and cardiac physiology, but are also exposed to simple, yet useful micro- manipulative techniques. These particular dexterous manipulations allow many of those who aspire to become physicians to try their hands (no pun intended) at surgical technique. We thoroughly redesigned a classic cookbook Figure 1. Illustration of human heart organogenesis from (A) a “tubular Association of Biology Laboratory Education (ABLE) heart” to (B) an S-shaped tube with the prominent ventricle bulging to the undergraduate lab on vertebrate heart development right (Gray, 1918/2000). © 2000 copyright Bartleby.com, Inc. (McLaughlin and McCain, 1999) to incorporate the above highlighted four-step laboratory pedagogical framework. This allows students to carry-out a higher level inquiry-based, authentic research experience (McLaughlin & Coyle, 2016; Goedhart & McLaugh- lin, 2016; McLaughlin et al., 2017) using present-day cardiovascular used as a model system for human heart development. The hearts drugs and updated tissue culture techniques and equipment. Stu- of the human and chicken embryos develop from the fusion of dents also master more advanced developmental biology, cardio- paired precardiac mesodermal tubes (aka “endocardial tubes”) vascular physiology, and pharmacology concepts. located on either side of the developing foregut, on the ventral sur- face. These paired tubes begin to fuse at the anterior end (head) and continue to fuse posteriorly to form one continuous tube Lab Protocol known as the tubular heart (Figure 1a). Once formed, the tubular heart is completely ventral to the foregut, and it has five distinct Developmental and Physiological Aspects of the regions that can be identified from anterior to posterior: truncus Chicken Embryonic Heart: Chronotropic and arteriosus, bulbus cordis, primitive ventricle, primitive atrium, and Inotropic Effects of Select Drugs sinus venosus (Figure 1b). Blood flows anteriorly, from the sinus venosus to the truncus arteriosus (future ascending aorta and pul- monary trunk). The heart begins to beat just after the paired heart Objectives tubes begin to fuse. It is the sinus venosus that becomes the future embryonic pacemaker. The key objectives in this lab are three-fold: (1) to think critically The tubular heart then elongates on the right side, looping and about, and understand, how the four-chambered vertebrate heart bending to form an S shape, with the prominent ventricle bulging develops using the chicken embryo as a model system; (2) to think to the right (Figures 1b and 2). The heart continues to fold upon critically about specific pharmacological agent(s) and their modes itself, moving the sinus venosus and atrium to a position anterior of action(s) at both the cellular and organ (heart) levels; and (3) to carry out a scientific research experiment. To accomplish these objectives, you will have eight weeks to carry out a four-step pro- cess wherein you will design and execute an authentic research project. This experimental framework is consistent with the man- ner in which professional research scientists design, execute, inter- pret, and communicate their experimental results. The over-arching experimental question that you will be attempting to answer through your research is the following: How do the cardiovascular drugs, denopamine™ and acebutolol™, affect the in vitro heart rate (chronotropic effect) and force of contraction (inotropic effect) of the developing isolated chicken heart? Background Knowledge The development of the four-chambered vertebrate heart involves a series of cellular migrations, fusions, and specific differentiations— that is, a multitude of morphogenetic events. Although gestation of Figure 2. Head and S-shaped heart of a developing chick the human and chick differ, their four-chambered hearts develop in embryo at 1.5 days (~36 hours), viewed from the ventral surface a very similar fashion, and because of this the chicken embryo is (Gray, 1918/2000). © 2000 copyright Bartleby.com, Inc. 646 THE AMERICAN BIOLOGY TEACHER VOLUME. 79, NO. 8, OCTOBER 2017 Figure 3. The developmental stages of a human heart beginning with the fusion of two endocardial tubes up to specific compartmentalization. Illustration of development of heart by OpenStax is licensed under CC by 3.0. and dorsal to the ventricle and the bulbus cordis (Figure 3). The ven- Step 2. Design an Experiment tricle is now U-shaped and in the medial