Independent Research
for Bio 371/372 academic credit during the fall or spring semester
or summer research with paid stipend
Dr. Karen Hales, Dana 201,
Phone x2324, email kahales
Dr. Hales's home page
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Summer 2013 schedule

Research focus: In the Hales lab we perform genetic analysis with the fruit fly Drosophila melanogaster to explore the molecular mechanisms by which mitochondria are moved and shaped in cells. Mitochondria are the organelles often referred to as the "powerhouses of the cell," since they are the sites where energy from food is stored in ATP. In many cell types with unusual energy needs, mitochondria move (in a regulated way) to be close to energy-requiring structures such as flagella or ion pumps. In addition, mitochondria often undergo regulated fusion and division, sometimes existing as a single large network in the cell, and sometimes as many individual units. We speculate that such changes may contribute to the efficiency of ATP generation in certain contexts. In Drosophila melanogaster (as in most other higher organisms), mitochondria undergo a dramatic series of shape changes during spermatogenesis. Through the identification of male-sterile mutants defective in mitochondrial morphogenesis, we can isolate and characterize genes whose protein products function in this process.

Current personnel in the lab:
Bethany Wagner '14
Eric Sawyer '14
Devon Harris '15
Nishita Suresh '14
Hannah McMillan '15
Lindsay Regruto (Research technician)
James Winkle (Research technician)
Tucker Bates '15 (returning Spring 2014)

Past projects:
Fitz Sturgill '01 Inverse PCR to identify a candidate gene for no mitochondrial derivative
Shaheen Counts '01 Characterizing Drosophila male sterile mutants with mitochondrial defects in spermatids
Sean Burke '02 Identification of a candidate gene for no mitochondrial derivative (nmd) in Drosophila melanogaster.
Marisa Wilson '02 Characterization of new mitochondrial mutants in Drosophila
Laura Quillian '03 Investigation of mitochondrial morphogenesis in Drosophila: recombination mapping of two new mutants with defects in nebenkern formation.
Monica Siegenthaler '03 Generation of germline clones to define a role for the milton gene in mitochondrial morphogenesis in Drosophila spermatids.
Sarah Baxley '04 Testing the nmd candidate gene for rescue of the mutant phenotype
Sara Holmberg '04 Characterization of the subcellular localization of the nmd gene product
Ty Plowshay '05 Characterization and fine mapping of the mitoshell mutant
Lauren Watson '06 EMS screen for new nmd alleles
Lucy Marcil '06 Characterization of new mitochondrial mutants in Drosophila
Laura Sedig '07 Characterization of the mitoshell phenotype and generation of deficiencies in the mitoshell region
Ben Whigham '07 Determining the role of milton in Drosophila spermatogenesis
Carrie Black '06 Characterization of the specificity of a polyclonal antibody raised against Nmd
Kevin Saunders '05 Characterization of the specificity of a polyclonal antibody raised against Nmd
Lindsay Nakaishi '05 Generating new nmd alleles
Anna Nam '08 Characterizing the subcellular localization of the nmd gene product
Levi Benson (Colgate '06, summer REU student) Generating germline clones to dissect the roles of milton and dMiro in Drosophila spermatogenesis
Sarah Durnbaugh '06 Characterizing the subcellular localization of the nmd gene product (Honors project)
Kara Koehrn '06 Generation of mutants in the CG4701 gene in Drosophila
Sarah Coffey '09 Determination of microtubule morphology in mitoshell and nmd
Bevin English '08
Characterization of the hypomorphic male sterile nmdry4 allele and analysis of the nmd paralog CG4701’s putative role in mitochondrial morphogenesis during Drosophila spermatogenesis.
Katie Wood '08 Cloning and characterization of mitoshell, a gene required for normal mitochondrial aggregation during Drosophila spermatogenesis
Laura Bergner '09 The role of beta tubulin and other proteins in mitoshell-mediated cytokinesis events in Drosophila spermatogenesis
Mike Beaucaire '09 The effect of Sib mutations on cytokinesis failure in Drosophila spermatogenesis

Carolyn Wakeman '09 Characterizing nuclear morphology and cytokinetic defects in mitoshell, a recessive male sterile line ofDrosophila melanogaster
Kin Lau '10 Determining if the mutation which causes sterility in the AA2-37 and AA2-73 strains of Drosophila melanogaster is on CG4701
Eddie Hickman '09 Characterizing mitoshell: localizing the gene product, sequencing new alleles, and determining
effects on centriole and basal body placement
Laura Bergner '09 Dissecting a novel mutant: characterizing temporal and cytokinetic defects in the mitoshell mutant
Hunter Stone '09 Testing whether CG4701rescues male sterile mutations in the AA273 and AA237 male sterile lines of Drosophila
Madeleine Pilon '12 Construction of a mitoshell rescue construct
Sarah Pyfrom '11 Dissecting gene disregulation in mitoshell
Dylan Coughtrey '11 Subcellular localization of the CG4701 gene product
Lauren Ivey '11 Testing candidate genes for ms(2)1400
Tessa Campbell '11 Assessing premature mitochondrial fusion in mitoshell
Casie Genetti '11 Assessing functionality of Nmd-GFP fusion protein
Leah Smith '13 Assessing basal bodies in CG4701 male sterile mutants
Taylor Gunnell '11 Tracking mitochondria in ms(2)1400
Sarah Pyfrom '11 Characterizing the function of nmd in Drosophila spermatogenesis
Dylan Coughtrey '11 Characterizing the function of CG4701 in Drosophila spermatogenesis
Lauren Ivey '11Constructing a ms(2)1400-GFP fusion transgene
Casie Genetti '11 Roles of ATP synthase subunits in mitochondrial shaping in Drosophila spermatogenesis
Tessa Campbell '11 Assessing the ms(2)1400 phenotype in the absence of mitochondrial fusion
Chris Lima '11
Characterizing the emmenthal mutant phenotype in Drosophila spermatogenesis
Conroy Field, Johnson C. Smith University '14, RT-PCR to assess transcriptional defects of CG7813 in ms(2)1400 mutants
Julie Coursen '14 Visualizing microtubules in nmd mutants
Aaron Deal '12 Characterizing the emmenthal mutant phenotype in Drosophila spermatogenesis
Yihharn Hwang '12 Confirming that CG7813 corresponds to ms(2)1400
Melissa Lorenzo '12 Testing roles for ATP synthase subunits in Drosophila spermatogenesis
Laura McCoy '13 Testing expression levels of a candidate gene for ms(2)1400 by RT-PCR

William Mitchell '13 Characterizing the molecular defect in emmenthal male sterile mutants
Olivia Brown '13 Testing roles for ATP synthase subunits in Drosophila spermatogenesis
Jessica Gerard '14 Characterizing structural defects in CG4701 male sterile mutants of Drosophila
Shaina McCaskill, Fayetteville State University '12 Characterizing contractile ring defects in nmd and CG4701 male sterile Drosophila
Conroy Field, Johnson C. Smith University '14) Testing roles for ATP synthase subunits in Drosophila spermatogenesis
Kelsey Sheaffer '13
Testing roles for ATP synthase subunits in Drosophila spermatogenesis
Megan Bannon '14 Computational analysis of ATP synthase subunits enriched in the Drosophila testis

Oshauna Morgan, JCSU '16 Role of the testis-enriched g subunit of ATP synthase in mitochondrial shaping during Drosophila spermatogenesis

 

Information and requirements for students enrolled in Bio371 or 372

Course objectives

In this independent research course you will learn to design and carry out experiments to explore the molecular basis of mitochondrial movement and shaping in developing fruit fly sperm cells. You will learn basic techniques of fly husbandry, microscopy, and/or recombinant DNA technology. You will become familiar with reading the research literature. You will develop skills as an experimentalist, proposing new experiments to follow up on recent results, and learning to troubleshoot techniques.

Textbooks/Reading
There are no official textbooks. We will discuss journal articles during our weekly meetings. You will participate in literature searches and paper selection in preparation for these journal club-style discussions.

Expectations
--A two page project proposal at the beginning of the semester
--Attendance at weekly lab meetings
--At least ten hours a week of research time in the lab. The ten hours do not include time in lab meeting or time filling out your time sheet.
--Formal presentation of your results (with powerpoint) every three or four weeks during lab meeting
--Informal discussion of your results with the group every week during lab meeting
--Weekly submission of a progress sheet indicating daily activities, achievements, troubleshooting, and goals.
--Frequent one-on-one informal conversations with me as needed
--Literature searches and background paper reading as applicable to your project
--Poster creation and subsequent presentation at the Spring Science and Math Research Symposium.
--A write-up detailing your methods and results.

Grading

Proposal

Data presentations

Journal club

Poster

Overall effort and output (calculated as average of weekly interim grades)

5%

15%

15%

15%

50%

 

WWW resources
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Literature browsing/searching

Browse current journals
Search Pubmed
Genetics news

Drosophila resources

A Drosophila
Fly husbandry guidelines by Grace Panganiban
Fly protocols from WWW Virtual Library
FlyBase at Indiana University
Berkeley Drosophila Genome Project
Bloomington Drosophila Stock Center
Interactive Fly
FlyAtlas
WWW Virtual Library: Drosophila
Drosophila labs on the web
Annual Drosophila Research Conference
Drosophila news group

Mitochondria resources

Mitochondrial Research Society
Mitochondrial Medicine Society
United Mitochondrial Disease Foundation
Children's Mitochondrial Disease Network

 

Sequence analysis

NCBI Main Page
BLAST search for similar nt or aa sequences
CLUSTALW Multiple Sequence Alignment
Transmembrane domain prediction
PSORT prediction of subcellular localization

 


Labs working on Drosophila spermatogenesis

Chris Bazinet (St. Johns)
Julie Brill (Hospital for Sick Children, Toronto)
Steve DiNardo (University of Pennsylvania)
Margaret Fuller (Stanford; follow link on dept page)
Maurizio Gatti (Universita' di Roma "La Sapienza")
Erika Matunis (Johns Hopkins)
Angus McQuibban (University of Toronto)
Steve Wasserman (UC San Diego)
Helen White-Cooper (Cardiff University)

Labs working on mitochondrial morphogenesis in yeast

Rob Jensen (Johns Hopkins)
Jodi Nunnari (UC Davis)
Liza Pon (Columbia)
Janet Shaw (University of Utah)
Peter Thorsness(University of Wyoming)
Benedikt Westermann (Universität Bayreuth)

Labs working on mitochondrial morphogenesis in other systems

Pascale Belenguer (Université Paul Sabatier)
David Chan (CalTech)
David Logan (Université D'Angers)
Mark McNiven (Mayo Clinic)
Alex van der Bliek (UCLA)

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© Copyright 2002-2013 Department of Biology, Davidson College, Davidson NC 28035
last modified August 25, 2013 by K. Hales