Investigating Cell Migration Under Microgravity Conditions Co-Principal Investigators: Sylvia Edoigiawerie, Kamsi Odinammadu, Amelia Smith, Beverly Wu Teacher Facilitator: Dr. Michelle Starz-Gaiano University of Maryland, Baltimore County University System of Maryland, Maryland 2016 SSEP National Conference Smithsonian Air and Space Museum, Washington, DC June 29-30, 2016

Cell Migration is Essential (if you are in the animal kingdom) Human Embryo http://www.primobaby.com Development www.ehd.org Immune Cell Migration Wound Healing and Immune Response http://en.wikipedia.org/wiki/File:Laceration,_leg.jpg http://makeagif.com/I0Z2VD

Overview: The Process of Cell Movement Actin (cytoskeletal protein) → facilitates movement E-cadherin (membrane protein) → involved in cell to cell adhesion Cells migrate by responding to chemical signals in their environment that trigger them to polymerize monomers of a protein called actin and to extend the cell like an arm at a side of the of the cell known as the leading edge. At the leading edge, the cell then adheres to the surface it moves on. Contraction at the cell body and de-adherence at the trailing end, which pushes the rest of the cell forward. This same migratory process occurs in border cells in the oocyte of fruit flies, our model system. Pictured above are stage 8-10 fo the migratory process

Integrative Model of Cell Migration Actin (cytoskeleton) Activating protein E-Cadherin (adhesion) We incorporate an integrative perspective to analyze cell migration Cell locomotion

Cells in the Fly Ovary are a Model for Migration Advantages: fantastic genetic tools small transparent tissues border cells In adult female flies, the ovary occupies the majority of the abdominal cavity. The ovary pair is made up of about 30 ovarioles, shown here, which are strings of developing egg chambers at successively older stages of development as you move from anterior to posterior. Each of these units will develop into a single egg. The egg chamber is made up of 16 large cells in the center, which are the germ line cells that will give rise to the embryo, and about 600-900 somatic follicle cells, which surround the germ line cells in an epithelial layer and give rise to the egg shell. Today we will focus on a small subset of the follicle cells, the border cells, indicated in blue, which arise within the epithelial layer and then begin a dramatic migration. Let’s look more closely these 3 stages of development, which are most relevant to border cell migration…. blue= nurse cell nuclei ovariole (L. Manning)

Does gravity affect the number of cells that become motile or their ability to complete movements? Activator

Experimental Plan http://swampthingscience.com/?author=1 www.hansoltc.com www.e-reagent.com https://www.thinglink.com Flies engineered to make migratory cells fluoresce green 1 sex per FME section with fly food – open to allow mating Use chemical fixative to freeze cells in place Observe and score ovaries

Analysis and Interpretation microgravity predictions Disorganized F-Actin? F-Actin Lathienna Manning, Starz-Gaiano Disrupted or decreased E-cadherin? (normally actin binding stabilizes E-cadherin) Due to previous literature that shows alterations in actin remodeling networks under microgravity conditions in immune cells, we speculate that actin dynamics in our analogous system will also become disorganized under microgravity conditions. Due to actin’s close relationship with another protein E-cadherin, which is required for cell-cell adhesion, we also expect E-cadherin phenotype to deviate from wildtype. microgravity predictions E-cadherin Niewiadomska, P., D. Godt, and U. Tepass.

Future Directions Grounds for more research on the impact of microgravity on immunity A focus on astronaut health in space conditions grows in importance as space exploration expands

Acknowledgements NCESSE Starz-Gaiano Laboratory University of Maryland, College Park University of Maryland, Baltimore County Maryland Space Grant Consortium Department of Cell Biology and Molecular Genetics, UMD CP The Drosophila Community for Fly Stocks Bloomington Stock Center Funding for the Starz-Gaiano lab from National Science Foundation