Leah Wheaton
Pronouns: She/her
Research Mentor(s): Benjamin Singer
Research Mentor School/College/Department: Internal Medicine / Pulmonary and Critical Care Medicine / Medicine
Program:
Authors: Leah Wheaton, Kate Giffin, Grace Roughton, Michael Newstead, Benjamin Singer
Session: Session 3: 11:00 am – 11: 50 am
Poster: 91
Abstract
Sepsis is associated with mental illnesses and cognitive deficits. Synapse loss is a mechanism of cognitive deficits in neuropsychiatric disorders and the hippocampus is a key brain area for cognition. Our goal is to see if the number of hippocampal synapses decreases in mice that have undergone sepsis compared to uninfected mice. A secondary goal of the project is to improve immunofluorescent staining of synapses by comparing paraformaldehyde and glyoxal fixation. The brains from mice that either underwent sepsis or were uninfected were fixed by paraformaldehyde or glyoxal and sliced to 40 micrometers on a cryostat. We used immunofluorescence to stain brain tissue for the synaptic markers Homer1 and Bassoon in the hippocampus. Homer1 is a postsynaptic protein and Bassoon is a presynaptic protein. Then the tissue was imaged on the Stellaris confocal microscope and synapses, defined as puncta formed by the overlapping of Homer1 and Bassoon signals, will be quantified in each group using FIJI image analysis software. We found that glyoxal fixation improves staining for synaptic markers compared to paraformaldehyde fixation. We have successfully stained one set of tissue from female mice 18 hours after abdominal infection, which is when they are in the acute illness stage, and analyses are underway to quantify the synapses present and whether the synapse numbers differ between sepsis and control groups. We plan to repeat these experiments in male mice and at later time points after infection. This research could help aid in the progression of treatments to combat the neuropsychological effects of sepsis after recovery.
