University Honors

To understand how lung inflammation alters parts of the brain that regulate breathing, weuse Alternaria alternata, an air-borne fungal allergen, and observe how it affects the NucleusTractus Solitarii (NTS) region of the brain. The NTS is the major regulatory region for breathing.To imitate the air-borne quality and the effects of A. alternata, we expose mice to ultrafineaerosolized non-infectious particles of A. alternata in controlled chambers for one week.Exposure is done in conditions where the mice live in their home cage without any restrictions intheir moving, sleeping, or eating patterns. This administration mimics the most common route ofexposure in mammals. After exposure, NTS brain samples are collected and cryosectioned 5micrometers, then used to quantify molecules involved in the neuronal synapse. We incubate theNTS brain sections with immunofluorescent antibodies specific to VGLUT2, an excitatorypresynaptic marker, and PSD-95, a postsynaptic marker. We use immunofluorescence confocalmicroscopy to produce 3D imagery of the molecules. We quantified Vglut2/Syn double positivepuncta and observed a sex specific decrease in presynaptic sites. We sought to quantify thepostsynaptic using PSD95, however the sections we used did not pass quality control because thelevel of staining and the image capture was oversaturated. Secondly, we sought to see if thedecrease in excitatory presynaptic sites was due to a decrease in the gene expression or adifference in localization. Using NanoString methodology, we could see that there were nosignificant changes in the entire brainstem. At the level of the total brainstem RNA, there was nochange in the expression of PSD95, VGLUT2 or EAAT2. We cannot preclude that there is adecreased expression locally. Future studies will be quantified by digital spatial profiling. Thisproject will help determine the effect that allergic lung inflammation has on the brain regulatingbreathing.