Undergraduate Thesis Or Project
 

Immunohistochemistry Studies of Tumor Necrosis Factor Αlpha and Interferon Gamma Expression in Pseudocapillaria tomentosa Infected and Uninfected Zebrafish

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  • Tumor necrosis factor alpha (TNF α) and interferon gamma (IFN γ) were initially recognized as inflammatory mediators produced by immune cells in response to infectious agents or foreign material. (1,5) However, over the past decade much research in mammalian species has indicated that these proteins also play key roles in development and homeostasis of many tissues such as adipose tissue and endocrine islets of pancreas (2,3,12). Our goal was to determine whether the levels of these mediators were elevated in tissues of zebrafish following infection with intestinal nematodes. We examined protein expression for these mediators in whole histologic sections of zebrafish using immunohistochemistry applied to uninfected fish and fish infected with the intestinal nematode Pseudocapillaria tomentosa. Fish were sampled for histologic study at selected time points following infection with the nematodes. We utilized commercially available antibodies to mammalian TNF α and IFN γ and the DAKO Envision Plus Immunohistochemistry polymer system for detection of these proteins in fish tissues. Contrary to our initial hypothesis that parasite infected fish would have increased levels of these inflammatory mediators in tissues, we found no overall increase in these mediators in parasite infected fish using immunohistochemistry at the 12 week post-infection sample point. However, the early 5 week post-infection sample point showed that there was an increase in the levels of these mediators in parasite infected fish. Similar with observations in mammals that these mediators play important roles in tissue development and homeostasis, we found consistent, specific, moderate to strong baseline expression of these mediators in several tissues of zebrafish. In assays conducted at 5 weeks following parasite infection we saw increased levels of TNF α and IFN γ in inflamed skeletal muscle and saw increased TNF α in segments of bowel in which recent inflammation occurred.
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file details LoLaleeC2009.doc 2018-06-22 Pubblico Scaricare
file details Zebra_fish_TNF_A_and_IF_G.ppt 2018-06-22 Pubblico Scaricare