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Inflammation in Obesity and Molecular Engineering of a Transgenic Mouse Model of Diabetes

Al-Dieri, Ali Ghalib
http://rave.ohiolink.edu/etdc/view?acc_num=mco148131707089233

Abstract Details

2016, Doctor of Philosophy (PhD), University of Toledo, Medicinal Chemistry.
In the first project of type 2 diabetes and obesity, we investigated the role of macrophages, adipocytes, and cytokines in diet-induced metabolic syndrome. Multiphoton microscopy of adipose tissue from obese mice detected macrophages with lipid in crown-like structures surrounding adipocytes. Macrophages expressed both TNFa and IL-10. Adipocytes occasionally expressed IL-10, but not TNFa. Adipose tissue from obese mice was digested and the adipocyte floating layer was purified. The floating layer had high mRNA levels of TNFa, but surprisingly also contained a high level of F4/80 mRNA indicating the presence of macrophages. Investigation by confocal microscopy revealed that macrophages remain adherent to adipocytes in the floating layer and contain lipids in their cytoplasm. The majority of the adherent macrophages expressed TNFa, while the adipocytes did not. Therefore, our data do not support previous reports showing TNFa expression in adipocytes. We have showed that contamination by macrophages in the purified adipocyte floating layer explains the TNFa expression. Type 1 diabetes mellitus (T1D) is an autoimmune disease that is characterized due to the destruction of the beta cells of the pancreas, by the body’s immune system. Both CD4 and CD8 T cells are required for optimal disease transfer to occur, therefore, T1D, in NOD mice, is believed to be a T cell mediated autoimmune disease due exclusively to T cells that can transfer diabetes. It has been shown that by expressing a high density of insulin receptor (IR) in their surface, chinese hamster ovary (CHO) cells could migrate toward an insulin gradient. Thus the IR is a chemotactic molecule capable of cell movement toward an insulin gradient. Our aim was to determine if a high density of IR expression is a viable mechanism to deliver T cells to the islet regardless of antigen specificity and in strains of mice that do not normally become diabetic. Therefore, in the second project of type 1 diabetes, we steered towards a Cre-Lox system with a strong CAG viral promoter and an eGFP reporter that will drive a high IR expression on all T cells. This should illustrate expression of large amounts of exogenous, tagged IR while T cells are labeled with eGFP. To this end, a viable plasmid was constructed by subcloning a mIR fragment into a lox vector with CRE recombinase restriction to ensure specific expression.
Marcia McInerney, Ph.D (Advisor)
Herman von Grafenstein, Ph.D (Committee Member)
Zahoor Shah, Ph.D (Committee Member)
Michael Morran, Ph.D (Committee Member)
Imam Shahnawaz , Ph.D (Committee Member)
146 p.
Biochemistry; Biology; Immunology; Molecular Biology; Molecular Chemistry; Pharmacy Sciences

Recommended Citations

Citations

  • Al-Dieri, A. G. (2016). Inflammation in Obesity and Molecular Engineering of a Transgenic Mouse Model of Diabetes [Doctoral dissertation, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=mco148131707089233

    APA Style (7th edition)

  • Al-Dieri, Ali. Inflammation in Obesity and Molecular Engineering of a Transgenic Mouse Model of Diabetes. 2016. University of Toledo, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=mco148131707089233.

    MLA Style (8th edition)

  • Al-Dieri, Ali. "Inflammation in Obesity and Molecular Engineering of a Transgenic Mouse Model of Diabetes." Doctoral dissertation, University of Toledo, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=mco148131707089233

    Chicago Manual of Style (17th edition)

mco148131707089233
512
© 2016, all rights reserved.
This open access ETD is published by University of Toledo Health Science Campus and OhioLINK.