Location
D.P. Culp Center Ballroom
Start Date
4-5-2024 9:00 AM
End Date
4-5-2024 11:30 AM
Poster Number
128
Name of Project's Faculty Sponsor
Allan Forsman
Faculty Sponsor's Department
Health Sciences
Competition Type
Competitive
Type
Poster Presentation
Presentation Category
Health
Abstract or Artist's Statement
All tissues used in this study were generated in a study in the laboratory of Dr. Johnathan Peterson. The study was designed to determine the relationship between C1q TNF-related protein-3 (CTRP3) and non-alcoholic fatty liver disease. It is estimated that over 3 million Americans suffer from non-alcoholic fatty liver disease. This disease also has a higher prevalence in persons with Type II diabetes and various cardiac pathologies. Transgenic mice bred to over express CTRP3, were generated by mating female C57 mice with male, 4513+ mice. Littermates without the CTRP3 transgene were used as wild-type (WT) controls. All mice used in the study were fed a very high fat diet (VHFD). Mice were 6 weeks old at the onset of the study and were maintained on the VHFD for 12 weeks. At the end of the 12-week feeding period the mice were sacrificed, their organs harvested and placed in 10% formalin for overnight fixation. Following fixation, the tissues were placed in 70% ETOH for storage until paraffin embedding. Tissues were dehydrated and embedded using standard embedding techniques. The embedded tissues were then sectioned at 4 µm and mounted on microscope slides. The slides were then stained using a standard H&E staining method. Matching slides were stained using Masson’s Trichrome staining. The slides were viewed under standard light microscopy to evaluate the histology/pathology of the tissue. Representative tissue samples from each treatment group were also subjected to immunohistochemical staining to confirm the presence of proteins indicative of membranes associated with lipid vacuoles. Since daily injections of CTRP3 have been shown to reduce high-fat diet-induced fatty liver, we hypothesize that overexpression of CTRP3 will have a protective effect on fat deposition in the livers of VHFD mice. To date, our data have not indicated a strong positive correlation between overexpression of CTRP3 and reduction of fat deposition in the liver of VHFD mice. WT mice fed a VHFD exhibited expected levels of fat deposition in their livers. Of the mice overexpressing CTRP3, approximately half of them appeared to have reduced fat levels compared to their wild type controls, while approximately half of them exhibited high levels of fat deposition in the liver. Other studies found CTRP3 to be protective against fat deposition in the livers of VHFD mice. Those studies differ from ours, such that, in those studies, CTRP3 overexpression was accompanied by daily injections of CTRP3.This could be an explanation for the unexpected results. Further, since our mice were generated by mating female C57 mice with male, 4513+ mice, the litters were either WT or CTRP3 transgenic, as the transgene only needs one copy, we will look at Western blot data to confirm that each animal in the study (WT vs VHFD) was correctly genotyped. At the current time, our data suggests that we reject our hypothesis and conclude that over expression of CTRP3 does not protect the liver from fat deposition and fat related pathologies in mice fed a VHFD.
Transgenic overexpression of CTRP3: Possible Protection against Non-Alcoholic Fatty Liver Disease
D.P. Culp Center Ballroom
All tissues used in this study were generated in a study in the laboratory of Dr. Johnathan Peterson. The study was designed to determine the relationship between C1q TNF-related protein-3 (CTRP3) and non-alcoholic fatty liver disease. It is estimated that over 3 million Americans suffer from non-alcoholic fatty liver disease. This disease also has a higher prevalence in persons with Type II diabetes and various cardiac pathologies. Transgenic mice bred to over express CTRP3, were generated by mating female C57 mice with male, 4513+ mice. Littermates without the CTRP3 transgene were used as wild-type (WT) controls. All mice used in the study were fed a very high fat diet (VHFD). Mice were 6 weeks old at the onset of the study and were maintained on the VHFD for 12 weeks. At the end of the 12-week feeding period the mice were sacrificed, their organs harvested and placed in 10% formalin for overnight fixation. Following fixation, the tissues were placed in 70% ETOH for storage until paraffin embedding. Tissues were dehydrated and embedded using standard embedding techniques. The embedded tissues were then sectioned at 4 µm and mounted on microscope slides. The slides were then stained using a standard H&E staining method. Matching slides were stained using Masson’s Trichrome staining. The slides were viewed under standard light microscopy to evaluate the histology/pathology of the tissue. Representative tissue samples from each treatment group were also subjected to immunohistochemical staining to confirm the presence of proteins indicative of membranes associated with lipid vacuoles. Since daily injections of CTRP3 have been shown to reduce high-fat diet-induced fatty liver, we hypothesize that overexpression of CTRP3 will have a protective effect on fat deposition in the livers of VHFD mice. To date, our data have not indicated a strong positive correlation between overexpression of CTRP3 and reduction of fat deposition in the liver of VHFD mice. WT mice fed a VHFD exhibited expected levels of fat deposition in their livers. Of the mice overexpressing CTRP3, approximately half of them appeared to have reduced fat levels compared to their wild type controls, while approximately half of them exhibited high levels of fat deposition in the liver. Other studies found CTRP3 to be protective against fat deposition in the livers of VHFD mice. Those studies differ from ours, such that, in those studies, CTRP3 overexpression was accompanied by daily injections of CTRP3.This could be an explanation for the unexpected results. Further, since our mice were generated by mating female C57 mice with male, 4513+ mice, the litters were either WT or CTRP3 transgenic, as the transgene only needs one copy, we will look at Western blot data to confirm that each animal in the study (WT vs VHFD) was correctly genotyped. At the current time, our data suggests that we reject our hypothesis and conclude that over expression of CTRP3 does not protect the liver from fat deposition and fat related pathologies in mice fed a VHFD.