1.Effects of Dietary Fatty Acid Composition on Pro -and Macro -Glycogen Utilization and Resynthesis in Rat Skeletal Muscle.
Jongsam LEE ; Jaechul KIM ; Youngwoo KWON ; Jangkyu LEE ; Jeongpil LEE ; Chungsoo YOON
The Korean Journal of Nutrition 2007;40(3):211-220
The purpose of this study was to investigate that the effect of dietary fatty acid composition on pro- and macro-glycogen utilization and resynthesis. The analyses were further extended for different muscle fibers (type I, type II, & type IIb ) as well as tissues (i.e., liver & heart ). Total one hundred sixty Sprague-Dawley rats were used, and rats were randomly allocated into four experimental groups :animals fed standard chow diet (n =40 ), animals fed saturated fatty acid diet (n =40 ), animals fed monounsaturated fatty acid (n =40 ), and animals fed polyunsaturated fatty acid (n =40 ). Animals in each groups were further divided into five subgroups :sacrificed at REST (n =8 ), sacrificed at immediately after 3 hr swim exercise (P-0HR, n =8 ), sacrificed at one hour after 3 hr swim exercise (P-1HR, n =8 ), sacrificed at four hour after 3 hr swim exercise (P-4HR, n =8 ), and sacrificed at twenty-four hour after 3 hr swim exercise (P-24HR, n =8 ). Soleus (type I ), red gastrocnemius (type IIa ), white gastrocnemius (type IIb ), liver, and heart were dissected out at appropriated time point from all animals, and were used for analyses of pro- & macro-glycogen concentrations. After 8 weeks of dietary interventions, there was no significant difference in body mass in any of dietary conditions (p >.05 ). After 3 hr swim exercise, blood lactate level was higher compared to resting conditions in all groups, but it was returned to resting value after 1 hr rest (p <.05 ). Free fatty acid concentration was higher in all high fat fed groups (regardless of fatty acid composition )than CHOW consumed group. At rest, pro- & macro-glycogen concentration was not different from any of experimental groups (p >.05 ). Regardless of forms of glycogen, the highest level was observed in liver (p <.01 ), and most cases of supercompensation after 3hr exercise observed in this study were occurred in CHOW fed tissues. Except heart muscle, all tissues used in this study showed that pro- and macro-glycogen concentration was significantly decreased after 3 hr exercise. Based on these results, two conclusions were made :first, there is no different level of glycogen content in various tissues regardless of types of fatty acids consumed and second, the highest mobilization rate would be demon-strated from CHOW fed animals compare to animals that consumed any kinds of fatty acid diet if prolonged exercise is applied.
Animals
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Diet
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Fatty Acids
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Glycogen
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Heart
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Lactic Acid
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Liver
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Muscle, Skeletal*
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Myocardium
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Rats*
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Rats, Sprague-Dawley
2.in vivo multiplex gene targeting with Streptococcus pyogens and Campylobacter jejuni Cas9 for pancreatic cancer modeling in wildtype animal
Yoo Jin CHANG ; Jihyeon BAE ; Yang ZHAO ; Geonseong LEE ; Jeongpil HAN ; Yoon Hoo LEE ; Ok Jae KOO ; Sunmin SEO ; Yang-Kyu CHOI ; Su Cheong YEOM
Journal of Veterinary Science 2020;21(2):e26-
Pancreatic ductal adenocarcinoma is a lethal cancer type that is associated with multiple gene mutations in somatic cells. Genetically engineered mouse is hardly applicable for developing a pancreatic cancer model, and the xenograft model poses a limitation in the reflection of early stage pancreatic cancer. Thus, in vivo somatic cell gene engineering with clustered regularly interspaced short palindromic repeats is drawing increasing attention for generating an animal model of pancreatic cancer. In this study, we selectedKras, Trp53, Ink4a, Smad4, and Brca2 as target genes, and applied Campylobacter jejuni Cas9 (CjCas9) andStreptococcus pyogens Cas9 (SpCas9) for developing pancreatic cancer using adeno associated virus (AAV) transduction. After confirming multifocal and diffuse transduction of AAV2, we generated SpCas9 overexpression mice, which exhibited high double-strand DNA breakage (DSB) in target genes and pancreatic intraepithelial neoplasia (PanIN) lesions with two AAV transductions; however, wild-type (WT) mice with three AAV transductions did not develop PanIN. Furthermore, small-sized Cjcas9 was applied to WT mice with two AAV system, which, in addition, developed high extensive DSB and PanIN lesions. Histological changes and expression of cancer markers such as Ki67, cytokeratin, Mucin5a, alpha smooth muscle actin in duct and islet cells were observed. In addition, the study revealed several findings such as 1) multiple DSB potential of AAV-CjCas9, 2) peri-ductal lymphocyte infiltration, 3) multi-focal cancer marker expression, and 4) requirement of > 12 months for initiation of PanIN in AAV mediated targeting. In this study, we present a useful tool for in vivo cancer modeling that would be applicable for other disease models as well.