10/30/2008 Adipose Tissue • What impact does adipose tissue growth and development have on livestock production? p p – meat quality, economics – health concerns in consuming extra fat – calorie density‐obesity – source of hormones and signal molecules regulate body metabolism • Adi Adipose Tissue (A.T.) = connective tissue Ti (A T ) ti ti classification, consists of many cell types TTU AFS 4400/5400 1 Adipose Tissue • Meat Quality and economics Comparison with carcass grade among Korea, Japan, and USA TTU AFS 4400/5400 TTU AFS 4400/5500 2 1 10/30/2008 Comparison of Angus and American Wagyu beef Choice Angus Wagyu TTU AFS 4400/5400 3 Adipose Tissue • Health concerns in consuming extra fat The Beef Marbling Score (BMS) is directly related to the percentage extractable lipid. TTU AFS 4400/5400 TTU AFS 4400/5500 4 2 10/30/2008 Adipose Tissue • Health concerns in consuming extra fat Fatty acid composition of intramuscular adipose tissue (marbling) of U.S. crossbred cattle or Japanese Black cattle raised in Japan. (Adapted from Sturdivant et al., 1992.) TTU AFS 4400/5400 5 MUFA on cholesterol metabolism in human subjects • Health concerns in consuming extra fat • 10 men consumed ground beef fomulated with Angus or Wagyu fat trim. - Angus 1: MUFA:SFA ratio = 0.82 - Angus 2: MUFA:SFA ratio = 1.34 - Wagyu: MUFA:SFA ratio = 1.34 • High MUFA diet reduce LDL cholesterol in plasma (Smith et al., Unpublished) TTU AFS 4400/5400 TTU AFS 4400/5500 6 3 10/30/2008 Adipose Tissue Distribution of fat between the subcutaneous, intermuscular, intramuscular, and kidney fat in beef, lamb, and pork carcass. Livestock & Carcasses by Boggs et al., 1998 TTU AFS 4400/5400 7 Adipose Tissue • Functions of AT – Energy Storage Energy Storage = 2.25 x Gross energy as 2.25 x Gross energy as protein or CHO – Insulation – Protection of vital organs – Non‐shivering thermogenesis • Brown adipose tissue p • release of heat – newborn animals – cold adaptation – hibernation TTU AFS 4400/5400 TTU AFS 4400/5500 8 4 10/30/2008 Adipose Tissue • Four major depots of AT – visceral i l – intermuscular (“seam fat”) – subcutaneous (“backfat”) – intramuscular (“marbling”) TTU AFS 4400/5400 9 Adipose Tissue • Largest triglyceride in animal adipose tissue – Palmitodiolein ( Glycerol + palmitic, oleic, oleic acid) – Oleopalmitostearin (Glycerol + oleic, palmitic, stearic acid) – Oleic acid is largest portion in animal adipose tissue TTU AFS 4400/5400 TTU AFS 4400/5500 10 5 10/30/2008 Adipose Tissue • Adipose Tissue (A.T.) = connective tissue classification, consists of many cell types , y yp • Adipose tissue generally composed of: 76‐94% lipid 1‐4% protein 5‐20% water. TTU AFS 4400/5400 11 Adipose Tissue • Lipid or Fat ≠ AT – Lipid/Fat is just one portion of AT – adipocyte di t stores lipid/fat t li id/f t • Lipids = mainly triacylglycerols (TAGs) • Adipose Tissue Structure – eyelids and scrotum completely devoid of AT – largest “organ” in body – Human males AT = 15‐20% of body weight, females 20‐25% of body wt. – tremendous capacity to expand mainly because it is very vascular – vascularity related to # of cells at the specific AT deposition – metabolically active, based on energy balance of animal TTU AFS 4400/5400 TTU AFS 4400/5500 12 6 10/30/2008 Adipose Tissue • Types of lipids • Ruminant Animal contained cis-9 trans-11, trans-10, cis-12 conjugated linoleic acid (CLA) TTU AFS 4400/5400 13 Conjugated linoleic acid • trans-10, cis-12 CLA • cis-9, trans-11 CLA • linoleic acid J. Chem. Edu, 73:302–303, 1996. TTU AFS 4400/5400 TTU AFS 4400/5500 14 7 10/30/2008 Adipose Tissue • Biohydrogenation in ruminants. TTU AFS 4400/5400 15 Adipose Tissue SFA MUFA PUFA 80 70 P‐values *Endpoint : P < 0.01 Percentage 60 50 40 30 20 10 0 U.S. DIGESTA* Jpn U.S. PLASMA* Jpn U.S. FattyTTU AFS 4400/5400 acid composition TTU AFS 4400/5500 Jpn S.C. ADIPOSE TISSUE* 16 8 10/30/2008 Adipose Tissue Growth • The increase of adipose tissue ‐ Adipocyte Number + Adipocyte Number + Adipocyte Size. Size • The source of new fat cell ‐ Fibroblast, Fibroblast‐like cell ‐‐‐ Adipocyte • Brown Adipocyte ‐ precursor for white adipocyte precursor for white adipocyte ‐ No brown fat in piglet. • Net lipid accretion rate = rate of lipigenesis‐ rate of lipolysis TTU AFS 4400/5400 17 Adipose Tissue Growth • Adipose tissue : major lipogenesis in cattle, sheep pig sheep, pig • Liver : major lipogenesis in rodents, avian, human TTU AFS 4400/5400 TTU AFS 4400/5500 18 9 10/30/2008 Adipose Tissue Growth • Monogastric animal : Dietary FA used for body adipocyte triglyceride synthesis ‐ Glucose is major substrate for lipogenesis • Ruminants : Biohydrogenation changed dietary FA from unsaturated to saturated ‐ Acetate is major substrate for lipogenesis ‐ Relatively low ATP‐citrate lyase, NADP‐malate dehydrogenase in liver • Poultry : Used VLDL for transfer lipid in body TTU AFS 4400/5400 19 Stearic acid vs. slip point • Positive correlation. U.S. endpoint • Slip point was highest in hay‐fed steers. • U.S. endpoint steers were distributed in high Slip point and high y = ‐0.039x2 + 2.23x + 12.07 R2 = 0.917 P < 0.001 TTU AFS 4400/5500 18:0. 10 10/30/2008 Stearic acid vs. palmitoleic acid U.S. endpoint • Negative correlation. • 16:1n‐7 was highest in corn‐fed steers. • Japanese endpoint steers were distributed steers were distributed y = 0.015x2 ‐ 7.96x + 12.32 R2 = 0.925 P < 0.001 in high 16:1 and low 18:0. Percentage intramuscular lipid as a function of slaughter age. Abbreviations for breed types: B J B, Japanese Black; Bl k BHo, J. Black x Holstein; BBHo, J. Black x J.Black/Holstein; RBHo, J. Brown x J.Black/Holstein; CBHo, Charolais x J. Black/Holstein; NBHo, J. Shorthorn x J. Black/Holstein. / All cattle were fed a high concentrate finishing diet. Zembayashi et al. (1999). TTU AFS 4400/5500 11 10/30/2008 Diet effects on lipogenesis • High-energy High energy diet increased lipogenic enzyme activities (Prior and Scott, 1980). • Corn diet increased ACC, FAS, ATP-citrate lyase, and NADP-malate dehydrogenase activities ((Smith et al.,, 1984). ) • Acetate is used as major carbon source in s.c. adipose tissue (Smith and Crouse, 1984). Diet effects on lipogenesis • Glucose is used as major carbon source in i.m. im adipose tissue (Smith and Crouse, 1984). • Corn-based diets increase bypass ratio. • Corn-based diets increase propionate production in rumen. TTU AFS 4400/5500 12 10/30/2008 Adipose Tissue • Fatty acid synthesis 1. De novo synthesis : synthesis from non-fatty acid precursors a) b) c) Carbohydrate precursors (glucose, lactate, and pyruvate) Amino acid precursors (alanine, branched-chain amino acids) Short-chain organic acids (acetate, propionate) 2 Lipogenesis : fatty acid or triacylglycerol synthesis 2. a) b) From preformed fatty acids (from diet or de novo fatty acid synthesis) Requires source of carbon for glycerol backbone TTU AFS 4400/5400 25 Adipose Tissue Glucose Lactate Acetate Pyruvate Acetyl-CoA Fructose1.6-bisP GAP Mitochondria TTU AFS 4400/5400 TTU AFS 4400/5500 Fatty acid synthesis Citrate Lipogenesis in cattle 26 13 10/30/2008 Adipose Tissue Acetate ACC Acetyl-CoA FAS Malonyl-CoA Butyryl-CoA Palmitate 16:0 Elongation SCD Stearate 18:0 16 : 1 SCD 18 : 1 Fatty acid synthesis TTU AFS 4400/5400 27 Function of Fatty Acid • Role of fatty acid in Adipose tissue. Mit h d i Mitochondria UCP Lipid Oxidation -Energy expenditure Heat increase RXR Adipocyte Nucleus PPARγ H+ Transcriptional Factor -DNA binding protein Fatty Acids = O mRNA Energy Storage -Triacylglycerol Droplet = DNA CH2-O-C-(CH2)n-CH3 O Membrane molecules -Phospholipids Phospholipids = CH2-O-C-(CH2)n-CH3 O CH2-O-C-(CH2)n-CH3 TTU AFS 4400/5400 TTU AFS 4400/5500 28 14 10/30/2008 Adipose Tissue • Types of Adipose Tissue – White adipose tissue (WAT) • signet ring, mature cell surrounded by clear epithelial layer • larger than brown (35‐200 μm) • nucleus pushed to the outside by the lipid droplet TTU AFS 4400/5400 29 Adipose Tissue – Brown adipose tissue (BAT) • • • • • small cells (20‐25 μm) ll ll (20 25 ) much more connective tissue gland‐like appearance many mitochondria uncoupling proteins (UCP‐1) TTU AFS 4400/5400 TTU AFS 4400/5500 30 15 10/30/2008 Adipocyte Adipocytes = primary cell of AT; AT also contains other connective tissue vascular tissue and other connective tissue, vascular tissue, and nerve cells TTU AFS 4400/5400 31 Preadipocyte Differentiation • Preadipocyte Proliferation/Differentiation – cell line: 3T3‐L1 Æ once differentiation begins, proliferation ceases – early marker of differentiation of 3T3‐L1 is SCD (stearoyl coenzyme A desaturase) TTU AFS 4400/5400 TTU AFS 4400/5500 32 16 10/30/2008 Adipocyte • Differentiation of Adipocytes – adipocytes are major TG‐containing components of adipose tissue (85‐95% of cell type) – adipocytes are differentiated from mesenchymal precursors – adipoblasts Æ preadipocytes (in utero) • both undergo cell division – preadipocytes are immature adipocytes that can still proliferate – hard to isolate preadipocytes in embryonic development – Thus, the majority of research has utilized a secondary cell line (3T3‐ Th th j it f hh tili d d ll li (3T3 L1; preadipocytes) derived from murine fibroblasts TTU AFS 4400/5400 33 Morphology of Adipocyte • Perirenal adipocytes • Proliferation: 10% fetal bovine serum + DMEM • Differentiation: 5% fetal bovine serum + DMEM + insulin + Pioglitazone + MIX • 15 d after treatment with differentiation media TTU AFS 4400/5400 TTU AFS 4400/5500 34 17