Adiponectin system in dairy cows during lactation (CLA effects and physiological changes) Behnam Saremi Institute of Animal Science, Physiology and Hygiene Unit, University of Bonn Munich 15.12.2010 Behnam Saremi

Introduction Adiponectin (Adi): involved in regulation of fat metabolism and insulin sensitivity and glucose homeostasis Adiponectin receptor 1 (AdiR1): ubiquitously expressed, particularly in skeletal muscle Adiponectin receptor 2 (AdiR2): high expression in liver tissue in comparison to other tissues (Yamauchi et al. 2005, Fu et al. 2005) Munich 15.12.2010 Behnam Saremi

Introduction It’s known from human and rodent researches that CLAs are functional signaling molecules interconnected to adipokine release in adipose tissue (AT). (Marshall, 2006) Munich 15.12.2010 Behnam Saremi

Objectives Therefore, the aim of this study was to investigate: Adiponectin and its receptors in different tissues specially fat depots of dairy cows in early to mid lactation CLA effects Physiological changes Munich 15.12.2010 Behnam Saremi

Days in milk and days of slaughter Materials and methods Heifers Treatments Days in milk and days of slaughter 1 42 105 Animals/ treatment Control 5 100 g CLA / animal / day - 6(5)* Data of one cow was removed because of disease therefore the actual number is superscripted. Munich 15.12.2010 Behnam Saremi

Samples Fat tissues: Subcutaneous(Sc): Tail head, Wither, Sternum Visceral(Vc): Retroperitoneal, Mesenterial, Omental Non fat tissues: Liver, Muscle, and Pancreas Munich 15.12.2010 Behnam Saremi

Munich 15.12.2010 Behnam Saremi

Cq: Quantification cycle value, RQ: relative quantity, NRQ: Normalized relative quantity, CNRQ: calibrated NRQ (Hellemans et al, 2007). Munich 15.12.2010 Behnam Saremi

mRNA abundance Data normalization based on geomean of at least three most stable reference genes (Vandesompele et al. 2002) qBaseplus software used for all calculations, data quality control and real time PCR efficiency correction (Hellemans et al. 2007) Statistical analysis: GLM or non parametric tests (p<0.05) SPSS 17 In most of the cases, it was 6 ref genes (in muscle and pancreas 4 and 5 in liver) Munich 15.12.2010 Behnam Saremi

Results Individual fat depots Vc and Sc fat depots Non fat tissues Munich 15.12.2010 Behnam Saremi

Days after parturition Adi mRNA abundance (Mean ± SEM) in Vc and Sc fat tissues of dairy cows fed with or without CLA Mesenterial Omental Retroperitoneal c CLA * b * b * a a a a mRNA abundance (A.U.) Sc Sternum Sc Tail head Sc Wither Days after parturition Munich 15.12.2010 Behnam Saremi

Days after parturition AdiR1 mRNA abundance (Mean ± SEM) in Vc and Sc fat tissues of dairy cows fed with or without CLA Mesenterial Omental Retroperitoneal c CLA * * * b a ab mRNA abundance (A.U.) Sc Sternum Sc Tail head Sc Wither * * * b b b a a a ab a Days after parturition Munich 15.12.2010 Behnam Saremi 12

Days after parturition AdiR2 mRNA abundance (Mean ± SEM) in Vc and Sc fat tissues of dairy cows fed with or without CLA Mesenterial Omental Retroperitoneal * c CLA b * a ab mRNA abundance (A.U.) Sc Sternum Sc Tail head Sc Wither Days after parturition Munich 15.12.2010 Behnam Saremi 13

Results Individual fat depots Vc and Sc fat depots Non fat tissues Munich 15.12.2010 Behnam Saremi

Days after parturition Adi mRNA abundance (Mean ± SEM) in Sc and Vc fat tissues of dairy cows in control group * * mRNA abundance (A.U.) Based on 6 ref genes Days after parturition Munich 15.12.2010 Behnam Saremi

Days after parturition AdiR1 mRNA abundance (Mean ± SEM) in Sc and Vc fat tissues of dairy cows in control group origin : p<0.003 * * mRNA abundance (A.U.) Based on 6 ref genes Main effect origin : p<0.003 Main effect origin : p<0.015 origin : p<0.015 Days after parturition Munich 15.12.2010 Behnam Saremi

Days after parturition AdiR2 mRNA abundance (Mean ± SEM) in Sc and Vc fat tissues of dairy cows in control group mRNA abundance (A.U.) Based on 6 ref genes Days after parturition Munich 15.12.2010 Behnam Saremi

Days after parturition Days after parturition Adi, AdiR1, and AdiR2 mRNA abundance (Mean ± SEM) in Sc fat tissues of dairy cows fed with or without CLA Adi mRNA abundance (A.U.) AdiR1 AdiR2 * * Days after parturition Days after parturition Munich 15.12.2010 Behnam Saremi

Days after parturition Days after parturition Adi, AdiR1, and AdiR2 mRNA abundance (Mean ± SEM) in Vc fat tissues of dairy cows fed with or without CLA Adi * * * mRNA abundance (A.U.) AdiR1 AdiR2 * p<0.067 * Days after parturition Days after parturition Munich 15.12.2010 Behnam Saremi 19

Adi mRNA abundance (Mean ± SEM) in individual fat tissues of dairy cows mRNA abundance (A.U.) b ab ab ab a a Based on 6 ref genes (Main effect of tissue) (All days together) Munich 15.12.2010 Behnam Saremi 20 20

AdiR1 mRNA abundance (Mean ± SEM) in individual fat depots of dairy cows mRNA abundance (A.U.) bc c b b b a Based on 6 ref genes (Main effect of tissue) (All days together) Munich 15.12.2010 Behnam Saremi 21 21

AdiR2 mRNA abundance (Mean ± SEM) in individual fat depots of dairy cows P<0.068 mRNA abundance (A.U.) Based on 6 ref genes (Main effect of tissue) (All days together) Treat*day:P<0.042 Munich 15.12.2010 Behnam Saremi 22 22

Results Individual fat depots Vc and Sc fat depots Non fat tissues Munich 15.12.2010 Behnam Saremi

Days after parturition AdiR1 mRNA abundance (Mean ± SEM) in liver and Muscle tissue of dairy cows fed with or without CLA Treatment: p<0.055 Liver Muscle * * a * mRNA abundance (A.U.) b b Days after parturition Munich 15.12.2010 Behnam Saremi

Days after parturition AdiR2 mRNA abundance (Mean ± SEM) in liver and Muscle tissue of dairy cows fed with or without CLA Treatment: p<0.098 Liver Muscle mRNA abundance (A.U.) Based on 5 ref genes Days after parturition Munich 15.12.2010 Behnam Saremi

Days after parturition AdiR1 mRNA abundance (Mean ± SEM) in pancreas tissue of dairy cows fed with or without CLA b b * b a a mRNA abundance (A.U.) Based on 4 ref genes Days after parturition Treat*day: p<0.049 Munich 15.12.2010 Behnam Saremi

Days after parturition AdiR2 mRNA abundance (Mean ± SEM) in pancreas tissue of dairy cows fed with or without CLA mRNA abundance (A.U.) Based on 4 ref genes Days after parturition Munich 15.12.2010 Behnam Saremi

Summary of results Adiponectin: Effect of day and treatment in VcATs AdR1: Day effect in VcATs, ScATs, liver, and pancreas AdR2: Day and treatment effect in retroperitoneal AT AdR2 mRNA abundance is not regulated by CLA and DIM except in retroperitoneal which a rose of AdR2 and decline by CLA in 105 DIM was detectable. (Numerically observed for Sc tail head) Munich 15.12.2010 Behnam Saremi

Summary of results AdR1: Sc>Vc In VcAT: Ad and AdR2 reduced by CLA in 105 DIM mRNA abundance of AdR2>AdR1 in all tissues Level of expression of adiponectin system is fat depots dependent AdR2 mRNA abundance is not regulated by CLA and DIM except in retroperitoneal which a rose of AdR2 and decline by CLA in 105 DIM was detectable. (Numerically observed for Sc tail head) Munich 15.12.2010 Behnam Saremi

Conclusion Because of the adiponectin role in insulin sensitivity, the reduction of adiponectin by CLA in 105 DIM could lead to lower insulin sensitivity in VcAT. The reaction of adiponectin system is different in Sc and VcAT regarding to CLA. The pattern of adiponectin system is comparable to the performance experiment regarding to no CLA effect at Sc tail head and liver and time effects in both tissues. Munich 15.12.2010 Behnam Saremi

Haptoglobin in dairy cows during lactation (CLA effects and physiological changes) Munich 15.12.2010 Behnam Saremi 31 31

Introduction Haptoglobin (Hp) is the most relevant acute phase protein in cattle, secreted mainly by the liver. In rats: supplementation with CLA affects Hp in serum, but not Hp mRNA abundance in liver. In dairy cows: In liver, mRNA expression of Hp is known. In adipose tissue (AT), less data were available until now. Munich 15.12.2010 Behnam Saremi 32

Objectives Therefore, the aim of this study was to investigate: Haptoglobin mRNA abundance in different tissues specially fat depots of dairy cows in early to mid lactation CLA effects Physiological changes Munich 15.12.2010 Behnam Saremi 33

Materials and methods Similar to previous Munich 15.12.2010 Behnam Saremi 34

Days after parturition Haptoglobin mRNA abundance (Mean ± SEM) in liver tissue of dairy cows fed with or without CLA a * * mRNA abundance (A.U.) b b Days after parturition Munich 15.12.2010 Behnam Saremi 35

Days after parturition Haptoglobin mRNA abundance (Mean ± SEM) in fat tissues of dairy cows fed with or without CLA CLA : p<0.05 * mRNA abundance (A.U.) CLA : p<0.05 Days after parturition Munich 15.12.2010 Behnam Saremi 36

Days after parturition Haptoglobin mRNA abundance (Mean ± SEM) in Sc and Vc fat tissues of dairy cows in control group Origin : p<0.05 * * mRNA abundance (A.U.) Based on 6 ref genes Days after parturition Munich 15.12.2010 Behnam Saremi 37 37

Correlations between Haptoglobin mRNA abundance and Kg and %EBW of fat tissues of dairy cows Munich 15.12.2010 Behnam Saremi 38

Thank you For your attention Munich 15.12.2010 Behnam Saremi