1. 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
Behnam Saremi

2. 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
Behnam Saremi

3. 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
Behnam Saremi

4. 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
Behnam Saremi

5. 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
Behnam Saremi

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

7. Munich
Behnam Saremi

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

9. 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
Behnam Saremi

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

11. 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
Behnam Saremi

12. 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
Behnam Saremi 12

13. 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
Behnam Saremi 13

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

15. 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
Behnam Saremi

16. 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< Main effect origin : p<0.015
origin : p<0.015
Days after parturition
Munich
Behnam Saremi

17. 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
Behnam Saremi

18. 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
Behnam Saremi

19. 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
Behnam Saremi 19

20. 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
Behnam Saremi 20 20

21. 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
Behnam Saremi 21 21

22. 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
Behnam Saremi 22 22

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

24. 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
Behnam Saremi

25. 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
Behnam Saremi

26. 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
Behnam Saremi

27. 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
Behnam Saremi

28. 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
Behnam Saremi

29. 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
Behnam Saremi

30. 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
Behnam Saremi

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

32. 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
Behnam Saremi 32

33. 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
Behnam Saremi 33

34. Materials and methods Similar to previous Munich 15.12.2010
Behnam Saremi 34

35. 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
Behnam Saremi 35

36. 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
Behnam Saremi 36

37. 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
Behnam Saremi 37 37

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

39. Thank you
For your attention
Munich
Behnam Saremi