Post on 11-Apr-2017
MCP-1 and MIP-3α Expression in LCR/HCR RatsDr. Anuska Andjelkovic Lab By: Zarion Marshall
Low Capacity Runners (LCR) and High Capacity Runners (HCR) Rats
• LCR rats are highly susceptible to cardiovascular diseases like atherosclerosis
• LCR rats are insulin resistant
• Natural screening model (no gene manipulation)
• Grouped by simple measure of treadmill running distance
My Rats
• Generation 23• About 46-48 weeks old (~10-11 months)• All female for lower variability of body weight
LCR HCR0.00
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LCR HCR0.00
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LCR more sensitive to harmful effects of high fat diet
• Overall high body weight
• Higher fat percentage
• Higher food intake
• Higher serum leptin levels (mostly adipose)
Credit: Colleen M. Novak, Carlos Escande, Paul R. Burghardt, Minzhi Zhang, Maria Teresa Barbosa, Eduardo N. Chini, Steven L. Britton, Lauren G. Koch, Huda Akil, James A. Levine in “Spontaneous activity, economy of activity, and resistance to diet-induced obesity in rats bred for high intrinsic aerobic capacity”
Activity and Energy Expenditure
• LCR less active (not dependent on body weight) and expend less energy
Credit: Colleen M. Novak, Carlos Escande, Paul R. Burghardt, Minzhi Zhang, Maria Teresa Barbosa, Eduardo N. Chini, Steven L. Britton, Lauren G. Koch, Huda Akil, James A. Levine in “Spontaneous activity, economy of activity, and resistance to diet-induced obesity in rats bred for high intrinsic aerobic capacity”
Energy Expenditure and Respiratory Exchange Rate (RER)• LCR lower
energy expenditure even in rest
• Also higher RER which suggests ineffecient use of lipids
Credit: Colleen M. Novak, Carlos Escande, Paul R. Burghardt, Minzhi Zhang, Maria Teresa Barbosa, Eduardo N. Chini, Steven L. Britton, Lauren G. Koch, Huda Akil, James A. Levine in “Spontaneous activity, economy of activity, and resistance to diet-induced obesity in rats bred for high intrinsic aerobic capacity”
Hypothesis• LCR rats are most susceptible to proinflammatory
factors which leaves the blood brain barrier (BBB) at risk for microvascular changes like stroke
• Chemokines on basolateral (brain) side of barrier attract leukocytes from the apical (blood) side
Goal• Investigate the chemokines monocyte
chemotactic protein (MCP-1) and macrophage inflammatory protein (MIP-3α) and their role in the BBB’s integrity
Blood Brain Barrier
How It Works (Supposedly)
• Proinflammatory proteins cause barrier to lose integrity between endothelial cells
Protein Array (Vessels)
Leptin 349.67 63.58 0.181829
LIX 308.76 984.77 3.189435
L-Selectin 192.28 603.06 3.136364
MCP-1 311.31 1,782.95 5.727249
MIP-3α 39.78 779.87 19.60458
MMP-8 144.55 383.52 2.6532
PDGF-AA 4,323.15 3,028.13 0.700445
Prolactin R 4,737.59 5,581.02 1.178029
RAGE 3,582.80 6,503.18 1.815111
Thymus Chemokine-1 4,596.71 7,274.25 1.582491
TIMP-1 178.61 982.7 5.501932
HCR
LCR
HCR
LCR
Cytokine Expression
Activi
n A
B7-2/
CD86CIN
C-1CIN
C-3
Fas Lig
and
GM-CSF
IFN-ga
mma
IL-1bet
a IL-2 IL-6 IL-13 LIX
MCP-1MMP-8
Prolac
tin R
Thymus
Chem
okine
-1
TNF-a
lpha
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HCRLCR
Inflammatory Mediators
Rela
tive
Expr
essi
on
MCP-1(Tissue ELISA)
• 0.03 ng/ml HCR• 1.12 ng/ml LCR
HCR LCR0
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1.2
MCP-1 in Brain Tissue
Conc
entr
atio
n (n
g/m
l)
MIP-3α (Tissue ELISA)
• 0.14 pg/ml HCR• 2.72 pg/ml LCR
HCR LCR0
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MIP-3α in Brain Tissue
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entr
atio
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Staining
• Red=chemokine • Green=astrocyte• MIP-3α seems to be
mostly in astrocytes • You can see from the
staining as well not as much of either in HCR
• We think MCP-1 is more in microglia…
LCR HCR
MCP
-1M
IP-3
α
Conclusion• The higher inflammatory risk of LCR rats explains
at least in part their phenotype which resembles classic metabolic syndrome conditions
• MCP-1 and MIP-3α contribute are indeed factors that differentiate the two LCR and HCR phenotypes and suggests that a role in microvascular changes
Much Thanks to…• Svetlana and Nikola • And of course Dr. Andjelkovic