Background
Methods
Chemicals
Experimental animals and treatments
Total RNA isolation from the cerebral cortex
Expression profiling using Rat 230 2.0 Expression Arrays
Data analysis
Quantitative real-time PCR
Results
Expression profiling revealed numerous ERα agonist-regulated immunity genes in the middle-aged female neocortex
GENE | MICROARRAY | REAL-TIME PCR | |||
---|---|---|---|---|---|
Symbol | Name | probe set | FC | TaqMan ID | RQ |
S100a9
| S100 calcium binding protein A9 | 1387125_at | 3.655 | Rn00585879_m1 | 2.029 |
S100a8
| S100 calcium binding protein A8 | 1368494_at | 3.458 | Rn00587579_g1 | 2.558 |
RT1-Aw2
| RT1 class Ib | 1388202_at | 0.291 | Rn03034964_u1 | 0.658 |
RatNP-3b
| defensin ratNP-3 precursor | 13700791_at | 3.317 | Rn01478511_gH | 10.568 |
IgG-2a
| immonoglobulin G | 1370394_at | 3.204 | Rn01429839_g1 | 9.257 |
Np4
| defensin NP-4 precursor | 1370470_at | 3.031 | Rn00597762_g1 | 2.268 |
Defa
| defensin, alpha 5, Paneth cell-specific | 1387943_at | 2.732 | Rn02607254_g1 | 32.763 |
Prg2
| proteoglycan 2 | 1387633_at | 2.412 | Rn00581137_m1 | 1.257 |
Camp
| cathelicidin antimicrobial peptide | 1393603_at | 2.297 | Rn01446021_g1 | 1.521 |
Mcpt9
| mast cell protease 9 | 1368501_s_at | 2.235 | Rn00755366_g1 | 5.825 |
Fcgr2b
| Fc fragment of IgG, receptor | 1371079_at | 0.451 | Rn00598391_m1 | 0.649 |
Igh-1a
| immunoglobulin heavy chain | 1388272_at | 2.144 | ||
Cd74
| CD74 | 1367679_at | 0.473 | Rn00565062_m1 | 0.685 |
Fcnb
| ficolin beta | 1387378_at | 2.071 | Rn00586231_m1 | 1.922 |
Mcpt8
| mast cell protease 8 | 1369586_at | 2.056 | Rn01789238_g1 | 3.784 |
Igj
| immunoglobulin joining chain | 1383163_at | 1.945 | Rn01768305_m1 | 1.204 |
Igha
| immunoglobulin heavy chain, alpha | 1371262_at | 1.866 | ||
RT1-Aw2
| RT1 class Ib | 1388203_x_at | 0.536 | Rn03034964_u1 | 0.658 |
Serping1
| C1-Inhibitor | 1372254_at | 0.559 | Rn01485600_m1 | 0.690 |
C3
| complement C3 | 1368000_at | 0.559 | Rn00566466_m1 | 0.743 |
RT1-N1
| RT1 class Ib, locus N1 | 1387839_at | 0.570 | Rn00561858_m1 | 0.654 |
Lrrc8a
| leucine rich repeat containing 8 family | 1382920_at | 1.670 |
rank | pathway | set size | percent up | average (NTk, NEk) |
---|---|---|---|---|
1
|
graft-versus-host disease
|
67
|
9
|
7.2
|
2
|
autoimmune thyroid disease
|
73
|
8
|
7.5
|
3
|
allograft rejection
|
68
|
9
|
7.8
|
4 | cell adhesion molecules | 189 | 18 | 8.0 |
5 | retinol metabolism | 64 | 11 | 9.5 |
6 | type I diabetes mellitus | 77 | 16 | 11.2 |
7
|
hematopoietic cell lineage
|
101
|
18
|
13.5
|
8 | neuroactive ligand-receptor interaction | 335 | 16 | 13.8 |
9 | pantothenate and CoA biosynthesis | 11 | 27 | 14.5 |
10
|
C and coagulation cascades
|
92
|
13
|
14.5
|
11 | androgen and estrogen metabolism | 33 | 15 | 16.5 |
12 | Parkinson's disease | 139 | 69 | 16.5 |
13
|
cytokine-cytokine receptor interaction
|
210
|
23
|
20.5
|
14
|
systemic lupus erythematosus
|
92
|
22
|
22.2
|
15 | caffeine metabolism | 10 | 10 | 25.5 |
16 | metabolism of xenobiotics by cytochrome P450 | 48 | 15 | 26.2 |
17 | ECM-receptor interaction | 101 | 24 | 26.2 |
18
|
Jak-STAT signaling pathway
|
172
|
28
|
27.2
|
19 | drug metabolism - other enzymes | 38 | 24 | 27.5 |
20 | basal cell carcinoma | 68 | 21 | 29.0 |
Examination of the estrogenic regulation of neuroinflammatory genes
Genes regulated by E2
Symbol | Gene name | TaqMan ID | RQ(E2) | p |
---|---|---|---|---|
complement
| ||||
C3
| complement C3 | Rn00566466_m1 | 0.70 | 0.02 |
C4b
| complement C4 | Rn00709527_m1 | 0.67 | 0.07 |
defensin
| ||||
Np4
| defensin NP-4 precursor | Rn00597762_g1 | 2.56 | 0.08 |
RatNP-3b
| defensin ratNP-3 precursor | Rn01478511_gH | 13.27 | 0.06 |
Ig chain
| ||||
IgG-2a
| immonoglobulin G | Rn01429839_g1 | 8.23 | 0.10 |
lymphokine
| ||||
Ccl2
| chemokine (C-C) ligand 2 | Rn00580555_m1 | 0.52 | 0.06 |
Il6
| interleukin-6 | Rn00561420_m1 | 2.82 | 0.04 |
Tgfb1
| transforming growth factor beta 1 | Rn00572010_m1 | 0.78 | 0.01 |
MHC
| ||||
RT1-Aw2
| RT1 class Ib | Rn03034964_u1 | 0.61 | 0.10 |
miscellaneous
| ||||
Cx3cr1
| fractalkine receptor | Rn00591798_m1 | 0.82 | 0.04 |
Mpeg1
| macrophage expressed gene 1 | Rn02769865_s1 | 0.61 | 0.01 |
nuclear receptor
| ||||
Esr1
| estrogen receptor-α | Rn00562166_m1 | 1.39 | 0.05 |
phagocytic receptor
| ||||
Fcgr2b
| Fc fragment of IgG, receptor | Rn00598391_m1 | 0.61 | 0.03 |
Itgam
| CD11b | Rn00709342_m1 | 0.57 | 0.01 |
Toll-like receptor
| ||||
Tlr4
| Toll-like receptor 4 | Rn00569848_m1 | 0.81 | 0.05 |
Tlr9
| Toll-like receptor 9 | Rn01640054_m1 | 0.74 | 0.07 |
Neuroinflammatory genes regulated by both E2 and isotype selective ER agonists
Symbol | Gene name | TaqMan ID | RQ(LE2) | RQ(DPN) |
---|---|---|---|---|
Complement
| ||||
C3
| complement C3 | Rn00566466_m1 | 0.743 | 0.801 |
Defensin
| ||||
Np4
| defensin NP-4 precursor | Rn00597762_g1 | 2.268 | 2.578 |
RatNP-3b
| defensin ratNP-3 precursor | Rn01478511_gH | 10.568 | 4.574 |
Ig chain
| ||||
IgG-2a
| immonoglobulin G | Rn01429839_g1 | 9.257 | 4.048 |
Lymphokine
| ||||
Ccl2
| chemokine (C-C) ligand 2 | Rn00580555_m1 | 0.824 | 0.703 |
Il6
| interleukin-6 | Rn00561420_m1 | 2.232 | 2.635 |
MHC
| ||||
RT1-Aw2
| RT1 class Ib | Rn03034964_u1 | 0.658 | 0.463 |
phagocytic receptor
| ||||
Fcgr2b
| Fc fragment of IgG, receptor | Rn00598391_m1 | 0.649 | 0.651 |
Itgam
| CD11b | Rn00709342_m1 | 0.655 | 0.798 |
Symbol | Gene name | RQ(E2) | RQ(LE2) | RQ(DPN) |
---|---|---|---|---|
Analogous changes
| ||||
C3
| complement C3 | 0.703 | 0.743 | 0.801 |
Ccl2
| chemokine (C-C) ligand 2 | 0.527 | 0.824 | 0.703 |
Fcgr2b
| Fc fragment of IgG, receptor | 0.615 | 0.649 | 0.651 |
IgG-2a
| Ig chain | 8.228 | 9.257 | 4.048 |
Il6
| interleukin-6 | 2.823 | 2.232 | 2.635 |
Itgam
| CD11b | 0.571 | 0.655 | 0.798 |
Np4
| defensin NP-4 precursor | 2.562 | 2.268 | 2.578 |
RatNP-3b
| defensin ratNP-3 precursor | 13.266 | 10.568 | 4.574 |
RT1-Aw2
| RT1 class Ib | 0.606 | 0.658 | 0.463 |
E2-specific changes
| ||||
C4b
| C4 | 0.670 | 0.881 | 0.785 |
Cx3cr1
| fractalkine receptor | 0.820 | 0.914 | 1.041 |
Esr1
| estrogen receptor-α | 1.392 | 1.169 | 0.908 |
Mpeg1
| macrophage expressed gene 1 | 0.611 | 0.978 | 0.869 |
Tgfb1
| transforming growth factor beta 1 | 0.779 | 0.921 | 0.938 |
Tlr4
| Toll-like receptor 4 | 0.810 | 0.993 | 1.069 |
Tlr9
| Toll-like receptor 9 | 0.737 | 0.896 | 0.845 |