Table 1.
Relative gene expression for osteoclast-associated genes, chemokines, and chemokine receptors
|
Expression, microarray relative units ± 1 SD
|
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|---|---|---|---|---|---|---|---|
| Gene name | Alternative names | GenBank accession no. | RAE230 array probe set | Day 0 | Day 2 | Day 4 | Day 6 |
| Control/osteoclast genes | |||||||
| GAPDH | — | NM_017008 | 1367557_s_at | 9750 ± 2181 | 10 737 ± 1 446 | 11 282 ± 2 216 | 11 111 ± 1 496 |
| Acid phosphatase 5 | TRAP | NM_019144 | 1367942_at | 1212 ± 378 | 6 677 ± 2 294 | 12 001 ± 739 | 10 454 ± 265 |
| Cathepsin K | — | NM_031560 | 1369947_at | 1789 ± 487 | 8 906 ± 2 818 | 14 564 ± 994 | 12 657 ± 1 475 |
| RANKL | TRANCE, TNF superfamily member 11 | NM_057149 | 1369282_at | 679 ± 209 | 981 ± 263 | 484 ± 117 | 385 ± 59 |
| OPG | Osteoprotegerin, TNF receptor superfamily, member 11b | NM_012870 | 1369407_at | 325 ± 54 | 331 ± 15 | 452 ± 75 | 422 ± 63 |
| CSF-1 receptor | c-Fms | BI285793 | 1388784_at | 770 ± 99 | 4 310 ± 1 337 | 3 127 ± 644 | 2 462 ± 327 |
| Chemokines/Receptors | |||||||
| CC chemokines | |||||||
| CCL1 (NRS) | — | — | — | — | — | — | — |
| CCL2 | MCP-1 | NM_031530 | 1367973_at | 189 ± 80 | 456 ± 184 | 534 ± 361 | 257 ± 77 |
| CCL3 | MIP-1α | U22414 | 1369815_at | 420 ± 94 | 566 ± 46 | 425 ± 70 | 429 ± 63 |
| CCL4 | MIP-1β | NM_053858 | 1370832_at | 385 ± 103 | 386 ± 34 | 323 ± 42 | 321 ± 36 |
| CCL5 | RANTES | NM_031116 | 1369983_at | 565 ± 83 | 450 ± 74 | 532 ± 112 | 545 ± 120 |
| CCL6 | MRP-1α | BE095824 | 1389123_at | 1131 ± 378 | 2 244 ± 720 | 919 ± 247 | 1 358 ± 367 |
| CCL7 | MCP-3 | BF419899 | 1379935_at | 117 ± 20 | 113 ± 10 | 128 ± 24 | 93 ± 26 |
| CCL8 (NRS) | MCP-2 | — | — | — | — | — | — |
| CCL9 | MIP-1γ | AI169984 | 1392172_at | 100 ± 94 | 7 832 ± 2 260 | 1 898 ± 783 | 858 ± 385 |
| CCL11 | Eotaxin | NM_019205 | 1387319_at | 257 ± 20 | 243 ± 26 | 218 ± 29 | 213 ± 18 |
| CCL12 (NP) | MCP-5 | XM_213425 | — | — | — | — | — |
| CCL13 (NRS) | MCP-4 | — | — | — | — | — | — |
| CCL15 (NRS) | HCC-2 | — | — | — | — | — | — |
| CCL17 | TARC | NM_057151 | 1370118_at | 187 ± 11 | 175 ± 8 | 166 ± 12 | 185 ± 26 |
| CCL19 | ELC, MIP-3β | AA996885 | 1391925_at | 622 ± 195 | 478 ± 144 | 517 ± 108 | 517 ± 196 |
| CCL20 | MIP-3α | AF053312 | 1369814_at | 229 ± 23 | 196 ± 22 | 223 ± 13 | 274 ± 50 |
| CCL21 | 6CKINE | BI282920 | 1378015_at | 320 ± 26 | 417 ± 56 | 374 ± 104 | 431 ± 49 |
| CCL22 | MDC | AF432871 | 1370619_at | 207 ± 12 | 208 ± 23 | 178 ± 34 | 169 ± 44 |
| CCL25 (NP) | TECK | — | — | — | — | — | — |
| CCL26 | Eotaxin-3 | BI294084 | 1385309_at | 662 ± 181 | 742 ± 67 | 804 ± 145 | 636 ± 64 |
| CCL27 | CTACK | XM_342823 | 1376850_a_at | 463 ± 27 | 390 ± 40 | 356 ± 66 | 386 ± 84 |
| CXC chemokines | |||||||
| CXCL1 | GROα | NM_030845 | 1387316_at | 96 ± 12 | 115 ± 6 | 123 ± 21 | 108 ± 34 |
| CXCL2 | GROβ, MIP-2α | NM_053647 | 1368760_at | 498 ± 124 | 406 ± 72 | 408 ± 47 | 362 ± 172 |
| CXCL4 | Platelet factor 4, PF-4 | AI169104 | 1371 250_at | 5004 ± 632 | 5 172 ± 297 | 7 687 ± 566 | 10 210 ± 538 |
| CXCL5 | CXC chemokine LIX, ENA-78 | NM_022214 | 1387648_at | 233 ± 100 | 148 ± 148 | 156 ± 41 | 119 ± 72 |
| CXCL8 (NRS) | IL-8 | — | — | — | — | — | — |
| CXCL9 | MIG | AI044222 | 1382454_at | 155 ± 19 | 137 ± 12 | 142 ± 30 | 137 ± 15 |
| CXCL10 | CRG-2, IP-10 | U22520 | 1387969_at | 173 ± 20 | 152 ± 29 | 171 ± 18 | 190 ± 29 |
| CXCL11 | I-TAC | BF281987 | 1379365_at | 90 ± 21 | 90 ± 15 | 86 ± 12 | 94 ± 19 |
| CXCL12 | SDF-1 | AI171777 | 1369633_at | 4661 ± 1007 | 5 563 ± 1 201 | 5 371 ± 917 | 5 893 ± 1 062 |
| CXCL13 | BCA-1 | AA892854 | 1398390_at | 353 ± 54 | 351 ± 54 | 359 ± 66 | 421 ± 129 |
| CXCL14 | BRAK, BOLOKINE | AI412979 | 1396129_at | 505 ± 69 | 838 ± 440 | 493 ± 106 | 675 ± 101 |
| XCL1 | Lymphotactin | NM_134361 | 1387831_at | 53 ± 27 | 36 ± 44 | 63 ± 62 | 64 ± 30 |
| CX3CL1 | Fraktalkine | NM_134455 | 1368200_at | 228 ± 112 | 257 ± 29 | 215 ± 69 | 239 ± 53 |
| Receptors | |||||||
| CCR1 | MIP1α-R, RANTES-R | NM_020542 | 1370083_at | 237 ± 74 | 1 122 ± 310 | 829 ± 306 | 523 ± 117 |
| CCR2 | MCP-1 R | NM_021866 | 1387742_at | 135 ± 45 | 155 ± 27 | 125 ± 9 | 115 ± 38 |
| CCR3 | Eotaxin R | NM_053958 | 1369808_at | 297 ± 97 | 244 ± 67 | 224 ± 21 | 223 ± 63 |
| CCR4 | — | NM_133532 | 1369555_at | 189 ± 49 | 187 ± 39 | 156 ± 19 | 185 ± 68 |
| CCR5 | — | NM_053960 | 1369290_at | 115 ± 18 | 126 ± 23 | 107 ± 17 | 90 ± 14 |
| CCR6 | GRO/MGSA-R, STRL22 | AI045155 | 1393929_at | 1148 ± 126 | 1 075 ± 142 | 1 180 ± 122 | 1 031 ± 168 |
| CCR7 (NP) | — | — | — | — | — | — | — |
| CCR8 (NP) | — | — | — | — | — | — | — |
| CCR9 (NP) | — | — | — | — | — | — | — |
| CCR10 (NP) | — | — | — | — | — | — | — |
| CXCR1 | IL-8 RA | NM_019310 | 1369875_at | 288 ± 33 | 259 ± 52 | 272 ± 30 | 270 ± 29 |
| CXCR2 | IL-8 RB | NM_017183 | 1369697_at | 241 ± 37 | 223 ± 20 | 260 ± 31 | 242 ± 50 |
| CXCR3 | MIG-R, IP-10 | NM_053415 | 1368192_at | 312 ± 46 | 312 ± 56 | 356 ± 69 | 318 ± 56 |
| CXCR4 | LCR1, SDF1-R, Fusin | U54791 | 1389244_x_at | 4083 ± 521 | 4 902 ± 110 | 3 203 ± 614 | 4 156 ± 619 |
| CXCR5 | Burkitt lymphoma receptor 1 | NM_053303 | 1369911_at | 669 ± 104 | 513 ± 65 | 524 ± 106 | 507 ± 63 |
| CXCR6 (NRS) | — | — | — | — | — | — | — |
| CX3CR1 | V28 | NM_133534 | 1369527_at | 520 ± 96 | 411 ± 43 | 468 ± 96 | 508 ± 94 |
| XCR1 (NP) | GPR-5 | — | — | — | — | — | — |
NRS indicates no sequence for rat in GenBank with sufficient homology to assign identity; NP, no sequence present on the RAE 230 chipset used. mRNA levels were determined by DNA microarray analysis of tibial RNA isolated from toothless (tl/tl) rats on day 0 (3 weeks old), or following 2, 4, and 6 days of CSF-1 injections to induce osteoclast differentiation. Values shown are the mean ± 1 SD of 4 individual animals. GAPDH served as an internal standard. Osteoclast-specific genes include TRAP, cathepsin K, and the CSF-1 receptor (c-Fms). We also show results for the osteoblast-derived osteoclast-inducing factor RANKL and its decoy receptor OPG, which underwent little or no change in this experimental system. TRAP and cathepsin K mRNA levels follow the same time course as osteoclast counts in bone sections. CCL9 and its receptor CCR1 showed by far the greatest responses of all chemokines and receptors and peaked a full 2 days before osteoclast counts, as did CSF-1 receptor levels. Chemokine and receptor identities were assigned based on highest homology to mouse, determined as described in “Results.”