Hippocampus Consortium M430v2 (Oct05) PDNN

Accession number: GN88

    Summary:

PRELIMINARY: The October 2005 Hippocampus Consortium data set provides estimates of mRNA expression in the adult hippocampus of approximately 96 genetically diverse strains of mice including 65 BXD recombinant inbred strains, 13 CXB recombinant inbred strains, a set of 16 diverse inbred strains, and 2 reciprocal F1 hybrids. The hippocampus is an important and intriguing part of the forebrain that is crucial for memory formation, and that is often affected in epilepsy, Alzheimer's disease, and schizophrenia. Unlike most other parts of the brain, the hippocampus contains a remarkable population of stems cells that continue to generate neurons and glial cells even in adult mammals (Kempermann, 2005). This genetic analysis of transcript expression in the hippocampus (dentate gyrus, CA1-CA3, and parts of the subiculum) is a joint effort of 14 investigators that is supported by numerous agencies described in the acknowledgments section. Samples were processed using a total of 206 Affymetrix Mouse Expression 430 2.0 short oligomer microarrays (MOE430 2.0 or M430v2), of which 178 passed stringent quality control and error checking. This particular data set was processed using the position-dependent nearest neighbor method (PDNN) of Zhang and colleagues. To simplify comparison among the transforms we have used, the quantile normalized PDNN values from each arrray have been adjusted to an average expression of 8 units and a standard deviation of 2 units.

    About the strains used to generate this set of data:

This analysis has used 65 of BXD strains, the complete set of 13 CXB recombinant inbred strain sets, and a mouse diversity panel consisting of 16 inbred strains and a pair of reciprocal F1 hybrids (B6D2F1 and D2B6F1).

The BXD genetic reference population of recombinant inbred strains consists of approximately 80 strains. Approximately 800 classical phenotypes from sets of 10 to 70 of these strains been integrated in the GeneNetwork. The BXD strains in this data set include 29 of the BXD strains made by Benjamin Taylor at the Jackson Laboratory in the 1970s and 1990s (BXD1 through BXD40). All of these strains are fully inbred, many well beyond the 100th filial (F) generation of inbreeding. We have also included 39 inbred (25 strains at F20+) and nearly inbred (14 strains between F14 and F20) BXD lines generated by Lu and Peirce. All of these strains, including those between F14 and F20, have been genotyped at 13,377 SNPs.

The CXB is the first and oldest set of recombinant inbred strains. Over 500 classical phenotypes from these strains been integrated in the GeneNetwork. It is noteworthy that the CXB strains segregate for the hippocampal lamination defect (Hld),characterized by Nowakowski and colleagues (1984). All of the CXBs have been recently genotyped at 13,377 SNPs.

Mouse Diversity Panel (MDP). We have profiled a MDP consisting inbred strains and a pair of reciprocal F1 hybrids; B6D2F1 and D2B6F1. These strains were selected for several reasons:

• genetic and phenotypic diversity, including use by the Phenome Project
• their use in making genetic reference populations including recombinant inbred strains, cosomic strains, congenic and recombinant congenic strains
• their use by the Complex Trait Consortium to make the Collaborative Cross (Nairobi/Wellcome, Oak Ridge/DOE, and Perth/UWA)
• genome sequence data from three sources (NHGRI, Celera, and Perlegen-NIEHS)
• availability from The Jackson Laboratory

All eight parents of the Collaborative Cross (129, A, C57BL/6J, CAST, NOD, NZO, PWK, and WSB) have been included in the MDP (noted below in the list). Twelve MDP strains have been sequenced, or are currently being resequenced by Perlegen for the NIEHS. This panel will be extremely helpful in systems genetic analysis of a wide variety of traits, and will be a powerful adjunct in fine mapping modulators using what is essentially an association analysis of sequence variants.

1. 129S1/SvImJ
       Collaborative Cross strain sequenced by NIEHS; background for many knockouts; Phenome Project A list
2. A/J
       Collaborative Cross strain sequenced by Perlegen/NIEHS; parent of the AXB/BXA panel
3. AKR/J
       Sequenced by NIEHS; Phenome Project B list
4. BALB/cJ
       Widely used strain with forebrain abnormalities (callosal defects); Phenome Project A list
5. C3H/HeJ
       Sequenced by Perlegen/NIEHS; paternal parent of the BXH panel; Phenome Project A list
6. C57BL/6J
       Sequenced by NHGRI; parental strain of AXB/BXA, BXD, and BXH; Phenome Project A list
7. C57BL/6ByJ
       Paternal substrain of B6 used to generate the CXB panel
8. CAST/Ei
       Collaborative Cross strain sequenced by NIEHS; Phenome Project A list
9. DBA/2J
       Sequenced by Perlegen/NIEHS and Celera; paternal parent of the BXD panel; Phenome Project A list
10. KK/HIJ
        Sequenced by Perlegen/NIEHS
11. LG/J
        Paternal parent of the LGXSM panel
12. NOD/LtJ
        Collaborative Cross strain sequenced by NIEHS; Phenome Project B list; diabetic
13. NZO/HILtJ
        Collaborative Cross strain
14. PWD/PhJ
        Sequenced by Perlegen/NIEHS; parental strain for a consomic set by Forjet and colleagues
15. PWK/PhJ
        Collaborative Cross strain; Phenome Project D list
16. WSB/EiJ
        Collaborative Cross strain sequenced by NIEHS; Phenome Project C list
17. B6D2F1 and D2B6F1
    F1 hybrids generated by crossing C57BL/6J with DBA/2J

We have not combined data from reciprocal F1s because they have different Y chromosome and mitochondial haplotypes. Parent-of-origin effects (imprinting, maternal environment) may also lead to interesting differences in hippocampal transcript levels.

These strains are available from The Jackson Laboratory. BXD43 through BXD100 are available from Lu Lu and colleagues at UTHSC.

    About the animals and tissue used to generate this set of data:

BXD animals were obtained from UTHSC, UAB, or directly from The Jackson Laboratory (see Table 1 below). Animals were housed at UTHSC, Beth Israel Deaconess, or the Jackson Laboratory before sacrifice. Virtually all CXB animals were obtained directly at the Jackson Laboratory by Lu Lu. We thanks Muriel Davission for making it possible to collect these cases on site. Standard inbred strain stock was from The Jackson Laboratory, but most animals were housed or reared at UTHSC. Mice were killed by cervical dislocation and brains were removed and placed in RNAlater prior to dissection. Cerebella and olfactory bulbs were removed; brains were hemisected, and both hippocampi were dissected whole by Hong Tao Zhang in the Lu lab. Hippocampal samples are very close to complete (see Lu et al., 2001) but probably include variable amounts of subiculum and fimbria.

A pool of dissected tissue typically from six hippocampi and three naive adults of the same strain, sex, and age was collected in one session and used to generate cRNA samples. Two-hundred and one RNA samples were extracted at UTHSC by Zhiping Jia, four samples by Shuhua Qi (R2331H1, R2332H1, P2350H1, R2349H1), and one by Siming Shou (R0129H2).

A great majority of animals used in this study were between 45 and 90 days of age (average of 66 days, maximum range from 41 to 196 days; see Table 1 below).

Sample Processing: Samples were processed in the INIA Bioanalytical Core at the W. Harry Feinstone Center for Genomic Research, The University of Memphis, led by Thomas R. Sutter. All processing steps were performed by Dr. Shirlean Goodwin. In brief, RNA purity was evaluated using the 260/280 nm absorbance ratio, and values had to be greater than 1.8. The majority of samples were 1.9 to 2.1. RNA integrity was assessed using the Agilent Bioanalyzer 2100. We required an RNA integrity number (RIN) of greater than 8. This RIN value is based on the intensity ratio and amplitude of 18S and 28S rRNA signals. The standard Eberwine T7 polymerase method was used to catalyze the synthesis of cDNA template from polyA-tailed RNA using Superscript II reverse transcriptase (Invitrogen Inc.). The Enzo Life Sciences, Inc., BioArray High Yield RNA Transcript Labeling Kit (T7, Part No. 42655) was used to synthesize labeled cRNA. The cRNA was evaluated using both the 260/280 ratio (values of 2.0 or 2.1 are acceptable) and the Bioanalyzer output (a dark cRNA smear on the 2100 output centered roughly between 600 and 2000 nucleotides is required). Those samples that passed both QC steps (10% usually failed and new RNA samples had to be acquired and processed) were then sheared using a fragmentation buffer included in the Affymetrix GeneChip Sample Cleanup Module (Part No. 900371). Fragmented cRNA samples were either stored at -80 deg. C until use or were immediately injected onto the array. The arrays were hybridized and washed following standard Affymetrix protocols.

Replication and Sample Balance: We obtained a male sample pool and female sample pool from each isogenic group. While all strains were orginally represented by matched male and female samples, not all data sets passed the final quality control steps. Seventy-seven of 99 strains are represented by pairs or (rarely) trios of arrays. The first and last samples are technical replicates of a B6D2F1 hippocampal pool (aliquotes R1291H3 and R1291H4).

Experimental Design and Batch Structure: This data set consists arrays processed in six groups over a three month period (May 2005 to August 2005). Each group consists of 32 to 34 arrays.Sex, strain, and strain type (BXD, CXB, and MDP) were interleaved among groups to ensure reasonable balance and to minimize group-by-strain statistical confounds in group normalization. The two independent samples from a single strain were always run in different groups. All arrays were processed using a single protocol by a single operator, Shirlean Goodwin.

All samples in a group were labeled on one day, except for a few cases that failed QC on their first pass. The hybridization station accommodates up to 20 samples, and for this reason each group was split into a large first set of 20 samples and a second set of 12 to 14 samples. Samples were washed in groups of four and then held in at 4 deg C until all 20 (or 12-14) arrays were ready to scan. The last four samples out of the wash stations were scanned directly. Samples were scanned in sets of four.

    Data Table 1:

This table lists all arrays by sample ID (tube ID), strain, age, sex, the name of the key Affymetrix CEL file, processing batch ID for the sample, whether or not the sample was stored in RNAlater prior to RNA extraction, number of animals in each sample pool (should read 3, not 1), the fraction of probe sets that generated values >2 standard deviation units from the mean (RMA 2Z outlier), and seven Affymetrix quality control values (scale factor, background, percent present, absent, and marginal, and the actin and Gadph 3' to 5' ratios. Finally, source provides information on the original source of tissues (Glenn = tissue dissected by Glenn D. Rosen, Beth Israel Deaconess Medical Center using mice received directly from JAX).

index tube ID strain age sex batch Id pool size RMA outlier scale factor back ground average present absent marginal AFFX-b-Actin AFFX-Gapdh source 1 R2028H2 129S1/SvImJ 66 F 5 3 0.1 4.362 64.49 0.497 0.484 0.019 2.78 1.13 JAX 2 R2029H2 129S1/SvImJ 66 M 6 3 0.04 5.208 41.21 0.49 0.49 0.02 1.62 0.95 JAX 3 R2030H1 A/J 57 M 5 2 0.06 3.307 45.16 0.527 0.454 0.018 1.63 0.99 UTM RW 4 R2032H3 AKR/J 66 F 5 3 0.04 3.054 61.03 0.51 0.471 0.018 1.46 0.79 JAX 5 R2454H1 AKR/J 66 M 6 4 0.11 2.892 58.55 0.474 0.507 0.019 1.99 0.78 JAX 6 R1289H2 B6D2F1 64 F 6 3 0.02 2.406 53.84 0.492 0.489 0.019 1.61 0.96 UTM RW 7 R1291H3 B6D2F1 66 M 1 3 0.01 3.524 48.54 0.487 0.494 0.019 1.21 1.52 UTM RW 8 R1291H4 B6D2F1 66 M 6 3 0.08 3.891 46.69 0.512 0.469 0.019 1.9 0.89 UTM RW 9 R2036H3 BALB/cJ 51 F 5 3 0.12 2.611 56.29 0.518 0.466 0.017 3.3 1.23 UTM RW 10 R2053H1 BALB/cJ 55 M 5 3 0.1 2.505 63.27 0.499 0.483 0.018 3.1 1.34 UTM RW 11 R2037H2 BALB/cJ 51 M 6 4 0.01 2.546 58.13 0.497 0.485 0.018 1.26 0.77 UTM RW 12 R1507H1 BXD1 58 M 3 3 0.02 4.056 60.17 0.478 0.503 0.019 1.15 0.76 Glenn 13 R1520H1 BXD2 56 F 4 4 0.09 1.715 71.62 0.515 0.467 0.018 2.36 1.6 Glenn 14 R1516H1 BXD2 61 M 1 4 0.01 2.231 64.86 0.508 0.474 0.019 1.3 1.53 Glenn 15 R1593H2 BXD5 60 F 1 4 0 1.913 59.96 0.487 0.493 0.02 0.98 1.44 Glenn 16 R1692H1 BXD5 60 M 3 2 0.07 3.764 72.74 0.465 0.516 0.02 1.15 0.74 Glenn 17 R1539H2 BXD6 59 F 1 4 0 2.488 54.97 0.518 0.463 0.018 1.08 1.33 Glenn 18 R1538H1 BXD6 59 M 4 3 0.01 2.585 50.27 0.505 0.475 0.02 1.46 0.79 Glenn 19 R1518H1 BXD8 56 F 1 3 0 2.92 54.84 0.515 0.465 0.02 1.32 1.24 Glenn 20 R1548H1 BXD8 59 M 6 3 0.07 2.132 59.37 0.504 0.477 0.019 2.16 1.54 Glenn 21 R1350H2 BXD9 86 F 1 3 0.05 2.771 60.62 0.5 0.482 0.018 1.01 1.28 UMemphis 22 R1531H1 BXD11 56 F 6 3 0.06 2.229 56.36 0.505 0.475 0.02 2.23 1.02 Glenn 23 R1367H1 BXD11 56 M 1 3 0.01 2.11 78.78 0.503 0.477 0.02 1.07 1.27 Glenn 24 R1530H1 BXD12 58 F 1 3 0 3.227 53.77 0.505 0.477 0.018 0.95 1.4 Glenn 25 R1529H1 BXD13 58 F 6 3 0.05 2.55 59.05 0.497 0.485 0.018 2 1.54 Glenn 26 R1662H2 BXD13 60 M 1 3 0.03 4.603 45.81 0.509 0.472 0.019 1.3 0.82 Glenn 27 R1524H1 BXD15 60 F 6 4 0.02 2.961 50.93 0.497 0.484 0.019 1.74 0.91 Glenn 28 R1515H1 BXD15 61 M 1 3 0.01 3.316 57.05 0.503 0.478 0.019 1.32 1.21 Glenn 29 R1661H1 BXD16 61 F 1 3 0.01 2.778 59.81 0.516 0.466 0.019 1.39 1.2 Glenn 30 R1594H1 BXD16 61 M 4 3 0.03 2.634 53.66 0.504 0.478 0.018 1.96 1.51 Glenn 31 R1471H1 BXD19 157 M 1 3 0.02 3.165 43.34 0.519 0.462 0.018 1.01 1.29 UTM JB 32 R1573H1 BXD20 59 F 1 3 0.02 3.749 52.7 0.513 0.469 0.018 1.01 1.27 Glenn 33 R2507H1 BXD20 60 M 6 3 0.06 3.568 57 0.472 0.508 0.02 1.29 0.76 Glenn 34 R1347H2 BXD21 64 F 1 4 0.01 2.881 61.49 0.494 0.486 0.019 0.92 1.22 UMemphis 35 R1337H2 BXD21 102 F 2 4 0 2.673 58.05 0.492 0.489 0.019 1.4 0.76 UAB 36 R1848H3 BXD22 196 F 6 4 0.02 2.943 51.7 0.494 0.485 0.021 2.2 0.78 UAB 37 R1525H1 BXD22 59 M 2 3 0.02 2.248 55.76 0.548 0.433 0.018 1.26 0.74 Glenn 38 R1280H2 BXD23 56 F 1 3 0.01 3.187 54.63 0.458 0.523 0.019 0.96 1.2 UTM RW 39 R1537H1 BXD23 58 F 5 3 0.1 3.719 67.54 0.468 0.513 0.019 1.51 0.96 Glenn 40 R1343H2 BXD24 71 F 2 3 0.01 2.083 65.07 0.506 0.474 0.019 1.46 0.75 UMemphis 41 R1517H1 BXD24 57 M 3 3 0.01 3.471 53.66 0.504 0.476 0.019 1.28 0.78 Glenn 42 R1366H1 BXD27 60 F 2 4 0 2.26 48.46 0.518 0.463 0.019 1.29 0.77 Glenn 43 R1849H1 BXD27 70 M 5 3 0.06 8.801 38.34 0.468 0.512 0.019 2.42 1.08 UAB 44 R1353H1 BXD28 79 F 3 4 0.01 3.22 76.22 0.48 0.5 0.02 1.33 0.78 UMemphis 45 R2332H1 BXD28 60 M 2 3 0.01 3.217 63.68 0.491 0.49 0.019 1.37 0.79 Glenn 46 R1532H1 BXD29 57 F 2 3 0.01 2.122 59.18 0.524 0.456 0.019 1.17 0.76 Glenn 47 R1356H1 BXD29 76 M 5 3 0.01 4.033 47.67 0.52 0.463 0.017 1.17 0.78 UMemphis 48 R1240H2 BXD31 61 M 2 3 0.02 2.335 65.17 0.507 0.474 0.019 1.31 0.78 UTM RW 49 R1508H2 BXD32 58 M 2 4 0.01 1.917 67.78 0.539 0.442 0.019 1.28 0.73 Glenn 50 R1345H3 BXD33 65 F 2 2 0.01 2.098 63.14 0.522 0.459 0.019 1.27 0.73 UMemphis 51 R1581H1 BXD33 59 M 3 3 0.01 3.229 53.16 0.496 0.485 0.019 1.19 0.78 Glenn 52 R1527H1 BXD34 59 F 2 3 0.01 2.3 58.92 0.51 0.471 0.019 1.24 0.76 Glenn 53 R1339H3 BXD34 74 M 5 3 0.12 2.888 53.49 0.506 0.476 0.018 2.39 1.35 UMemphis 54 R1469H1 BXD36 83 F 3 3 0.02 3.473 49.9 0.494 0.486 0.02 1.11 0.76 UMemphis 55 R1363H1 BXD36 77 M 2 4 0.01 2.184 48.19 0.538 0.443 0.02 1.28 0.77 UMemphis 56 R1855H1 BXD38 55 F 3 4 0.01 3.536 54.54 0.49 0.492 0.018 1.39 0.75 Glenn 57 R1510H1 BXD38 59 M 2 3 0.01 2.186 68.06 0.521 0.46 0.019 1.26 0.79 Glenn 58 R1528H2 BXD39 59 F 2 3 0.03 4.717 38.3 0.511 0.47 0.02 1.12 0.75 Glenn 59 R1514H1 BXD39 59 M 3 3 0.03 3.992 56.06 0.477 0.504 0.019 1.43 0.81 Glenn 60 R1522H1 BXD40 59 F 4 4 0 2.631 67.16 0.49 0.491 0.018 1.56 0.77 Glenn 61 R1359H1 BXD40 73 M 2 3 0.09 7.458 39.86 0.451 0.527 0.021 1.28 0.74 UMemphis 62 R1334H2 BXD43 59 F 1 3 0 2.672 54.36 0.492 0.491 0.017 1.2 2.06 UTM RW 63 R1303H1 BXD43 63 M 3 4 0.02 3.497 51.9 0.486 0.495 0.019 1.15 0.8 UTM RW 64 R1326H1 BXD44 65 F 3 4 0 3.412 53.96 0.496 0.485 0.018 1.35 0.78 UTM RW 65 R1577H2 BXD44 56 M 1 3 0.02 2.159 67.52 0.512 0.469 0.019 1.18 1.71 UTM RW 66 R1316H1 BXD48 58 F 4 3 0 2.445 68.59 0.515 0.467 0.019 1.16 0.73 UTM RW 67 R1575H3 BXD48 65 M 3 4 0.05 4.577 55.78 0.466 0.514 0.019 1.59 0.9 UTM RW 68 R2521H1 BXD50 63 F 6 4 0.01 3.109 57.28 0.495 0.485 0.02 1.23 0.78 UTM RW 69 R1944H2 BXD50 81 M 1 3 0.01 2.546 63.39 0.495 0.485 0.02 0.9 1.57 UTM RW 70 R2331H1 BXD51 66 F 3 3 0.03 3.534 44.42 0.501 0.481 0.017 1.2 0.9 UTM RW 71 R1582H1 BXD51 71 M 6 4 0.03 2.92 47.87 0.489 0.491 0.02 1.36 0.75 UTM RW 72 R1331H1 BXD60 60 F 4 3 0.01 2.867 50.33 0.492 0.487 0.021 1.34 0.78 UTM RW 73 R1281H2 BXD60 59 M 1 3 0 2.39 58.44 0.511 0.469 0.02 0.94 1.2 UTM RW 74 R1856H2 BXD61 94 M 1 2 0 3.502 49.6 0.501 0.48 0.019 0.96 1.3 UTM RW 75 R1246H1 BXD62 54 F 1 4 0.02 3.405 51.47 0.511 0.471 0.018 1.14 1.34 UTM RW 76 R1585H2 BXD62 64 M 6 4 0.01 3.156 55.77 0.518 0.464 0.018 1.43 0.82 UTM RW 77 R1945H1 BXD63 107 F 1 3 0.02 2.811 52.65 0.522 0.459 0.019 1.05 1.36 UTM RW 78 R2093H3 BXD63 70 M 6 3 0.02 3.894 42.85 0.503 0.477 0.019 1.29 1.01 UTM RW 79 R2062H2 BXD64 65 F 1 3 0.05 3.795 78.48 0.513 0.468 0.019 0.98 1.43 UTM RW 80 R2061H1 BXD64 87 M 3 4 0.01 3.536 61.57 0.477 0.504 0.019 1.31 0.78 UTM RW 81 R2054H2 BXD65 55 F 1 2 0.03 3.159 80.96 0.48 0.502 0.018 1.09 1.24 UTM RW 82 R2056H2 BXD65 89 M 6 2 0 2.836 59.6 0.504 0.477 0.019 1.3 0.75 UTM RW 83 R1941H2 BXD66 78 F 1 4 0.01 2.734 50.93 0.499 0.481 0.02 1.18 1.29 UTM RW 84 R1949H2 BXD66 96 M 4 2 0.04 2.828 51.27 0.474 0.508 0.019 2.05 1.12 UTM RW 85 R2060H1 BXD67 54 F 6 3 0.01 2.561 43.88 0.502 0.479 0.02 1.7 0.84 UTM RW 86 R2052H1 BXD67 61 M 1 4 0.01 3.161 43.23 0.521 0.46 0.018 1.09 1.31 UTM RW 87 R2074H1 BXD68 60 F 5 3 0.02 6.528 49.62 0.479 0.502 0.019 1.48 0.83 UTM RW 88 R1928H1 BXD68 72 M 2 2 0.01 2.404 48.28 0.521 0.459 0.02 1.3 0.74 UTM RW 89 R1439H3 BXD69 60 F 2 3 0.02 2.463 59.14 0.522 0.459 0.018 1.31 0.78 UTM RW 90 R1559H1 BXD69 64 M 3 3 0.03 2.987 67.74 0.486 0.496 0.017 1.38 0.8 UTM RW 91 R2134H1 BXD70 64 F 5 2 0.02 2.148 58.64 0.532 0.45 0.019 1.4 0.85 UTM RW 92 R2063H1 BXD70 55 M 2 3 0.02 3.481 55.32 0.513 0.469 0.018 1.28 0.71 UTM RW 93 R1277H1 BXD73 60 F 4 2 0.01 2.576 62.45 0.502 0.479 0.019 1.35 0.79 UTM RW 94 R1443H2 BXD73 76 M 2 3 0.01 2.312 64.34 0.499 0.481 0.02 1.48 0.77 UTM RW 95 R2055H2 BXD74 79 M 2 3 0.01 2.576 56.84 0.509 0.473 0.018 1.46 0.88 UTM RW 96 R2316H1 BXD74 193 M 5 2 0.01 3.457 55.35 0.508 0.471 0.02 1.17 0.78 UTM RW 97 R1871H1 BXD75 61 F 2 3 0.04 1.723 56.4 0.53 0.451 0.019 1.3 0.76 UTM RW 98 R1844H2 BXD75 90 M 3 4 0.01 1.934 56.23 0.52 0.461 0.019 1.62 0.86 UTM RW 99 R1948H2 BXD76 81 F 2 3 0.01 1.507 68.85 0.553 0.428 0.02 1.3 0.75 UTM RW 100 R2094H1 BXD76 61 M 5 4 0.01 3.299 42.69 0.519 0.462 0.019 1.39 0.88 UTM RW 101 R2262H1 BXD77 62 F 3 4 0.02 4.317 47.16 0.493 0.488 0.019 1.32 0.74 UTM RW 102 R1423H1 BXD77 62 M 2 3 0.02 3.071 54.15 0.51 0.471 0.019 1.26 0.74 UTM RW 103 R1947H1 BXD79 108 F 2 2 0.01 2.599 51.52 0.524 0.457 0.019 1.35 0.74 UTM RW 104 R2092H1 BXD79 86 M 5 4 0.06 3.735 42.25 0.514 0.468 0.018 2.94 1.06 UTM RW 105 R1880H1 BXD80 68 F 5 3 0.06 4.855 42.22 0.501 0.481 0.018 2.17 1.36 UTM RW 106 R1881H2 BXD80 68 M 2 3 0.02 2.073 48.93 0.524 0.458 0.019 1.34 0.83 UTM RW 107 R2075H1 BXD83 60 F 2 3 0.01 2.454 55.1 0.502 0.48 0.018 1.27 0.77 UTM RW 108 R2076H2 BXD83 60 M 6 3 0.03 2.624 55.65 0.495 0.488 0.018 2.21 0.94 UTM RW 109 R2077H2 BXD84 62 F 6 2 0 2.1 71.87 0.522 0.459 0.018 1.68 0.81 UTM RW 110 R2135H3 BXD84 75 M 2 2 0.01 2.467 64.46 0.505 0.476 0.019 1.2 0.74 UTM RW 111 R1473H1 BXD85 79 F 2 3 0.02 3.384 55.34 0.478 0.502 0.02 1.24 0.77 UTM RW 112 R1474H1 BXD85 57 M 1 3 0.01 2.831 55.24 0.522 0.461 0.018 1.04 1.29 UTM RW 113 R1597H1 BXD85 86 M 4 4 0.09 2.028 53.95 0.487 0.492 0.021 1.28 0.83 UTM RW 114 R1415H1 BXD86 77 F 4 3 0.02 2.525 53.16 0.495 0.485 0.02 1.66 0.91 UTM RW 115 R1710H1 BXD87 84 M 2 4 0.01 2.697 56.4 0.512 0.469 0.019 1.28 0.79 UTM RW 116 R1872H2 BXD89 90 F 2 2 0.02 3.013 63.53 0.492 0.488 0.021 1.22 0.72 UTM RW 117 R1850H3 BXD89 82 M 4 4 0.03 2.736 44.89 0.498 0.483 0.019 1.5 0.83 UTM RW 118 R2058H1 BXD90 61 F 2 3 0.01 3.389 48.05 0.502 0.478 0.02 1.53 0.76 UTM RW 119 R1301H2 BXD92 58 F 2 3 0.02 3.543 41.97 0.522 0.46 0.018 1.5 0.79 UTM RW 120 R1309H1 BXD92 59 M 4 3 0.05 1.655 66.34 0.498 0.481 0.021 1.52 0.82 UTM RW 121 R2057H1 BXD93 92 F 5 3 0.02 4.033 44.41 0.509 0.471 0.02 1.22 0.78 UTM RW 122 R2059H1 BXD93 58 M 1 3 0 3.058 60.29 0.493 0.488 0.019 1.18 1.37 UTM RW 123 R2313H1 BXD94 59 F 3 3 0 3.091 59.45 0.487 0.495 0.018 1.34 0.73 UTM RW 124 R1915H1 BXD96 65 F 5 2 0.04 5.145 46.19 0.502 0.481 0.017 1.37 0.74 UTM RW 125 R1846H2 BXD96 63 M 1 3 0 3.159 55.85 0.487 0.493 0.02 0.92 1.26 UTM RW 126 R1927H2 BXD97 67 M 1 3 0.04 2.622 57.81 0.539 0.444 0.017 1.45 1.32 UTM RW 127 R1942H1 BXD98 62 F 5 3 0.04 3.104 48.42 0.528 0.454 0.019 2.22 1.08 UTM RW 128 R1943H2 BXD98 62 M 3 3 0.02 4.04 56.85 0.484 0.497 0.019 1.18 0.76 UTM RW 129 R2197H1 BXD99 70 F 3 3 0.02 4.288 51.75 0.49 0.492 0.018 1.35 0.81 UTM RW 130 R2315H1 BXD99 84 M 5 2 0.03 6.036 43.05 0.484 0.497 0.018 1.7 0.96 UTM RW 131 R2038H3 C3H/HeJ 63 F 6 3 0.02 2.671 66.74 0.476 0.504 0.02 1.41 0.77 UTM RW 132 R2039H1 C3H/HeJ 63 M 5 3 0.1 3.384 44.15 0.528 0.454 0.017 2.16 0.88 UTM RW 133 R2137H1 C57BL/6ByJ 55 F 5 3 0.02 4.746 47.01 0.488 0.493 0.018 1.23 0.79 JAX 134 R1361H1 C57BL/6J 69 F 6 4 0.01 3.058 51.87 0.477 0.503 0.02 1.67 0.76 UTM RW 135 R2041H2 C57BL/6J 65 M 1 4 0.04 3.341 49.26 0.527 0.456 0.018 1.14 1.45 UTM RW 136 R1449H2 C57BL/6J 71 M 5 3 0.09 3.592 44.32 0.47 0.51 0.02 1.68 0.77 UTM DG 137 R2619H1 CAST/Ei 64 F 5 3 0.14 4.077 51.87 0.455 0.528 0.018 2.74 1.2 JAX 138 R2116H1 CXB1 55 F 3 3 0.07 5.792 51.59 0.459 0.521 0.02 1.17 0.8 JAX 139 R2096H1 CXB1 55 M 4 2 0.01 3.435 53.78 0.495 0.485 0.02 1.22 0.79 JAX 140 R2124H1 CXB10 53 F 4 2 0.11 4.867 39.88 0.451 0.528 0.02 1.55 0.8 JAX 141 R2125H1 CXB11 58 F 3 3 0.03 3.256 54.95 0.461 0.519 0.02 1.46 0.77 JAX 142 R2128H1 CXB11 58 M 4 2 0.06 4.986 54.13 0.465 0.515 0.02 1.11 0.83 JAX 143 R2126H1 CXB12 47 F 4 3 0.11 3.935 54.11 0.469 0.511 0.021 1.5 0.79 JAX 144 R2109H1 CXB12 47 M 3 3 0.07 4.518 49.26 0.488 0.492 0.02 1.23 0.77 JAX 145 R2110H1 CXB13 56 M 4 3 0.21 3.478 48.08 0.461 0.517 0.022 1.21 0.78 JAX 146 R2117H2 CXB2 62 F 4 2 0.04 3.39 45.97 0.533 0.45 0.017 2.05 0.89 JAX 147 R2098H1 CXB2 68 M 3 3 0.02 2.572 54.22 0.496 0.485 0.019 1.38 0.86 JAX 148 R2118H1 CXB3 47 F 3 3 0.03 3.646 63.16 0.478 0.503 0.019 1.22 0.77 JAX 149 R2100H1 CXB3 47 M 4 3 0.02 5.76 51.38 0.48 0.503 0.017 1.24 0.81 JAX 150 R2119H1 CXB4 58 F 4 3 0.02 3.897 49.21 0.488 0.494 0.018 1.31 0.79 JAX 151 R2101H1 CXB4 58 M 3 3 0.13 7.372 53.77 0.433 0.548 0.019 1.2 0.97 JAX 152 R2505H1 CXB5 80 F 6 3 0.02 2.83 49.6 0.499 0.48 0.02 1.33 0.76 UTM RW 153 R2131H1 CXB5 42 M 4 3 0.1 5.577 51.15 0.434 0.547 0.019 1.7 0.89 JAX 154 R0129H2 CXB5 70 M 3 3 0.07 4.829 45.42 0.488 0.493 0.019 1.23 0.83 UTM RW 155 R2102H1 CXB6 49 M 4 3 0.07 5.148 51.63 0.453 0.529 0.018 1.43 0.87 JAX 156 R2121H1 CXB7 63 F 4 2 0.06 4.904 48.71 0.464 0.517 0.019 1.19 0.92 JAX 157 R2104H2 CXB7 58 M 3 2 0.06 3.389 48.79 0.502 0.479 0.019 1.74 1.48 JAX 158 R2122H1 CXB8 54 F 3 3 0.04 4.128 59.77 0.451 0.529 0.02 1.12 0.76 JAX 159 R2105H1 CXB8 41 M 4 3 0.16 3.146 61.04 0.451 0.53 0.019 1.34 0.84 JAX 160 R2123H1 CXB9 54 F 3 3 0.08 5.708 55.94 0.438 0.543 0.019 1.32 0.78 JAX 161 R2106H1 CXB9 54 M 4 3 0.06 5.868 46.55 0.469 0.512 0.019 1.18 0.82 JAX 162 R2045H2 D2B6F1 65 F 1 2 0.01 4.403 47.99 0.497 0.485 0.018 1.09 1.53 UTM RW 163 R1595H2 D2B6F1 63 F 5 3 0.06 2.579 58.49 0.506 0.475 0.019 2.49 1.21 UTM RW 164 R1551H1 D2B6F1 72 F 6 3 0.02 2.62 53.76 0.506 0.476 0.018 1.37 0.76 UTM RW 165 R1468H1 DBA/2J 64 F 5 3 0.03 2.929 53.8 0.515 0.465 0.019 1.28 0.79 UTM RW 166 R1683H1 KK/HIJ 72 F 6 3 0.02 3.919 54.23 0.491 0.489 0.02 1.31 0.83 JAX 167 R1687H3 KK/HIJ 72 M 5 3 0.04 3.888 40.86 0.499 0.483 0.019 1.86 0.88 JAX 168 R2046H1 LG/J 63 F 5 2 0.03 2.822 59.18 0.514 0.468 0.018 1.68 0.8 UTM RW 169 R2047H2 LG/J 63 M 6 3 0.07 2.038 60.34 0.509 0.471 0.02 2.16 0.95 UTM RW 170 R2048H1 NOD/LtJ 77 F 6 2 0.14 4.045 50.21 0.489 0.49 0.021 2.89 0.95 UTM RW 171 R2049H3 NOD/LtJ 76 M 5 3 0.1 2.328 52.78 0.519 0.462 0.019 3.09 1.35 UTM RW 172 R2200H1 NZO/HlLtJ 62 F 5 2 0.03 2.648 54.29 0.543 0.438 0.019 1.27 0.8 JAX 173 R2350H1 NZO/HlLtJ 96 M 6 2 0.19 2.391 50.52 0.518 0.463 0.02 3.71 2.21 JAX 174 R2051H3 PWD/PhJ 64 M 5 3 0.07 3.266 51.5 0.475 0.506 0.019 2.8 1.01 UTM RW 175 R2322H1 PWK/PhJ 63 F 5 2 0.09 2.94 54.91 0.511 0.47 0.019 2.32 1.02 JAX 176 R2349H1 PWK/PhJ 83 M 6 2 0.15 3.306 54.93 0.459 0.522 0.019 4.65 1.45 JAX 177 R2198H2 WSB/EiJ 58 F 6 1 0.02 2.922 57.97 0.502 0.479 0.019 1.44 0.76 JAX 178 R2199H1 WSB/EiJ 58 M 5 3 0.04 3.171 54.95 0.475 0.505 0.02 1.32 0.81 JAX

    Downloading all data:

All data tables will be made active as sooon as the global analysis by the Hippocampus Consoritum has been accepted for publication. Please see text on Data Sharing Policies, and Conditions and Limitations, and Contacts. Following publication, download a summary text file or Excel file of the PDNN probe set data. Contact RW Williams regarding data access problems.

    About the array platform:

Affymetrix Mouse Genome 430 2.0 array: The 430v2 array consists of 992936 useful 25-nucleotide probes that estimate the expression of approximately 39,000 transcripts and the majority of known genes and expressed sequence tags. The array sequences were selected late in 2002 using Unigene Build 107 by Affymetrix. The UTHSC group has recently reannotated all probe sets on this array, producing more accurate data on probe and probe set targets. All probes were aligned to the most recent assembly of the Mouse Genome (Build 34, mm6) using Jim Kent's BLAT program. Many of the probe sets have been manually curated by Jing Gu and Rob Williams.

    About data processing:

Harshlight was used to examine the image quality of the array (CEL files). Bad areas (bubbles, scratches, blemishes) of arrays were masked.

First pass data quality control: Affymetrix GCOS provides useful array quality control data including:

1. The scale factor used to normalize mean probe intensity. This averaged 3.3 for the 179 arrays that passed and 6.2 for arrays that were excluded. The scale factor is not a particular critical parameter.
2. The average background level. Values averaged 54.8 units for the data sets that passed and 55.8 for data sets that were excluded. This factor is not important for quality control.
3. The percentage of probe sets that are associated with good signal ("present" calls). This averaged 50% for the 179 data sets that passed and 42% for those that failed. Values for passing data sets extended from 43% to 55%. This is a particularly important criterion.
4. The 3':5' signal ratios of actin and Gapdh. Values for passing data sets averaged 1.5 for actin and 1.0 for Gapdh. Values for excluded data sets averaged 12.9 for actin and 9.6 for Gapdh. This is a highly discriminative QC criterion, although one must keep in mind that only two transcripts are being tested. Sequence variation among strains (particularly wild derivative strains such as CAST/Ei) may affect these ratios.

The second step in our post-processing QC involves a count of the number of probe sets in each array that are more than 2 standard deviations (z score units) from the mean across the entire 206 array data sets. This was the most important criterion used to eliminate "bad" data sets. All 206 arrays were processed togther using standard RMA and PDNN methods. The count and percentage of probe sets in each array that were beyond the 2 z theshold was computed. Using the RMA transform the average percentage of probe sets beyond the 2 z threshold for the 179 arrays that finally passed of QC procedure was 1.76% (median of 1.18%). In contrast the 2 z percentage was more than 10-fold higher (mean of 22.4% and median 20.2%) for those arrays that were excluded. This method is not very senstive to the transformation method that is used. Using the PDNN transform the average percent of probe sets exceeding was 1.31% for good arrays and was 22.6% for those that were excluded. In our opinion, this 2 z criterion is the most useful criterion for the final decision of whether or not to include arrays, although again, allowances need to be made for wild strains that one expects to be different from the majority of conventional inbred strains. For examploe, if a data set has excellent characteristics on all of the Affymetrix GCOS metrics listed above, but generates a high 2 z percentage, then one whould include the ssample if one can verify that there are no problems in sample and data set identification.

The entire procedure can be reapplied once the initial outlier data sets have been eliminated to detect any remaining outlier data sets.

DataDesk was used to examine the statistical quality of the of the probe level (CEL) data after step 5 below. DataDesk allows a rapid detection of subsets of probes that are particular sensitive to still unknown factors in array processing. Arrays can then be categorized at the probe level into "reaction classes." A reaction class is a group of arrays for which the expression of essentially all probes are colinear over the full range of log2 values. A single but large group of arrays (n = 32) processed in essentially the identical manner by a single operator can produce arrays belonging to as many as four different reaction classes. Reaction classes are NOT related to strain, age, sex, treatment, or any known biological parameter (technical replicates can belong to different reaction classes). We do not yet understand the technical origins of reaction classes. The number of probes that contribute to the definition of reaction classes is quite small (<10% of all probes). We have categorized all arrays in this data set into one of 5 reaction classes. These have then been treated as if they were separate batches. Probes in these data type "batches" have been aligned to a common mean as described below.

Probe (cell) level data from the CEL file: These CEL values produced by GCOS are 75% quantiles from a set of 91 pixel values per cell. <0L>

We added an offset of 1.0 unit to each cell signal to ensure that all values could be logged without generating negative values. We then computed the log base 2 of each cell.

We performed a quantile normalization of the log base 2 values for all arrays using the same initial steps used by the RMA transform.

We computed the Z scores for each cell value.

We multiplied all Z scores by 2.

We added 8 to the value of all Z scores. The consequence of this simple set of transformations is to produce a set of Z scores that have a mean of 8, a variance of 4, and a standard deviation of 2. The advantage of this modified Z score is that a two-fold difference in expression level (probe brightness level) corresponds approximately to a 1 unit difference.

inally, we computed the arithmetic mean of the values for the set of microarrays for each strain. Technical replicates were averaged before computing the mean for independent biological samples. Note, that we have not (yet) corrected for variance introduced by differences in sex or any interaction terms. We have not corrected for background beyond the background correction implemented by Affymetrix in generating the CEL file. We eventually hope to add statistical controls and adjustments for some of these variables.

Probe set data from the CHP file: The expression values were generated using PDNN. The same simple steps described above were also applied to these values. Every microarray data set therefore has a mean expression of 8 with a standard deviation of 2. A 1 unit difference represents roughly a two-fold difference in expression level. Expression levels below 5 are usually close to background noise levels.

Probe level QC: Log2 probe data of all arrays were inspected in DataDesk before and after quantile normalization. Inspection involved examining scatterplots of pairs of arrays for signal homogeneity (i.e., high correlation and linearity of the bivariate plots) and looking at all pairs of correlation coefficients. XY plots of probe expression and signal variance were also examined. Probe level array data sets were organized into reaction groups. Arrays with probe data that were not homogeneous when compared to other arrays were flagged.

Probe set level QC: The final normalized individual array data were evaluated for outliers. This involved counting the number of times that the probe set value for a particular array was beyond two standard deviations of the mean. This outlier analysis was carried out using the PDNN, RMA and MAS5 transforms and outliers across different levels of expression. Arrays that were associated with an average of more than 8% outlier probe sets across all transforms and at all expression levels were eliminated. In contrast, most other arrays generated fewer than 5% outliers.

Validation of strains and sex of each array data set: A subset of probes and probe sets with a Mendelian pattern of inheritance were used to construct a expression correlation matrix for all arrays and the ideal Mendelian expectation for each strain constructed from the genotypes. There should naturally be a very high correlation in the expression patterns of transcripts with Mendelian phenotypes within each strain, as well as with the genotype strain distribution pattern of markers for the strain.

Sex of the samples was validated using sex-specific probe sets such as Xist and Dby.

    Data source acknowledgment:

Data were generated with funds provided by a variety of public and private source to members of the Consortium. All of us thank Muriel Davisson, Cathy Lutz, and colleagues at the Jackson Laboratory for making it possible for us to add all of the CXB strains, and one or more samples from KK/HIJ, WSB/Ei, NZO/HILtJ, LG/J, CAST/Ei, PWD/PhJ, and PWK/PhJ to this study. We thank Yan Cui at UTHSC for allowing us to use his Linux cluster to align all M430 2.0 probes and probe sets to the mouse genome. We thank Hui-Chen Hsu and John Mountz for providing us BXD tissue samples, as well as many strains of BXD stock. We thanks Douglas Matthews (UMem in Table 1) and John Boughter (JBo in Table 1) for sharing BXD stock with us. Members of the Hippocampus Consortium thank the following sources for financial support of this effort:

• David C. Airey, Ph.D.
  Grant Support: Vanderbilt Institute for Integratie Genomics
  Department of Pharmacology
  david.airey at vanderbilt.edu
• Lu Lu, M.D.
  Grant Support: NIH U01AA13499, U24AA13513
• Fred H. Gage, Ph.D.
  Grant Support: NIH XXXX
• Dan Goldowitz, Ph.D.
  Grant Support: NIAAA INIA AA013503
  University of Tennessee Health Science Center
  Dept. Anatomy and Neurobiology
  email: dgold@nb.utmem.edu
• Shirlean Goodwin, Ph.D.
  Grant Support: NIAAA INIA U01AA013515
• Gerd Kempermann, M.D.
  Grant Support: The Volkswagen Foundation Grant on Permissive and Persistent Factors in Neurogenesis in the Adult Central Nervous System
  Humboldt-Universitat Berlin
  Universitatsklinikum Charite
  email: gerd.kempermann at mdc-berlin.de
• Kenneth F. Manly, Ph.D.
  Grant Support: NIH P20MH062009 and U01CA105417
• Richard S. Nowakowski, Ph.D.
  Grant Support: R01 NS049445-01
• Yanhua Qu, Ph.D.
  Grant Support: NIH U01CA105417
• Glenn D. Rosen, Ph.D.
  Grant Support: NIH P20
• Leonard C. Schalkwyk, Ph.D.
  Grant Support: MRC Career Establishment Grant G0000170
  Social, Genetic and Developmental Psychiatry
  Institute of Psychiatry,Kings College London
  PO82, De Crespigny Park London SE5 8AF
  L.Schalkwyk@iop.kcl.ac.uk
• Guus Smit, Ph.D.
  Dutch NeuroBsik Mouse Phenomics Consortium
  Center for Neurogenomics & Cognitive Research
  Vrije Universiteit Amsterdam, The Netherlands
  e-mail: guus.smit at falw.vu.nl
  Grant Support: BSIK 03053
• Thomas Sutter, Ph.D.
  Grant Support: INIA U01 AA13515 and the W. Harry Feinstone Center for Genome Research
• Stephen Whatley, Ph.D.
  Grant Support: XXXX
• Robert W. Williams, Ph.D.
  Grant Support: NIH U01AA013499, P20MH062009, U01AA013499, U01AA013513

    About this text file:

This text file originally generated prospectively by RWW on July 30 2005. Updated by RWW July 31, 2005.