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. 2023 Oct 19;11:1270892. doi: 10.3389/fcell.2023.1270892

TABLE 2.

Wide search for genes correlated to the proliferative activation of stem cells: selection of the 866 differentially expressed GLM genes whose expression was significantly changed in the dentate gyrus by running in both Set A and Set E (termed Set A_Set E_GLM). This analysis is parallel to, but wider than, the one shown for Set D_Set E_GLM in Table 1. In fact the 866 genes identified by the GLM procedure, i.e., whose expression was significantly changed by the interaction between running and genotype, were further selected for genes changed by running with p < 0.05 in both pairwise comparisons of Set A (p16 KO RUN vs p16 KO CTL) and of Set E (p16 KO RUN vs p16 WT RUN). The total number of genes identified in this way was 106: this group was named Set A_Set E_GLM selection and its differential gene regulation reproduces the pattern of proliferative activation of p16 KO stem cells by running, with respect to the other groups. Moreover, Set A_Set E_GLM selection includes all genes of Set D_Set E_GLM shown in Table 1, indicating that the Set A_Set E_GLM is more comprehensive. The list shown here is a selection among the Set A_Set E_GLM genes of those that could be functionally correlated, according to the existing literature, to the activation of stem and progenitor cells of the dentate gyrus, sorted by decreasing log2 fold change in Set A. The full list of 106 genes of Set D_Set E_GLM is shown in Supplementary Table S6. The genes highlighted in red are those with significant differential expression also in Set B (p16 KO CTL vs p16 WT CTL groups), i.e., the set representing the genetic signature of p16 knockout. p-values for Set A and Set E genes were obtained by DESeq2 analysis through the Wald test, while p-values for genes selected by the GLM model were obtained by testing the difference between the ‘full’ and ‘reduced’ model (Love et al., 2014).

Set A Set E GLM
KO RUN vs KO CTL KO RUN vs WT RUN
Gene name log2Fold change P-value log2Fold change P-value P-value Gene function correlated to stem cell activation References
Tfap2c 2.5099 0.0041173 2.1996 0.022623 0.0017784 Promotes hippocampal neurogenesis Mateus-Pinheiro et al. (2017)
Npw 1.8813 0.0005558 1.0357 0.043069 0.043133 Hippocampal signaling Chottova Dvorakova (2018)
Nlrc5 1.7969 0.0000344 0.91737 0.045703 0.0014689 Negative ROS regulator Li et al. (2021a)
Mki67 1.5041 0.0000001 1.487 2.12E-05 0.00055784 Cell cycle regulator Scholzen and Gerdes (2000)
Lepr 1.4658 0.0000007 0.80141 0.001078 0.0041875 Synaptic plasticity regulator Dhar et al. (2014)
Clcn1 1.3064 0.0079431 0.74279 0.0053492 0.0098998 Synaptic regulator Imbrici et al. (2015)
Cyp11a1 1.1998 0.0001284 0.54095 0.023468 0.010046 Hippocampus response to injury Chia et al. (2008)
Top2a 1.1833 0.00011062 1.4402 2.84E-05 0.0081051 Cell cycle regulator Gómez-Herreros et al. (2014)
Eomes 1.1415 0.008134 2.38 1.88E-05 0.00023908 Hippocampal progenitor cell marker Nelson et al. (2020)
Zic4 1.099 0.01369 0.91747 0.042865 0.04725 Activator of stem cell proliferation Blank et al. (2011)
Pole 1.0115 0.0000071 0.34811 0.039086 0.011691 Cell cycle regulator Hogg and Johansson (2012)
Insm2 0.99643 0.0470330 1.1039 0.044601 0.02797 Glycolitic pathway activator Wang et al. (2018)
Mocos 0.94776 0.0000387 0.3783 0.020257 0.015645 Negative ROS regulator Féron et al. (2016)
Unc13d 0.89326 0.0000370 0.46491 0.024812 0.025937 Synaptic plasticity regulator Dittman (2019)
Slc18a2 0.88805 0.0012173 0.86778 0.0011748 0.016933 Synaptic plasticity regulator Stefanovic et al. (2016)
Prc1 0.87277 0.01267 1.1565 0.0027045 0.0083644 Cell cycle regulator Mollinari et al. (2002)
Cenpf 0.82583 0.00035633 0.8323 0.00013571 0.0014211 Synaptic plasticity regulator Tanaka et al. (2012)
Gstm2 0.77843 0.0048713 0.81297 0.0047004 0.021936 Negative ROS regulator Segura-Aguilar et al. (2014)
Dpf3 0.44617 0.000012 0.28425 0.0014714 0.0006557 Promotes stem cell proliferation Lessard et al. (2007)
Trp73 0.41055 0.04942 0.31332 0.027558 0.018847 Promotes stem cell proliferation Talos et al. (2010)
Ncapd2 0.38713 0.0001085 0.16293 0.04469 0.023229 Promotes cell proliferation He et al. (2023)
Lpin2 0.37591 8.83E-11 0.17873 0.015751 0.00075486 Fatty acid metabolism regulator Chen et al. (2015)
Dbn1 0.37119 0.000000031 0.14431 0.010052 0.0081834 Synaptic plasticity regulator Jung et al. (2015), Aoki et al. (2005)
Grm1 0.30756 0.00023736 0.20714 0.0025639 0.0018544 Synaptic plasticity regulator Baskys et al. (2005)
Cog1 0.26607 0.00024705 0.099728 0.047439 0.0038386 Regulator of neuronal vesicles Climer et al. (2018)
Zfp143 0.26463 0.0049829 0.18189 0.043793 0.0011626 Regulator of stemness Choi et al. (2012)
Gabbr2 0.22451 0.000032 0.13118 0.024323 0.0091136 Synaptic plasticity regulator Heaney and Kinney (2016)
Glrb −0.14419 0.036066 −0.14323 0.0083886 0.0010861 Synaptic plasticity regulator Lee et al. (2017)
Cnbp −0.22151 0.0021957 −0.14838 0.0061765 0.031318 Proliferation regulator Calcaterra et al. (2010)
Nrxn1 −0.23221 0.00000080 −0.10842 0.013282 0.013987 Synaptic plasticity regulator Luo et al. (2021)
Mzt1 −0.2335 0.00020106 −0.15501 0.014654 2.71E-06 Cell division regulator Batzenschlager et al. (2014)
Purb −0.24099 0.00065256 −0.1747 0.00039673 2.41E-05 Exercise-dependent synaptic plasticity regulator Chen et al. (2007)
Fgf12 −0.25472 0.00087818 −0.15989 0.01975 0.032654 Synaptic plasticity regulator Zhang et al. (2012)
Pbx1 −0.26346 0.0000677 −0.1229 0.016314 0.043764 Stem cell regulator Hau et al. (2021)
Nudt3 −0.28487 0.0015394 −0.17299 0.032189 0.013985 Involved in insulin signaling Williams et al. (2015)
Pou3f3 −0.31284 0.0000008 −0.17004 0.038589 0.00028684 Stem cell regulator; favors quiescence Castro et al. (2006)
Galk1 −0.34371 0.0032957 −0.26377 0.041561 0.0028659 Glycolysis regulator Oh et al. (2020)
Crim1 −0.3589 0.0016505 −0.24056 0.013359 0.00066872 Involved in neural development Ponferrada et al. (2012)
Rgs14 −0.38356 0.0000044 −0.22733 0.0043943 8.33E-05 Synaptic plasticity negative regulator Evans et al. (2018)
Grm3 −0.50434 0.00086289 −0.15248 0.045464 0.020496 Synaptic plasticity regulator, Neurotransmitter receptor Dogra et al. (2021)
Ttc9 −0.5047 0.0000805 −0.33005 0.01287 0.002817 Cell migration ? Cao et al. (2008)
Klhl4 −0.50857 0.0017137 −0.31945 0.02117 0.0069358 Cell cycle regulator Choi et al. (2020)
Gabra3 −0.595 0.0017071 −0.2893 0.041659 0.015167 Synaptic plasticity regulator, Neurotransmitter receptor Wagner et al. (2021)
Ramp3 −0.75442 0.0010749 −0.60432 0.018916 0.015335 Pro-differentiative Li et al. (2021b)
Sp8 −1.0514 0.0048701 −0.88825 0.022895 0.012781 Marker of quiescent stem cells Zhang et al. (2016)
Chrm5 −1.3817 0.010806 −1.0349 0.025201 0.026236 Synaptic plasticity regulator, Neurotransmitter receptor Sakata and Overacre (2017)
Nov −1.6364 0.034359 −0.85695 0.0011469 0.016149 Inhibitor of progenitor cell proliferation Le Dréau et al. (2010)