Accompanying with companion ameliorated CUMS-induced depression-like behaviors in mice.
In order to identify the effect of rewards on ameliorating depression-like behaviors, as a classic depression model [49], CUMS model was used to explore the possible antidepressant effect of accompanying with companion. As shown in Fig. 1a, C57BL/6J male mice were treated with CUMS or CUMS-companion for 4 weeks, after which depression-like behaviors or resilience were assessed by sucrose preference test, Y-maze test and forced swimming test. According to the behavioral tests, only mice changed significantly in all three tests could they be defined as MDD mice, and mice did not change significantly in all three tests could they be defined as resilience mice (Fig. 1a). The results showed that three behavioral parameters of the CUMS and CUMS-companion groups varied so much after 4 weeks treatments (Fig. 1b-d). In CUMS group mice, the SPT values significantly decreased after 4 weeks treatments (83.86 ± 0.9225% versus 71.51 ± 1.788%, p < 0.0001), n = 67)(Figure 1b and Table S1), the ratios of stay time in M-arm to stay time in total arms also showed significant difference after treatments (78.54 ± 1.177% versus 57.22 ± 2.05%, p < 0.0001, n = 67) (Fig. 1c and Table S2), and the FST’s immobile time were 206.6 ± 7.086 seconds after treatments and 137 ± 6.935 seconds before the CUMS treatments (p < 0.0001, n = 67) (Fig. 1d and Table S3). These results indicated that CUMS treatments can induce depression-like behaviors in mice.
It is noteworthy that pairwise comparisons between the three groups after CUMS showed significant differences in SPT, YMT and FST, respectively (Fig. 1b-d, Table S1-3). Compared with CUMS group, the SPT and YMT values significantly increased in CUMS-companion group, and the ratios of stay time in M-arm to stay time in total arms showed significantly decrease in CUMS-companion group (Fig. 1b-d, Table S1-3). In addition, as indicated in Table 1, mice treated by the CUMS or CUMS and companion in 4 weeks, the percentage of MDD in CUMS group was about 29.85%, while that in CUMS-companion group decreased to 12.12%. Meanwhile, the percentage of resilience in CUMS group was about 11.94%, while that in CUMS-companion group increased to 36.36%. These results suggested that accompanying with companion ameliorated CUMS-induced depression-like behaviors in mice.
Table 1
Behavioral changes in the group of CUMS and CUMS-companion mice.
Mice | CUMS | Percentage (%) | CUMS-companion | Percentages (%) |
MDD | 20 | 29.85 | 8 | 12.12 |
Resilience | 8 | 11.94 | 24 | 36.36 |
Atypical | 39 | 58.21 | 34 | 51.52 |
Total | 67 | 100 | 66 | 100 |
Accompanying with companion disturbed the mRNA expression of medial prefrontal cortex in CUMS-induced MDD and resilience mice.
To investigate the molecular mechanism of ameliorated effect of accompanying with companion in CUMS-induced depression-like behaviors, the miRNA and mRNA profiles were analyzed by high-throughput sequencing in medial prefrontal harvested from control, CUMS-MDD, Reward-MDD, CUMS-resilience, Reward-resilience mice. 15 samples were sequenced using RNA-Seq technology, and the number of raw sequencing reads and clean reads are shown in Supplementary Table S4. The unique mapping ratio with reference gene and the average genome mapping ratio were more than 79.48% (Supplementary Table S4). The criterion to make sure differential expression of genes were a 1.5-fold or greater change in transcript level between any two group mice and a P-value < 0.05. As shown in Supplementary Table S5, 311 differentially expressed mRNAs were obtained in control versus CUMS-MDD mice, in which 195 mRNAs were up-regulated and 116 mRNAs were down-regulated in CUMS-MDD mice compared to control mice. In the control versus Reward-MDD mice, 127 differentially expressed mRNAs changed in Reward-MDD mice compared to control mice, where 21 mRNAs were down-regulated and 106 mRNAs were up-regulated. For resilience mice derived from two treatments, 45 differentially expressed mRNAs were obtained in control versus CUMS-resilience mice, in which 24 mRNAs are upregulated and 21 mRNAs are downregulated; 45 differentially expressed mRNAs were obtained in control versus Reward-resilience mice, in which 22 mRNAs are upregulated and 23 mRNAs are downregulated. These results indicate that the molecular changes in the medial prefrontal region are different in different treatments that achieve consistent behaviors.
In order to validate the sequencing data above, we ran quantitative RT-PCR (qRT-PCR) from tissues that were used for mRNA sequencing. As shown in Supplementary Figure S1-S3, the expressions of Ccl28, Ciart, Fkbp5, Fmo2, Gm7120, Gpr149, Hif3a, Kcnh5, Npas4, Plin4, Xdh were decreased, as well as the expressions of Slc6a13 was increased in Reward-MDD mice, compared to control mice (Figure S1). Moreover, the expressions of Ciart, Dbp, Fmo2, Hif3a, Lrrc39, Xdh were decreased, as well as the expressions of Col3a1, Pcdha1, Slc6a13 and Stac were increased in Reward-resilience mice, compared to control mice (Figure S2). Consistent results achieved by mRNA sequencing and qRT-PCR confirm the validation of our study.
Identification of differentially expressed genes are associated with depression-like behaviors.
Since the depression-like behaviors in MDD mice were consistent whether they were derived from CUMS treated group or CUMS-companion group, the common differentially expressed genes (DEGs) in control versus CUMS-MDD and control versus Reward-MDD comparisons may be strongly associated with depression-like behaviors. To identify both unique and common genes in control versus CUMS-MDD and control versus Reward-MDD comparisons, numbers were calculated and presented using a Venn diagram (Fig. 2). As shown in Fig. 2 and Supplementary Table S6, 266 DEGs (I) specifically involved in control versus CUMS-MDD comparison, and 82 unique DEGs (III) in control versus Reward-MDD comparison. In addition, we obtained 45 common DEGs (II) in control versus CUMS-MDD and control versus Reward-MDD comparisons, such as Anln, Ano2, Atp10a, Ccdc152, Ccdc187, Ccl28, Cd44, Cdh19, Clic5, Cpm, Crb1, Ecm1, Etnppl, Fmo2, Fmod, Gm14440, Gm15446, Gm21949, Gm7120, Gpt, Grin1os, Hapln2, Hhip, Hif3a, Htr4, Il12rb1, Kdr, Mertk, Nptx2, Pcdha4, Plin4, Rcn3, Sdc4, Sec14l5, Slc6a13, Slco4a1, Smad6, Smim3, St18, Tmem88, Tsc22d3, Ugt8a, Wdr86, Xdh and 1700047M11Rik. More importantly, the expression pattern of each common gene was consistent in both CUMS-MDD and Reward-MDD mice compared to control mice, either the expression simultaneously increased or decreased.
To gain in-depth insights into the molecular function of these unique and common DEGs, we performed Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. As shown in Fig. 2 and Supplementary Table S6, 266 unique DEGs (I) were found to be mainly enriched in KEGG pathways associated with Metabolic pathways, PI3K-Akt signaling pathway, Wnt signaling pathway, Hippo signaling pathway, Cytokine-cytokine receptor interaction, ECM-receptor interaction, Neuroactive ligand-receptor interaction, MAPK signaling pathway, Axon guidance and Inflammatory mediator regulation of TRP channels. 82 unique DEGs (III) were found to be mainly enriched in KEGG pathways associated with Metabolic pathways, MAPK signaling pathway, ECM-receptor interaction, Serotonergic synapse, Dopaminergic synapse and Neuroactive ligand-receptor interaction. What is more, 45 common DEGs (II) were mainly enriched in KEGG pathways associated with Metabolic pathways, cAMP signaling pathway, ECM-receptor interaction, PI3K-Akt signaling pathway, Serotonergic synapse and Neuroactive ligand-receptor interaction.
Identification of differentially expressed genes are associated with resilience.
Similar to the depression-like behaviors of MDD mice, the behaviors of resilience mice derived from CUMS treated group and CUMS-companion group were also consistent. Therefore, the common differentially expressed genes (DEGs) are associated with resilience were identified, calculated and presented using a Venn diagram (Fig. 3). As shown in Fig. 3 and Supplementary Table S7, 37 DEGs (I) specifically involved in control versus CUMS-resilience comparison, and 37 unique DEGs (III) in control versus Reward-resilience comparison. In addition, we obtained 8 common DEGs (II) in control versus CUMS-resilience and control versus Reward-resilience comparisons, such as Cpm, Fmo2, Grin1os, Hif3a, Hist2h2aa2, Plin4, Scn5a and Trex1. What is more, the expression pattern of each common gene was consistent in both CUMS-resilience and Reward-resilience mice compared to control mice, either the expression simultaneously increased or decreased. Compared with MDD mice, the resilience mice derived from CUMS treated group and CUMS-companion group seem to share less molecular changes.
Subsequently, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis was performed for unique DEGs (I), unique DEGs (III) and common DEGs (II). As shown in Fig. 3 and Supplementary Table S7, 37 unique DEGs (I) were found to be mainly enriched in KEGG pathways associated with cAMP signaling pathway, Neuroactive ligand-receptor interaction, Metabolic pathways, PI3K-Akt signaling pathway, Wnt signaling pathway, ECM-receptor interaction, Regulation of actin cytoskeleton and Tight junction. 37 unique DEGs (III) were found to be mainly enriched in KEGG pathways associated with ECM-receptor interaction, PI3K-Akt signaling pathway, Metabolic pathways, p53 signaling pathway, Alcoholism, Caffeine metabolism, Biosynthesis of amino acids and Rap1 signaling pathway. What is more, 8 common DEGs (II) were mainly enriched in KEGG pathways associated with Alcoholism and Adrenergic signaling in cardiomyocytes.
Identification of differentially expressed genes are associated with accompanying with companion.
Most individuals may not suffer from major depression by chronic stress, i.e., resilience [50]. As shown in Table 1, approximately 11.94% of CUMS-treated mice in 4 weeks were resilience, and this value increased to approximately 36.36% after accompanying with companion. To identify the DEGs are associated with the accompanying with companion treatments, differentially expressed mRNAs were screened in CUMS-resilience versus Reward-resilience mice. As shown in Supplementary Table S5, 59 differentially expressed mRNAs were obtained in CUMS-resilience versus Reward-resilience mice, in which 26 mRNAs were up-regulated and 33 mRNAs were down-regulated in CUMS-MDD mice compared to control mice. To gain in-depth insights into the molecular function of these differentially expressed mRNAs, we performed KEGG pathway enrichment analysis. As shown in Table 2, differentially expressed mRNAs were found to be mainly enriched in KEGG pathways associated with Neuroactive ligand-receptor interaction, Dopaminergic synapse, Rap1 signaling pathway, Focal adhesion, Metabolic pathways and PI3K-Akt signaling pathway.
Table 2
Signaling pathways identified by KEGG function analysis based on DEGs data in CUMS-resilience versus Reward-resilience.
KEGG Entry | Term | Count | Genes |
mmu04510 | Focal adhesion | 3 | Rac3 (RAS-related C3 botulinum substrate 3) ↓, Col6a3 (collagen, type VI, alpha 3) ↓, Thbs1 (thrombospondin 1) ↑ |
mmu05219 | Bladder cancer | 2 | Mmp9 (matrix metallopeptidase 9) ↓, Thbs1 (thrombospondin 1) ↑ |
mmu04015 | Rap1 signaling pathway | 3 | Rac3 (RAS-related C3 botulinum substrate 3) ↓, Drd2 (dopamine receptor D2) ↓, Thbs1 (thrombospondin 1) ↑ |
mmu04115 | p53 signaling pathway | 2 | Sfn (stratifin) ↓, Thbs1 (thrombospondin 1) ↑ |
mmu04512 | ECM-receptor interaction | 2 | Col6a3 (collagen, type VI, alpha 3) ↓, Thbs1 (thrombospondin 1) ↑ |
mmu04974 | Protein digestion and absorption | 2 | Col9a2 (collagen, type IX, alpha 2) ↑, Col6a3 (collagen, type VI, alpha 3) ↓ |
mmu04728 | Dopaminergic synapse | 2 | Drd2 (dopamine receptor D2) ↓, Drd4 (dopamine receptor D4) ↓ |
mmu04550 | Signaling pathways regulating pluripotency of stem cells | 2 | Lhx5 (LIM homeobox protein 5) ↑, Klf4 (Kruppel-like factor 4) ↑ |
mmu04024 | cAMP signaling pathway | 2 | Rac3 (RAS-related C3 botulinum substrate 3) ↓, Drd2 (dopamine receptor D2) ↓ |
mmu05205 | Proteoglycans in cancer | 2 | Mmp9 (matrix metallopeptidase 9) ↓, Thbs1 (thrombospondin 1) ↑ |
mmu05206 | MicroRNAs in cancer | 2 | Mmp9 (matrix metallopeptidase 9) ↓, Thbs1 (thrombospondin 1) ↑ |
mmu04080 | Neuroactive ligand-receptor interaction | 2 | Drd2 (dopamine receptor D2) ↓, Drd4 (dopamine receptor D4) ↓ |
mmu04151 | PI3K-Akt signaling pathway | 2 | Col6a3 (collagen, type VI, alpha 3) ↓, Thbs1 (thrombospondin 1) ↑ |
mmu05200 | Pathways in cancer | 2 | Rac3 (RAS-related C3 botulinum substrate 3) ↓, Mmp9 (matrix metallopeptidase 9) ↓ |
mmu01100 | Metabolic pathways | 3 | Aldh1a2 (aldehyde dehydrogenase family 1, subfamily A2) ↑, Alas2 (aminolevulinic acid synthase 2, erythroid) ↓, Gpt (glutamic pyruvic transaminase, soluble) ↓ |
mmu04919 | Thyroid hormone signaling pathway | 1 | Dio3 (deiodinase, iodothyronine type III) ↑ |
mmu04931 | Insulin resistance | 1 | Nr1h3 (nuclear receptor subfamily 1, group H, member 3) ↑ |
mmu00220 | Arginine biosynthesis | 1 | Gpt (glutamic pyruvic transaminase, soluble) ↓ |
mmu01200 | Carbon metabolism | 1 | Gpt (glutamic pyruvic transaminase, soluble) ↓ |
mmu04662 | B cell receptor signaling pathway | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu00250 | Alanine, aspartate and glutamate metabolism | 1 | Gpt (glutamic pyruvic transaminase, soluble) ↓ |
mmu05010 | Alzheimer's disease | 1 | Atp2a1 (ATPase, Ca + + transporting, cardiac muscle, fast twitch 1) ↓ |
mmu04360 | Axon guidance | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu04810 | Regulation of actin cytoskeleton | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu05212 | Pancreatic cancer | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu00260 | Glycine, serine and threonine metabolism | 1 | Alas2 (aminolevulinic acid synthase 2, erythroid) ↓ |
mmu04932 | Non-alcoholic fatty liver disease (NAFLD) | 1 | Nr1h3 (nuclear receptor subfamily 1, group H, member 3) ↑ |
mmu04310 | Wnt signaling pathway | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu05160 | Hepatitis C | 1 | Nr1h3 (nuclear receptor subfamily 1, group H, member 3) ↑ |
mmu03010 | Ribosome | 1 | Rpl37rt (ribosomal protein L37, retrotransposed) ↓ |
mmu00830 | Retinol metabolism | 1 | Aldh1a2 (aldehyde dehydrogenase family 1, subfamily A2) ↑ |
mmu05412 | Arrhythmogenic right ventricular cardiomyopathy (ARVC) | 1 | Pkp2 (plakophilin 2) ↓ |
mmu04022 | cGMP-PKG signaling pathway | 1 | Atp2a1 (ATPase, Ca + + transporting, cardiac muscle, fast twitch 1) ↓ |
mmu04664 | Fc epsilon RI signaling pathway | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu05202 | Transcriptional misregulation in cancer | 1 | Mmp9 (matrix metallopeptidase 9) ↓ |
mmu01230 | Biosynthesis of amino acids | 1 | Gpt (glutamic pyruvic transaminase, soluble) ↓ |
mmu04370 | VEGF signaling pathway | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu05161 | Hepatitis B | 1 | Mmp9 (matrix metallopeptidase 9) ↓ |
mmu05144 | Malaria | 1 | Thbs1 (thrombospondin 1) ↑ |
mmu04350 | TGF-beta signaling pathway | 1 | Thbs1 (thrombospondin 1) ↑ |
mmu05231 | Choline metabolism in cancer | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu04621 | NOD-like receptor signaling pathway | 1 | Pstpip1 (proline-serine-threonine phosphatase-interacting protein 1) ↓ |
mmu04960 | Aldosterone-regulated sodium reabsorption | 1 | Sfn (stratifin) ↓ |
mmu05416 | Viral myocarditis | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu00860 | Porphyrin and chlorophyll metabolism | 1 | Alas2 (aminolevulinic acid synthase 2, erythroid) ↓ |
mmu04540 | Gap junction | 1 | Drd2 (dopamine receptor D2) ↓ |
mmu04145 | Phagosome | 1 | Thbs1 (thrombospondin 1) ↑ |
mmu04020 | Calcium signaling pathway | 1 | Atp2a1 (ATPase, Ca + + transporting, cardiac muscle, fast twitch 1) ↓ |
mmu04520 | Adherens junction | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu05012 | Parkinson's disease | 1 | Drd2 (dopamine receptor D2) ↓ |
mmu04650 | Natural killer cell mediated cytotoxicity | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu04668 | TNF signaling pathway | 1 | Mmp9 (matrix metallopeptidase 9) ↓ |
mmu04670 | Leukocyte transendothelial migration | 1 | Mmp9 (matrix metallopeptidase 9) ↓ |
mmu04110 | Cell cycle | 1 | Sfn (stratifin) ↓ |
mmu04514 | Cell adhesion molecules (CAMs) | 1 | Mpz (myelin protein zero) ↑ |
mmu04972 | Pancreatic secretion | 1 | Atp2a1 (ATPase, Ca + + transporting, cardiac muscle, fast twitch 1) ↓ |
mmu01210 | 2-Oxocarboxylic acid metabolism | 1 | Gpt (glutamic pyruvic transaminase, soluble) ↓ |
mmu04071 | Sphingolipid signaling pathway | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu05030 | Cocaine addiction | 1 | Drd2 (dopamine receptor D2) ↓ |
mmu03320 | PPAR signaling pathway | 1 | Nr1h3 (nuclear receptor subfamily 1, group H, member 3) ↑ |
mmu04010 | MAPK signaling pathway | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu04014 | Ras signaling pathway | 1 | Rac3 (RAS-related C3 botulinum substrate 3) ↓ |
mmu04915 | Estrogen signaling pathway | 1 | Mmp9 (matrix metallopeptidase 9) ↓ |
mmu05034 | Alcoholism | | Drd2 (dopamine receptor D2) ↓ |
Note: ↑ indicates up-regulation in the tissue of mPFC from Reward-resilience mice compared to CUMS-resilience mice, whereas ↓ represents down-regulation. |
Furthermore, to validate the sequencing data above, we ran quantitative RT-PCR (qRT-PCR) from tissues that were used for mRNA sequencing. 10 mRNAs were selected to conduct qRT-PCR, the expressions of Artn, Dio3, Klf4, Pcdha1, Pcdhgb4, and Slc6a13 are increased, as well as the expressions of Col6a3, Drd2, Mmp9, and Rac3 are decreased (Figure S3). Consistent results achieved by mRNA sequencing and qRT-PCR confirm the validation of our study.
Accompanying with companion altered miRNA expression in resilience mice.
Studies have revealed that patients with psychiatric disorders have altered miRNA expression profiles in the brain and circulation, and multiple roles of miRNAs in psychiatric disorders have summarized [51]. To investigate the impact of accompanying with companion on miRNA, we performed differential expression analysis of miRNA by high-throughput sequencing. As shown in Table S8, compared with the CUMS-resilience mice, a total of 61 differentially expressed miRNAs were up-regulated and 135 differentially expressed miRNAs were down-regulated in the Reward-resilience mice. In order to validate the finding by sequencing miRNA analysis, quantitative RT-PCR were performed for certain miRNAs. As shown in Figure S4, qRT-PCR analysis results for miR-149-5p, miR-183-5p, miR-199b-3p, miR-323-5p, miR-337-3p, miR-384-5p, miR-451a, miR-484, miR-671-5p, and miR-702-3p were consistent with miRNA sequencing results in Reward-resilience versus CUMS-resilience mice.
miRNAs can act as an ‘expression switch’ and block the expression of their target genes [52]. In this study,if the mRNAs downregulated were caused by miRNAs, their correspondent miRNAs should be upregulated, or vice versa. Based on three databases (RNAhybrid, Targetscan and miRanda), the target genes of miRNAs were predicted and matched with the mRNA measured by mRNA sequencing to form the complex interactions between miRNAs and mRNAs (Tables 3 and 4). As shown in Fig. 4a and 4b, 63 miRNAs established regulatory relationships with 34 mRNAs in CUMS-resilience versus Reward-resilience mice. Furthermore, the miRNAs targeted mRNAs were validated by qRT-PCR and dual luciferase reporter assay. As shown in Fig. 4c and 4d, there were inverse correlations between Pcdha1 and miRNA-337-3p, as well as between Rac3 and miRNA-671-5p from qRT-PCR analysis. In dual luciferase report assay, the relative activities of luciferase reporter for Pcdha1 and Rac3 are significantly decreased by the mimics of miRNA-337-3p and miRNA-671-5p respectively, but not negative control (Fig. 4e and 4f). These reductions can be reversed by changing the binding sites of miRNA-337-3p and miRNA-671-5p in Pcdha1 and Rac3. These results support that mRNA Pcdha1 is the direct target of miRNA-337-3p, and mRNA Rac3 is the direct target of miRNA-671-5p, which is consistent with our bioinformatics analyses in the prediction of miRNA target genes.
Table 3
Differently expression miRNAs predict target mRNAs in CUMS-resilience versus Reward-resilience.
miRNAs | The predicted target mRNAs that match DEGs in transcriptome * | miRNAs | The predicted target mRNAs that match DEGs in transcriptome * |
miR-1231-3p↓ | Aldh1a2↑, Klf4↑ | miR-5617-3p↓ | Cyr61↑ |
miR-1249-5p↓ | Alx3↑, Mpz↑ | miR-652-5p↓ | Crispld2↑, Nr1h3↑ |
miR-1291↓ | Adam33↑ | miR-6538↓ | Alx3↑, Artn↑, Dio3↑, Hist1h1c↑ |
miR-141-5p↑ | Obscn↓ | miR-669m-5p↑ | Obscn↓ |
miR-146b-3p↓ | Adam33↑ | miR-670-5p↑ | Obscn↓ |
miR-149-5p↓ | Aldh1a2↑, Col9a2↑, Lhx5↑, Pcdhgb4↑ | miR-671-5p↑ | Alas2↓, Atp2a1↓, Col6a3↓, Efcab6↓, Mmp9↓, Rac3↓ |
miR-1931↓ | Plk5↑ | miR-6919-3p↓ | Adam33↑, Crispld2↑, Nr1h3↑, Pcdha1↑, Plk5↑ |
miR-1934-5p↓ | Adamts1↑ | miR-6919-5p↑ | Col6a3↓, Obscn↓, Pkp2↓ |
miR-196a-5p↓ | Crispld2↑ | miR-6954-5p↓ | Crispld2↑, Hist1h1c↑, Nr1h3↑, Pcdhgb4↑, Thbs1↑ |
miR-196b-5p↓ | Crispld2↑ | miR-6982-5p↓ | Col9a2↑, Mpz↑, Pcdhgb4↑, Rasl11a↑ |
miR-214-3p↓ | Alx3↑, Dio3↑, Plk5↑ | miR-698-5p↓ | Crispld2↑ |
miR-24-3p↓ | Lhx5↑ | miR-6997-5p↓ | Pcdha1↑ |
miR-26b-3p↓ | Slc6a13↑ | miR-6999-5p↑ | Pik3r6↓ |
miR-299a-3p↑ | Col6a3↓ | miR-7004-5p↓ | Adamts1↓ |
miR-3078-5p↓ | Mpz↑ | miR-7019-3p↓ | Col9a2↑, Pcdha1↑, Slc6a13↑ |
miR-3087-5p↓ | Adam33↑, Alx3↑, Mpz↑, Pcdha1↑ | miR-702-3p↓ | Col9a2↑, Dio3↑, Pcdha1↑ |
miR-3093-5p↓ | Crispld2↑, Hist1h1c↑, Plk5↑ | miR-7032-5p↓ | Alx3↑, Artn↑, Pcdha1↑, Pcdhgb4↑, Thbs1↑ |
miR-323-5p↓ | Artn↑, Klf4↑ | miR-7043-3p↑ | Obscn↓ |
miR-328-5p↓ | Alx3↑, Crispld2↑, Hist1h1c↑ | miR-7052-3p↓ | Dio3↑, Lhx5↑ |
miR-337-3p↓ | Nr1h3↑, Pcdha1↑, Syce1l↑ | miR-7053-3p↓ | Crispld2↑ |
miR-339-5p↓ | Alx3↑, Col9a2↑, Pcdhgb4↑ | miR-7054-3p↓ | Lhx5↑ |
miR-341-5p↓ | Aldh1a2↑ | miR-7056-3p↓ | Col9a2↑ |
miR-3544-3p↓ | Adam33↑, Artn↑, Syce1l↑ | miR-7070-3p↓ | Col9a2↑, Nr1h3↑, Pcdha1↑ |
miR-378d↑ | Col6a3↓, Obscn↓, Pkp2↓ | miR-7071-5p↓ | Alx3↑, Artn↑, Col9a2↑, Mpz↑, Rasl11a↑ |
miR-449c-5p↑ | Obscn↓ | miR-7115-5p↓ | Artn↑, Cyr61↑, Hist1h1c↑, Col9a2↑, Pcdhgb4↑ |
miR-466 h-5p↑ | Obscn↓, Lrrc10b↓ | miR-7235-3p↑ | Efcab3↑ |
miR-484↓ | Col9a2↑ | miR-758-5p↓ | Col9a2↑ |
miR-486b-5p↑ | 4933409K07Rik↓ | miR-760-3p↓ | Adam33↑, Alx3↑, Col9a2↑, Crispld2↑, Nr1h3↑ |
miR-503-5p↑ | Efcab6↓, Obscn↓ | miR-770-3p↓ | Adam33↑ |
miR-5126↓ | Alx3↑, Dio3↑, Klf4↑ | miR-873a-5p↑ | Obscn↓ |
miR-5129-5p↑ | Obscn↓ | miR-873a-5p↑ | Tmem40↓ |
Note: ↑ indicates up-regulation in the tissue of mPFC from Reward-resilience mice compared to CUMS-resilience mice, whereas ↓ represents down-regulation. |
Table 4
Differently expression mRNAs predict target miRNAs in CUMS-resilience versus Reward-resilience.
Gene Symbol | The predicted target miRNAs that match DEGs in transcriptome * | Gene Symbol | The predicted target miRNAs that match DEGs in transcriptome * |
Adam33↑ | miR-1291↓, miR-146b-3p↓, miR-3087-5p↓, miR-3544-3p↓, miR-6919-3p↓, miR-760-3p↓, miR-770-3p↓ | Lrrc10b↓ | miR-466p-3p↑ |
Adamts1↑ | miR-1934-5p↓, miR-7004-5p↓ | Mmp9↓ | miR-671-5p↑ |
Alas2↓ | miR-671-5p↑ | Mpz↑ | miR-1249-5p↓, mmiR-3078-5p↓, miR-3087-5p↓, miR-6982-5p↓, miR-7071-5p↓ |
Aldh1a2↑ | miR-1231-3p↓, miR-149-5p↓, miR-341-5p↓ | Nr1h3↑ | miR-337-3p↓, miR-652-5p↓, miR-6919-3p↓, miR-6954-5p↓, miR-7070-3p↓, miR-760-3p↓ |
Alx3↑ | miR-1249-5p↓, miR-214-3p↓, miR-3087-5p↓, miR-328-5p↓, miR-339-5p↓, miR-5126↓, miR-6538↓, miR-7032-5p↓, miR-7071-5p↓, miR-760-3p↓ | Obscn↓ | miR-141-5p↑, miR-378d↑, miR-449c-5p↑, miR-466 h-5p↑, miR-503-5p↑, miR-5129-5p↑, miR-669m-5p↑, miR-670-5p↑, miR-6919-5p↑, miR-7043-3p↑,miR-873a-5p↑ |
Artn↑ | miR-323-5p↓, miR-3544-3p↓, miR-6538↓, miR-7032-5p↓, mmu-miR-7071-5p↓, miR-7115-5p↓ | Pcdha1↑ | miR-3087-5p↓, miR-337-3p↓, miR-6919-3p↓, miR-6997-5p↓, miR-7019-3p↓, miR-702-3p↓, miR-7032-5p↓, miR-7070-3p↓ |
Atp2a1↓ | miR-671-5p↑ | Pcdhgb4↑ | miR-149-5p↓, miR-339-5p↓, miR-6954-5p↓, miR-6982-5p↓, miR-7032-5p↓, miR-7117-5p↓ |
Col6a3↓ | miR-299a-3p↑, miR-378d↑, miR-671-5p↑, miR-6919-5p↑ | Pik3r6↓ | miR-6999-5p↑ |
Col9a2↑ | miR-149-5p↓, miR-339-5p↓, miR-484↓, miR-6982-5p↓, miR-7019-3p↓, -miR-702-3p↓, miR-702-3p↓, miR-7056-3p↓, miR-7070-3p↓, miR-7071-5p↓, miR-7117-5p↓, miR-758-5p↓, miR-760-3p↓ | Pkp2↓ | miR-378d↑, miR-6919-5p↑ |
Crispld2↑ | miR-196a-5p↓, miR-196b-5p↓, miR-3093-5p↓, miR-328-5p↓, miR-652-5p↓, miR-6919-3p↓, miR-6954-5p↓, miR-698-5p↓, miR-7053-3p↓, miR-760-3p↓ | Plk5↑ | miR-1931↓, miR-214-3p↓, miR-3093-5p↓, miR-6919-3p↓ |
Cyr61↑ | miR-5617-3p↓, miR-7115-5p↓ | Rac3↓ | miR-671-5p↑ |
Dio3↑ | miR-214-3p↓, miR-5126↓, miR-6538↓, miR-702-3p↓, miR-7052-3p↓ | Rasl11a↑ | miR-6982-5p↓, miR-7071-5p↓ |
Efcab3↓ | miR-7235-3p↑ | Slc6a13↑ | miR-26b-3p↓, miR-7019-3p↓ |
Efcab6↓ | miR-503-5p↑, miR-671-5p↑ | Syce1l↑ | miR-337-3p↓, miR-3544-3p↓ |
Hist1h1c↑ | miR-3093-5p↓, miR-328-5p↓, miR-6538↓, miR-6954-5p↓, miR-7115-5p↓ | Thbs1↑ | miR-6954-5p↓, miR-7032-5p↓ |
Klf4↑ | miR-1231-3p↓, miR-323-5p↓, miR-5126↓ | Tmem40↓ | miR-873a-5p↑ |
Lhx5↑ | miR-149-5p↓, miR-24-3p↓, miR-7052-3p↓, miR-7054-3p↓ | | |
Note: ↑ indicates up-regulation in the tissue of mPFC from Reward-resilience mice compared to CUMS-resilience mice, whereas ↓ represents down-regulation. |