Whole-brain Transcriptomic Analysis of Weight Gain Mice induced by Olanzapine

doi:10.12300/j.issn.1674-5817.2023.006

Abstract

Abstract:

Objective The transcriptome sequencing results of brain tissues of olanzapine-treated mice were analyzed to screen out differentially-expressed genes and explore potential targets of atypical antipsychotics leading to body weight gain. Methods Twenty female C57BL/6 mice were randomly divided into control group (Ctrl) and Olanzapine administration group (Olz), which were given saline and Olanzapine solution by gavage, respectively. The whole brain tissues were collected 8 weeks later for Transcriptome sequencing (RNA-Seq). The possible targets of olanzapine-induced body weight gain were identified by the Gene Ontology (GO) functional annotation analysis, the Kyoto Encyclopedia of Genes and Gnomes (KEGG) pathway enrichment analysis, and protein-protein interaction (PPI) network analysis. Differential expression levels of mRNAs were further verified by real-time quantitative fluorescence PCR (RT-qPCR). Results Compared with Ctrl group, 591 differentially expressed genes were screened in Olz group, including 251 up-regulated genes and 340 down-regulated genes. GO analysis showed that differential genes were widely involved in transcriptional process, among which the expression of genes related to the regulation of digestive system and cold-induced thermogenesis were significantly enriched. KEGG analysis showed that differential genes were widely involved in the interaction between neuroactive ligands and receptors, and the differential genes were significantly enriched in oxytocin signaling, fat digestion and absorption, and cholesterol metabolism pathways. RT-qPCR were performed to verify the expression levels of genes enriched in feeding regulation, gastric kinesis, thermogenesis, fat metabolism and other processes (Oxt, Trpv1, Adipoq, Phox2b, Abcg5, Mogat2, Dbh, Plac8 and Neurog1) as well as hub genes in PPI network (Fos, Dusp1 and Egr2), and the results were consistent with the trend of RNA-Seq. Conclusion Olanzapine administration resulted in changes in central feeding regulation, gastrointestinal motility, thermogenesis and other physiological processes in mice, which might be involved in body weight gain induced by olanzapine.

Key words: Olanzapine, Body weight gain, Whole brain, RNA-Seq, Mice

CLC Number:

R-332

Yuan ZHANG, Han LI, Chengfang ZHANG. Whole-brain Transcriptomic Analysis of Weight Gain Mice induced by Olanzapine[J]. Laboratory Animal and Comparative Medicine, 2023, 43(3): 262-270.

Figures/Tables 6

Table 1 Primer sequences of differential genes used in real-time quantitative fluorescence PCR

基因名称	引物序列（5'→3'）
β-Actin	F: GTGCTATGTTGCTCTAGACTTCG
	R: ATGCCACAGGATTCCATACC
Oxt	F: TTGACCAGATGAACGGAGTG
Oxt	R: AGCTACTCGGATACGGGAGA
Trpv1	F: GGGCGAGACTGTCAACAAGA
Trpv1	R: CGGCTCTATTGCTCCCTGAG
Adipoq	F: TGCCCTGTAACTTCTACCCCA
Adipoq	R: GGCAAGTGTCCTCAACTGTGTC
Phox2b	F: TACGCCGCAGTTCCATACAAACTC
Phox2b	R: TCTTTGAGCTGCGCGCTTGTGAAG
Abcg5	F: GTTCCAAGACTGCTTCTC
Abcg5	R: ATGACTGCCTCTACCTTC
Mogat2	F: CCAGGTTGAGAACACCCCTG
Mogat2	R: CTCGATGGGCTTCCCCACTAT
Dbh	F: AATCTGCAGCCTTTGCCTAA
Dbh	R: TTCAGCATCTGCCTCTGTTG
Plac8	F: TGGACTACAAAGACGATGACGA
Plac8	R: ACAAAAGGTCCCACAGAGGC
Neurog1	F: CGATCCCCTTTTCTCCTTTC
Neurog1	R: TGCAGCAACCTAACAAGTGG
Fos	F: TTTCAACGCGGACTACGAGG
Fos	R: GCGCAAAAGTCCTGTGTGTT
Dusp1	F: TGTGAAGCAGAGGAGGAGC
Dusp1	R: ACGCACGGCATGTTGGTC
Egr2	F: TTGACCAGATGAACGGAGTG
Egr2	R: AGCTACTCGGATACGGGAGA

Figure 1 Changes of body weight and food intake induced by olanzapine administrationNote：A, Changes in body weight induced by olanzapine administration; B, Changes in food intake induced by olanzapine administration. n=10 in each group. Ctrl, control group (intragastric injection with saline); Olz, olanzapine group (intragastric injection with olanzapine). Compared with control group, *P<0.05，**P<0.01.

Figure 2 Transcriptomic analysis of differentially expressed genes after olanzapine administration （Volcano map and GO analysis）Note：A， Volcano map of differentially-expressed genes （logarithmic form of screening condition values were taken for the X and Y axes. Each point in the figure represents a differentially expressed gene. Orange， blue and gray indicate the up-regulated， down-regulated and unchanged genes， respectively）. B， GO analysis of differentially expressed genes （differentially-expressed genes-involved molecular functions， cellular environments and biological processes）. C， GO-net analysis of differentially expressed genes.

Figure 3 Transcriptomic analysis of differentially expressed genes after olanzapine administration （KEEG analysis and PPI network analysis）Note：A， KEEG analysis of differentially expressed genes (different colors represent different path groups or network clusters). B, protein-protein interaction (PPI) network analysis (each connection point is labeled with the name of the proteins. Blue indicates the down-regulated genes, and red shows the up-regulated gene. The larger the dot, the more connections there are. The black line represents the regulatory effect between the two genes).

Figure 4 Real-time quantitative fluorescence RCR verification results of differentially expressed genes in the whole brain tissues of mice in the olanzapine infusion group and control groupNote：Ctrl， control group （intragastric injection with saline）； Olz， olanzapine group （intragastric injection with olanzapine）. n= 10 in each group. Compared with the control group， *P<0.05， **P<0.01.

Table 2 Differential genes changes inced by olanzapine and possible metabolic effects

基因	奥氮平给药的影响	可能的效应
Oxt	↓	胃肠道厌食信号减少，享乐性进食增加，产热减少
Trpv1	↓	胃肠道厌食信号减少，享乐性进食增加，产热减少
Adipoq	↓	脂类合成增加、降解减少
Phox2b	↓	产热减少
Abcg5	↓	脂类吸收增加
Mogat2	↓	脂肪吸收的延迟
Dbh	↓	产热减少
Plac8	↑	产热增加
Neurog1	↑	未知

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