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. DOI: 10.12300/j.issn.1674-5817.2023.006.

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

References 23

1	CARLI M, KOLACHALAM S, LONGONI B, et al. Atypical antipsychotics and metabolic syndrome: from molecular mechanisms to clinical differences[J]. Pharmaceuticals, 2021, 14(3):238. DOI: 10.3390/ph14030238 .
2	LORD C C, WYLER S C, WAN R, et al. The atypical antipsychotic olanzapine causes weight gain by targeting serotonin receptor 2C[J]. J Clin Invest, 2017, 127(9):3402-3406. DOI: 10.1172/JCI93362 .
3	ANDERMANN M L, LOWELL B B. Toward a wiring diagram understanding of appetite control[J]. Neuron, 2017, 95(4):757-778. DOI: 10.1016/j.neuron.2017.06.014 .
4	STIP E, LUNGU O V, ANSELMO K, et al. Neural changes associated with appetite information processing in schizophrenic patients after 16 weeks of olanzapine treatment[J]. Transl Psychiatry, 2012, 2(6): e128. DOI: 10.1038/tp.2012.53 .
5	KEREM L, LAWSON E A. The effects of oxytocin on appetite regulation, food intake and metabolism in humans[J]. Int J Mol Sci, 2021, 22(14):7737. DOI: 10.3390/ijms22147737 .
6	LAWSON E A, OLSZEWSKI P K, WELLER A, et al. The role of oxytocin in regulation of appetitive behaviour, body weight and glucose homeostasis[J]. J Neuroendocrinol, 2020, 32(4): e12805. DOI: 10.1111/jne.12805 .
7	PLATZER M, FELLENDORF F T, BENGESSER S A, et al. The relationship between food craving, appetite-related hormones and clinical parameters in bipolar disorder[J]. Nutrients, 2020, 13(1):76. DOI: 10.3390/nu13010076 .
8	MATHEWS J, NEWCOMER J W, MATHEWS J R, et al. Neural correlates of weight gain with olanzapine[J]. Arch Gen Psychiatry, 2012, 69(12):1226. DOI: 10.1001/archgenpsychiatry. 2012.934 .
9	BENÍTEZ-ANGELES M, MORALES-LÁZARO S L, JUÁREZ-GONZÁLEZ E, et al. TRPV1: structure, endogenous agonists, and mechanisms[J]. Int J Mol Sci, 2020, 21(10):3421. DOI: 10.3390/ijms21103421 .
10	KOMARNYTSKY S, RATHINASABAPATHY T, WAGNER C, et al. Endocannabinoid system and its regulation by polyunsaturated fatty acids and full spectrum hemp oils[J]. Int J Mol Sci, 2021, 22(11):5479. DOI: 10.3390/ijms22115479 .
11	POTVIN S, LUNGU O V, STIP E. Anandamide is involved in appetite-related amygdala hyperactivations in schizophrenia patients treated with olanzapine: a functional magnetic resonance imaging study[J]. J Clin Psychopharmacol, 2015, 35(1):82-83. DOI: 10.1097/JCP.0000000000000236 .
12	DERBENEV A V, ZSOMBOK A. Potential therapeutic value of TRPV1 and TRPA1 in diabetes mellitus and obesity[J]. Semin Immunopathol, 2016, 38(3):397-406. DOI: 10.1007/s00281-015-0529-x .
13	BENARROCH L, KOWALCHUK C, WILSON V, et al. Atypical antipsychotics and effects on feeding: from mice to men[J]. Psychopharmacology, 2016, 233(14):2629-2653. DOI: 10.1007/s00213-016-4324-8 .
14	刘玉芝, 徐乐平, 袁娇, 等. 奥氮平对精神分裂症患者胃动力学的影响[J]. 中国健康心理学杂志, 2013, 21(8): 1149-1151. DOI: 10.3969/j.issn.1005-4847.2010.05.014 .
	LIU Y Z, XU L P, YUAN J, et al. The influence of olanzapine in the treatment of schizophrenic patients on the gastric motility[J]. China J Health Psychol, 2013, 21(8): 1149-1151. DOI: 10.3969/j.issn.1005-4847.2010.05.014 .
15	FANG H, JUDD R L. Adiponectin regulation and function[J]. Compr Physiol, 2018, 8(3): 1031-1063. DOI: 10.1002/cphy.c170046 .
16	RAMANANTSOA N, MATROT B, VARDON G, et al. Impaired ventilatory and thermoregulatory responses to hypoxic stress in newborn phox2b heterozygous knock-out mice[J]. Front Physiol, 2011, 2:61. DOI: 10.3389/fphys.2011.00061 .
17	THOMAS S A, PALMITER R D. Thermoregulatory and metabolic phenotypes of mice lacking noradrenaline and adrenaline[J]. Nature, 1997, 387(6628):94-97. DOI: 10.1038/387094a0 .
18	JIMENEZ-PREITNER M, BERNEY X, THORENS B. Plac8 is required for white adipocyte differentiation in vitro and cell number control in vivo[J]. PLoS One, 2012, 7(11): e48767. DOI: 10.1371/journal.pone.0048767 .
19	VANMIERLO T, RUTTEN K, VAN VARK- VAN DER ZEE L C, et al. Cerebral accumulation of dietary derivable plant sterols does not interfere with memory and anxiety related behavior in Abcg5^-/- mice[J]. Plant Foods Hum Nutr, 2011, 66(2):149-156. DOI: 10.1007/s11130-011-0219-3 .
20	NELSON D W, GAO Y, YEN M I, et al. Intestine-specific deletion of acyl-CoA: monoacylglycerol acyltransferase (MGAT) 2 protects mice from diet-induced obesity and glucose intolerance[J]. J Biol Chem, 2014, 289(25):17338-17349. DOI: 10.1074/jbc.M114.555961 .
21	ELLIOTT K L, PAVLÍNKOVÁ G, CHIZHIKOV V V, et al. Development in the mammalian auditory system depends on transcription factors[J]. Int J Mol Sci, 2021, 22(8):4189. DOI: 10.3390/ijms22084189 .
22	Aggleton JP, Brown MW, Albasser MM. Contrasting brain activity patterns for item recognition memory and associative recognition memory: insights from immediate-early gene functional imaging [J]. Neuropsychologia, 2012;50(13):3141-3155. DOI: 10.1016/j.neuropsychologia.2012.05.018 .
23	GALLO F T, KATCHE C, MORICI J F, et al. Immediate early genes, memory and psychiatric disorders: focus on c-fos, Egr1 and arc[J]. Front Behav Neurosci, 2018, 12:79. DOI: 10.3389/fnbeh.2018.00079 .

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