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Transcriptome and histopathological changes in mouse brain infected with Neospora caninum.

Nishimura M, Tanaka S, Ihara F, Muroi Y, Yamagishi J, Furuoka H, Suzuki Y, Nishikawa Y - Sci Rep (2015)

Bottom Line: A GOstat analysis predicted that the upregulated genes were involved in the host immune response.Genes whose expression correlated positively and negatively with parasite numbers were involved in the host immune response, and neuronal morphogenesis and lipid metabolic processes, respectively.These results suggest that changes in the gene expression profile associated with neuronal functions as well as immune responses can contribute to the pathogenesis in N. caninum-infected animals.

View Article: PubMed Central - PubMed

Affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido, Japan.

ABSTRACT
Neospora caninum is a protozoan parasite that causes neurological disorders in dogs and cattle. It can cause nonsuppurative meningoencephalitis and a variety of neuronal symptoms are observed, particularly in dogs. However, the pathogenic mechanism, including the relationship between the parasite distribution and the clinical signs, is unclear. In this study, to understand the pathogenic mechanism of neosporosis, parasite distribution and lesions were assessed in the brain of mice infected with N. caninum (strain Nc-1). Host gene expression was also analyzed with RNA sequencing (RNA-Seq). The histopathological lesions in the frontal lobe and the medulla oblongata were significantly more severe in symptomatic mice than in asymptomatic mice, although no association between the severity of the lesions and parasite numbers was found. In infected mice, the expression of 772 mouse brain genes was upregulated. A GOstat analysis predicted that the upregulated genes were involved in the host immune response. Genes whose expression correlated positively and negatively with parasite numbers were involved in the host immune response, and neuronal morphogenesis and lipid metabolic processes, respectively. These results suggest that changes in the gene expression profile associated with neuronal functions as well as immune responses can contribute to the pathogenesis in N. caninum-infected animals.

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Gene expression analysis by quantitative PCR using RNA from the brain samples used in RNA-seq analysis.(A) Expression of the genes (Saa3, Cxcl9, Ccl8, Ccl5, Cxcl10, Cxcr6, Tgtp2, Gbp8 and Ligp1) that upregulated by N. caminum infection. (B) Expression of the genes (Slc6a4, Slc6a5, Tph2 and Ldlr) that downregulated in symptomatic mice infected with N. caninum. *p < 0.05 and **p < 0.01.
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f4: Gene expression analysis by quantitative PCR using RNA from the brain samples used in RNA-seq analysis.(A) Expression of the genes (Saa3, Cxcl9, Ccl8, Ccl5, Cxcl10, Cxcr6, Tgtp2, Gbp8 and Ligp1) that upregulated by N. caminum infection. (B) Expression of the genes (Slc6a4, Slc6a5, Tph2 and Ldlr) that downregulated in symptomatic mice infected with N. caninum. *p < 0.05 and **p < 0.01.

Mentions: To identify the differentially expressed genes in the experimental groups, we analyzed the transcriptome data from the N. caninum-infected and uninfected mouse brains with DESeq. The magnitude distribution of the significantly altered genes was illustrated with an MA plot analysis (See supplementary Figure S1). We analyzed 37,306 genes including 22,661 protein-coding genes. The expression of 772 protein-coding genes (2.1% of all analyzed genes and 3.4% of protein-coding genes) in the brains of the mice infected with N. caninum was significantly upregulated relative to their expression in the uninfected mice. The 30 most upregulated genes after infection with N. caninum are listed in Table 1. These 30 genes included genes for chemokines and chemokine receptors (Cxcl9, Ccl8, Ccl5, Cxcl10, and Cxcr6), immunoglobulins (Ighg2c, Iglc2, and Igj), interferon (IFN)-inducible GTPase family members (Tgtp2, Gbp8, and Iigp1), and MHC class II antigens (Cd74, H2-Eb1, H2-Aa, and H2-Q7). The quantitative PCR analysis showed that expression of Ccl8, Ccl5, Cxcl10, Cxcr6, Tgtp2, Gbp8, Iigp1 and Saa3 were significantly upregulated in the brains of N. caninum-infected mice (See Figure 4). In contrast, only three genes (Fcrls, Myoc, and Gkn3) were significantly downregulated in the infected mice (See supplementary Table S3). When the functional annotations of the genes upregulated by N. caninum infection were analyzed using GOstat, gene ontology (GO) terms associated with immune system processes, immune responses, and cell activation were represented significantly more strongly in the upregulated genes compared with the reference genes (See Table 2).


Transcriptome and histopathological changes in mouse brain infected with Neospora caninum.

Nishimura M, Tanaka S, Ihara F, Muroi Y, Yamagishi J, Furuoka H, Suzuki Y, Nishikawa Y - Sci Rep (2015)

Gene expression analysis by quantitative PCR using RNA from the brain samples used in RNA-seq analysis.(A) Expression of the genes (Saa3, Cxcl9, Ccl8, Ccl5, Cxcl10, Cxcr6, Tgtp2, Gbp8 and Ligp1) that upregulated by N. caminum infection. (B) Expression of the genes (Slc6a4, Slc6a5, Tph2 and Ldlr) that downregulated in symptomatic mice infected with N. caninum. *p < 0.05 and **p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4300462&req=5

f4: Gene expression analysis by quantitative PCR using RNA from the brain samples used in RNA-seq analysis.(A) Expression of the genes (Saa3, Cxcl9, Ccl8, Ccl5, Cxcl10, Cxcr6, Tgtp2, Gbp8 and Ligp1) that upregulated by N. caminum infection. (B) Expression of the genes (Slc6a4, Slc6a5, Tph2 and Ldlr) that downregulated in symptomatic mice infected with N. caninum. *p < 0.05 and **p < 0.01.
Mentions: To identify the differentially expressed genes in the experimental groups, we analyzed the transcriptome data from the N. caninum-infected and uninfected mouse brains with DESeq. The magnitude distribution of the significantly altered genes was illustrated with an MA plot analysis (See supplementary Figure S1). We analyzed 37,306 genes including 22,661 protein-coding genes. The expression of 772 protein-coding genes (2.1% of all analyzed genes and 3.4% of protein-coding genes) in the brains of the mice infected with N. caninum was significantly upregulated relative to their expression in the uninfected mice. The 30 most upregulated genes after infection with N. caninum are listed in Table 1. These 30 genes included genes for chemokines and chemokine receptors (Cxcl9, Ccl8, Ccl5, Cxcl10, and Cxcr6), immunoglobulins (Ighg2c, Iglc2, and Igj), interferon (IFN)-inducible GTPase family members (Tgtp2, Gbp8, and Iigp1), and MHC class II antigens (Cd74, H2-Eb1, H2-Aa, and H2-Q7). The quantitative PCR analysis showed that expression of Ccl8, Ccl5, Cxcl10, Cxcr6, Tgtp2, Gbp8, Iigp1 and Saa3 were significantly upregulated in the brains of N. caninum-infected mice (See Figure 4). In contrast, only three genes (Fcrls, Myoc, and Gkn3) were significantly downregulated in the infected mice (See supplementary Table S3). When the functional annotations of the genes upregulated by N. caninum infection were analyzed using GOstat, gene ontology (GO) terms associated with immune system processes, immune responses, and cell activation were represented significantly more strongly in the upregulated genes compared with the reference genes (See Table 2).

Bottom Line: A GOstat analysis predicted that the upregulated genes were involved in the host immune response.Genes whose expression correlated positively and negatively with parasite numbers were involved in the host immune response, and neuronal morphogenesis and lipid metabolic processes, respectively.These results suggest that changes in the gene expression profile associated with neuronal functions as well as immune responses can contribute to the pathogenesis in N. caninum-infected animals.

View Article: PubMed Central - PubMed

Affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido, Japan.

ABSTRACT
Neospora caninum is a protozoan parasite that causes neurological disorders in dogs and cattle. It can cause nonsuppurative meningoencephalitis and a variety of neuronal symptoms are observed, particularly in dogs. However, the pathogenic mechanism, including the relationship between the parasite distribution and the clinical signs, is unclear. In this study, to understand the pathogenic mechanism of neosporosis, parasite distribution and lesions were assessed in the brain of mice infected with N. caninum (strain Nc-1). Host gene expression was also analyzed with RNA sequencing (RNA-Seq). The histopathological lesions in the frontal lobe and the medulla oblongata were significantly more severe in symptomatic mice than in asymptomatic mice, although no association between the severity of the lesions and parasite numbers was found. In infected mice, the expression of 772 mouse brain genes was upregulated. A GOstat analysis predicted that the upregulated genes were involved in the host immune response. Genes whose expression correlated positively and negatively with parasite numbers were involved in the host immune response, and neuronal morphogenesis and lipid metabolic processes, respectively. These results suggest that changes in the gene expression profile associated with neuronal functions as well as immune responses can contribute to the pathogenesis in N. caninum-infected animals.

Show MeSH
Related in: MedlinePlus