Identification of circular RNAs in the immature and mature rat anterior pituitary

in Journal of Endocrinology
Correspondence should be addressed to B Yuan or J-B Zhang: yuan_bao@jlu.edu.cn or zjb515@126.com
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Circular RNAs (circRNAs) are a new class of RNA that have a stable structure characterized by covalently closed circular molecules and are involved in invasive pituitary adenomas and recurrent clinically nonfunctioning pituitary adenomas. However, information on circRNAs in the normal pituitary, especially in rats, is limited. In this study, we identified 4123 circRNAs in the immature (D15) and mature (D120) rat anterior pituitary using the Illumina platform, and 32 differentially expressed circRNAs were found. A total of 150 Gene Ontology terms were significantly enriched, and 16 KEGG pathways were found to contain differentially expressed genes. Moreover, we randomly selected eight highly expressed circRNAs and detected their relative expression levels in the mature and immature rat pituitary by qPCR. In addition, we predicted 90 interactions between 53 circRNAs and 57 miRNAs using miRanda. Notably, circ_0000964 and circ_0001303 are potential miRNA sponges that may regulate the Fshb gene. The expression profile of circRNAs in the immature and mature rat anterior pituitary may provide more information about the roles of circRNAs in the development and reproduction in mammals.

Downloadable materials

  • Table S1. Primers used for qPCR
  • Table S2.The chromosomes distribution of circRNAs.
  • Table S3. 32 differentially expressed circRNAs.
  • Table S4. 150 significantly enriched GO terms.
  • Table S5. 16 KEGG pathways contains with differentially expressed genes.
  • Table S6. interation between circRNAs and miRNAs

 

      Society for Endocrinology

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    Deep sequencing of circRNAs in rats. (A) Identification pipeline for circRNAs. (B) Classification of 4123 circRNAs, including classic, alter exon, intron, overlap exon, antisense and intergenic circRNAs. (C) Distribution of circRNAs on the rat chromosomes. A full color version of this figure is available at https://doi.org/10.1530/JOE-18-0540.

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    Expression pattern of circRNAs in the mature and immature rat anterior pituitary. Distributions of the sequence length (A), exon number (B) and exon length (C) of the circRNAs. (D) Boxplots of the expression levels (SRPBM value) of circRNAs in D15 and D120 rats. A full color version of this figure is available at https://doi.org/10.1530/JOE-18-0540.

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    Identification of differentially expressed circRNAs. (A) Proportion of upregulated and downregulated circRNAs in the rat anterior pituitary at different developmental stages. (B) Volcano plot analysis of differentially expressed circRNAs between D15 and D120. Red dots indicate upregulated genes; green dots represent downregulated genes. (C) Analysis of the expression patterns of differentially expressed circRNAs. The highest to lowest fold changes are marked from red to blue, respectively. A full color version of this figure is available at https://doi.org/10.1530/JOE-18-0540.

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    Enrichment of differentially expressed circRNAs. The GO annotation of upregulated and downregulated mRNAs covers the fields of cellular components, biological processes and molecular functions. Red columns indicate upregulated genes, and green columns indicate downregulated genes. A full color version of this figure is available at https://doi.org/10.1530/JOE-18-0540.

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    Validation of highly expressed circRNAs and two key circRNAs. (A) Expression of upregulated and downregulated circRNAs in the mature and immature rat anterior pituitary. (B and C) Expression levels of circ_0000964 and circ_0001303 in the mature and immature rat anterior pituitary. All experiments were repeated at least three times. The data are shown as the mean ± s.d. Statistical significance was analyzed by one-way ANOVA, P < 0.05 was considered significant, and differences are marked with the letters a and b. A full color version of this figure is available at https://doi.org/10.1530/JOE-18-0540.

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    Interactions between circRNAs and miRNAs. The network was based on the miRanda program. The triangles represent the circRNAs, and the V shape represents the miRNAs. The pink color indicates the components that are upregulated in the mature vs immature rat pituitary, the purple color indicates the components that are downregulated, and the gray color indicates the components with no change in either group. The size of the graphs represents the expression of miRNAs and circRNAs, and a larger graph indicates higher expression. A full color version of this figure is available at https://doi.org/10.1530/JOE-18-0540.

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