New Insights into the Mechanism by Which the Pituitary Gland Copes with Hypoxia Stress Based on a Transcriptomic Analysis of Megalobrama amblycephala

Abstract

Hypoxia is a common environmental stressor in aquatic ecosystems, and during the cultivation process, Megalobrama amblycephala is prone to death because it is hypoxia-intolerant, which brings huge economic losses to farmers. The pituitary gland is a crucial endocrine gland in fish, and it is mainly involved in the secretion, storage, and regulation of hormones. In the present study, we compared the transcriptional responses to serious hypoxia in the pituitary gland among hypoxia-sensitive (HS) and hypoxia-tolerant (HT) M. amblycephala and a control group that received a normal oxygen supply (C0). The fish were categorized according to the time required to lose balance during a hypoxia treatment. A total of 129,251,170 raw reads were obtained. After raw sequence filtering, 43,461,745, 42,609,567, and 42,730,282 clean reads were obtained for the HS, HT, and C0 groups, respectively. A transcriptomic comparison revealed 1234 genes that were differentially expressed in C0 vs. HS, while 1646 differentially expressed genes were obtained for C0 vs. HT. In addition, the results for HS vs. HT showed that 367 upregulated and 41 downregulated differentially expressed genes were obtained for a total of 408 differentially expressed genes. A KEGG analysis of C0 vs. HS, C0 vs. HT, and HS vs. HT identified 315, 322, and 219 enriched pathways, respectively. Similar hypoxia-induced transcription patterns suggested that the downregulated DEGs and enriched pathways were related to pathways of neurodegeneration in multiple diseases, pathways in cancer, thermogenesis, microRNAs in cancer, diabetic cardiomyopathy, and renin secretion. However, in the upregulated DEGs, the PI3K-Akt signaling pathway (C0 vs. HS), microRNAs in cancer (C0 vs. HT), and HIF-1 signaling pathway (HS vs. HT) were significantly enriched. There is a lack of clarity regarding the role of the pituitary gland in hypoxic stress. These results not only provide new insights into the mechanism by which pituitary tissue copes with hypoxia stress in M. amblycephala but also offer a basis for breeding M. amblycephala with hypoxia-resistant traits.

Keywords: Megalobrama amblycephala; hypoxia; pituitary; transcriptomic.

Figure 1

Pictures collected during the experiment. (a) M. amblycephala (9–11 cm and 18–20 g). (b) The symptoms of hypoxia in M. amblycephala. (c) A pituitary gland obtained from M. amblycephala.

Figure 2

The hypoxia device used in this experiment.

Figure 3

The numbers of differentially expressed genes in the three groups of M. amblycephala. The x-axis indicates the groups of M. amblycephala, and the y-axis indicates the numbers of differentially expressed genes.

Figure 4

GO classification of the differentially expressed genes in the three groups of M. amblycephala. (a) C0 vs. HS. (b) C0 vs. HT. (c) HS vs. HT.

Figure 4

GO classification of the differentially expressed genes in the three groups of M. amblycephala. (a) C0 vs. HS. (b) C0 vs. HT. (c) HS vs. HT.

Figure 5

KEGG enrichment of the differentially expressed genes in the three groups of M. amblycephala. (a) C0 vs. HS. (b) C0 vs. HT. (c) HS vs. HT.

Figure 5

KEGG enrichment of the differentially expressed genes in the three groups of M. amblycephala. (a) C0 vs. HS. (b) C0 vs. HT. (c) HS vs. HT.

Figure 6

Validation of RNA-seq data using quantitative real-time polymerase chain reaction (RT-PCR).