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. 2019 Dec;21(6):753-761.
doi: 10.1007/s10126-019-09921-x. Epub 2019 Sep 14.

Analysis on the Meiosis-Related Gene (Dmc1, Ph1) Expression in Autotriploid Carassius auratus

Qinbo Qin  1 , Yuwei Zhou  1 , Chongqing Wang  1 , Minghe Zhang  1 , Huan Qin  1 , Chun Zhao  1 , Shaojun Liu  2
Affiliations

Analysis on the Meiosis-Related Gene (Dmc1, Ph1) Expression in Autotriploid Carassius auratus

Qinbo Qin et al. Mar Biotechnol (NY). 2019 Dec.

Abstract

Triploid is usually considered to be unable to perform normal meiosis due to the abnormal behavior of the three sets of chromosomes. But autotriploid Carassius auratus in the Dongting water system (3n = 150, abbreviated as 3nCC) can perform normal meiosis. In artificial autotriploid Carassius auratus (3n = 150, abbreviated as 3nRR), female individuals undergo normal meiosis and produce mature gametes, while male individuals cannot. To better understand the effects of triploidization on meiosis in fish, we study the structure, methylation level, and expression level of meiosis-related genes (Dmc1, Ph1) in diploid Carassius auratus (2n = 100, abbreviated as 2nCC), Carassius auratus red var.(2n = 100, abbreviated as RCC), 3nCC and 3nRR. The results show that, compared with their diploid ancestors (2nCC and RCC), Dmc1 and Ph1 genes are hypomethylated in all 3nCC and female 3nRR, while are hypermethylated in male 3nRR. Correspondingly, Dmc1 and Ph1 genes are highly expressed in all 3nCC and female 3nRR, while are lowly expressed in male 3nRR. These results indicate that high expression of meiosis-related genes can contribute to restoration of bivalent pairing during meiosis in autotriploid Carassius auratus. This study provides new insights into the effect of DNA methylation on the fertility in triploid fish.

Keywords: Expression; Meiosis; Meiosis-related gene; Methylation; Triploid.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Formation of 3nCC. (A) The formation of 3nCC. (B) The mean DNA content of 3nCC. (C) The mean DNA content of 3nCC sperm. 3nCC, autotriploid Carassius auratus
Fig. 2
Fig. 2
Formation of 3nRR. (A) The formation of 3nCC. (B), (C) Fish hybridisation signals in the metaphase chromosomes of RCC and 3nRR with class III (477 bp) 5S rDNA as a probe. The white arrows indicate the 5S rDNA gene loci. The eight 5S gene loci in RCC (A) and twelve in 3nRR (B). The bars in (A ,B) denote 3 μm. RCC, Carassius auratus red var.; 3nRR, autotriploid
Fig. 3
Fig. 3
Gonad microstructure of the reproductive stage. (A) RCC ovary; (B) RCC testis; (C) 3nRR ovary; (D) 3nRR testis; (E) 2nCC ovary; (F) 2nCC testis; (G) 3nCC ovary; (H) 3nCC testis. In the picture, the red arrow points to the spermatocyte, and the black arrow points to the sperms. As the figure shows, 3nCC and 3nRR containing many phase III oocytes. The 3nCC testis contains a large number of mature sperm, while the sperm in the 3nRR testis are vacuolated or broken. Bar: 50 μm
Fig. 4
Fig. 4
Relative expression of the Ph1 and Dmc1 genes in the gonads of RCC, 3nRR, 2nCC and 3nCC during the breeding season. Among them, (A) and (C) are the relative expression levels of Ph1 and Dmc1 in the ovary. (B) and (D) are the relative expression levels of Ph1 and Dmc1 in the testis
Fig. 5
Fig. 5
Sequencing results of methylation extent of the promoter regions of Ph1 and Dmc1 genes, wherein yellow represents methylation and blue represents no methylation
Fig. 6
Fig. 6
Correlation map of methylation level and expression level
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