Characteristics of SP600125 Induced Tetraploid Cells in Comparison With Diploid and Tetraploid Cells of Fish.

Yunpeng Fan,Li Ren,Yamei Xiao,Wenbin Liu,Shujuan Chen,Shaojun Liu,Liangyue Peng,Guangjing Zhang,Kaiyue Zhao,Jinhui Liu,Wen Fu

doi:10.3389/fgene.2021.781007

Abstract

In our previous research, SP600125 (Anthrapyrazolone) was used to induce autotetraploid of crucian carp cells (SP4N cells), and tetraploid fry was generated from the SP4N cells by somatic cell nuclear transfer technique. However, it is still unclear about biological characteristics of the SP4N cells. In this article, the cytological characteristic and gene expression profiles of the SP4N cells are investigated in comparison with the crucian carp cells (2N cells) and the tetraploid crucian carp cells (CC4N cells). The SP4N cells have tetraploid characteristics in terms of morphology and DNA ploidy levels, and their chromosome behavior is stable during the cell proliferation. The migration ability and the mtDNA copy number of SP4N cells are both lower than those in the CC4N cells and the 2N cells, but there exist giant mitochondria in the SP4N cells. The similar expression trends in the cell cycle regulation genes of the SP4N cells and 2N cells, while the corresponding expression profiles are clearly different between the SP4N cells and the CC4N cells. Moreover, the significant difference genes are associated with energy metabolism pathways among the SP4N cells, 2N cells and CC4N cells. These results can provide deeper understanding of SP600125 induction, as well as finding applications in polyploidization breeding of fish species.

Highlights

Small compound SP600125 (Anthrapyrazolone), known as a c-Jun N-terminal kinase (JNK) inhibitor, is often used in research on JNK signaling pathway (Bennett et al, 2001)
Flow cytometry analysis further demonstrated that there existed diploid (2x, 100) and tetraploid (4x, 200) peaks in the 2N cells, while the tetraploid (4x, 200) and octoploid (8x, 400) peaks display in the SP4N and CC4N cells (Figure 1C)
We proved that the SP4N cells had a normal chromosome behavior in cell proliferation

Summary

Introduction

Small compound SP600125 (Anthrapyrazolone), known as a c-Jun N-terminal kinase (JNK) inhibitor, is often used in research on JNK signaling pathway (Bennett et al, 2001). Our previous study found that SP600125 could induce polyploidization in diploid crucian carp cells, and generated an autotetraploid cell line (named SP4N cells) (Zhou et al, 2016). Combining somatic cell nuclear transfer technique, autotetraploid fish was successfully generated from the SP4N cells (Zhou et al, 2016). Little is known about the biological characteristics of the SP4N cells. Polyploid is rare phenomenon in animals (Comai 2005; Blomme et al, 2006; Schoenfelder and Fox 2015). Tetraploid crucian carp is hybrids of Carassius auratus red var. Cytological and transcriptome analysis will be conducted for the SP4N cells, the diploid cells from the caudal fin of C. auratus (2N cells), and the tetraploid cells from the caudal fin of tetraploid crucian carp (CC4N cells). We attempt to answer the following question: Compared with the 2N cells and the CC4N

Methods

Results

Conclusion