Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Nov 17:13:1053826.
doi: 10.3389/fgene.2022.1053826. eCollection 2022.

RNA interference analysis of potential functions of cyclin A in the reproductive development of male oriental river prawns (Macrobrachium nipponense)

Wenyi Zhang  1 , Yiwei Xiong  1 , Pengchao Wang  2 , Tianyong Chen  3 , Sufei Jiang  1 , Hui Qiao  1 , Yongsheng Gong  1 , Yan Wu  1 , Shubo Jin  1   2 , Hongtuo Fu  1   2
Affiliations

RNA interference analysis of potential functions of cyclin A in the reproductive development of male oriental river prawns (Macrobrachium nipponense)

Wenyi Zhang et al. Front Genet. .

Abstract

Cyclin A (CycA) plays essential roles in regulating multiple steps of the cell cycle, and it affects gonad development in mammals and invertebrates. Previous RNA interference (RNAi) analysis revealed that knocking-down the expression of CycA in female oriental river prawns (Macrobrachium nipponense) inhibited ovarian development. CycA was also predicted to have regulatory roles in reproductive development of male M. nipponense based on significant changes of Mn-CycA expression after eyestalk ablation. The goal of this study was to investigate the potential functions of CycA in the reproductive development of male M. nipponense using RNAi and histological observations. Quantitative real-time PCR analysis revealed that both single-side and double-side eyestalk ablation stimulated the expressions of Mn-CycA, and the expression was higher in prawns with double-side eyestalk ablation (p < 0.05). Mn-CycA expression was significantly higher in the testis and androgenic gland during the reproductive season than during the non-reproductive season (p < 0.05). In the RNAi analysis, Mn-CycA expression significantly decreased after prawns were injected with dsCycA, and the expression of insulin-like androgenic gland hormone (Mn-IAG) also decreased as Mn-CycA expression decreased. This result indicated that CycA positively regulated the expression of IAG in M. nipponense. Histological observations revealed that the number of sperm decreased dramatically to <5% of the total cells in the testis of the dsCycA-treated group compared to that of control group on day 14, indicating that knockdown of Mn-CycA expression inhibited testis development by affecting the expression of Mn-IAG in M. nipponense. These results highlighted the functions of CycA in male reproductive development of M. nipponense, which can be applied to future studies of male reproduction in other crustacean species.

Keywords: Macrobrachium nipponense; RNA interference; cyclin A; insulin-like androgenic gland hormone; male reproduction.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Measurement of the Mn-CycA expressions after the ablations of single-side and double-side eyestalk from male M. nipponense by qPCR. The amount of Mn-CycA mRNA was normalized to the EIF transcript level. Data are shown as mean ± SD (standard deviation) of tissues from three separate individuals. Lowercases indicate expression difference between different days in the same group (p < 0.05). Capital letters indicated the expression difference between different groups at the same day (p < 0.05).
FIGURE 2
FIGURE 2
Measurement of the Mn-CycA expression in the testis and androgenic gland taken from different reproductive season. The amount of Mn-CycA mRNA was normalized to the EIF transcript level. Data are shown as mean ± SD (standard deviation) of tissues from three separate individuals. Lowercases indicate expression difference between different samples. (A) Mn-CycA expression in the testes taken from different reproductive season. (B) Mn-CycA expression in the androgenic gland taken from different reproductive season.
FIGURE 3
FIGURE 3
In situ hybridization analysis of Mn-CycA in the testis and androgenic gland taken from the reproductive season. SG: Spermatogonia; SC: spermatocyte; S: sperm; AGC: androgenic gland cells; EB: ejaculatory bulb. Scale bars = 20 μm.
FIGURE 4
FIGURE 4
Measurement of Mn-CycA and Mn-IAG expression at different days after dsCycA and dsGFP injection. The amount of Mn-CycA and Mn-IAG mRNA was normalized to the EIF transcript level. Data are shown as mean ± SD (standard deviation) of tissues from three separate individuals. Lowercases indicated expression difference between different days after dsGFP and dsCycA injection. ** (p < 0.01) indicates significant expression difference between the dsCycA and dsGFP treated prawns at the sample day. (A) Measurement of Mn-CycA expression at different days after dsGFP and dsCycA injection. (B) Measurement of Mn-IAG expression at different days after dsGFP and dsCycA injection.
FIGURE 5
FIGURE 5
The histological observations of testis between dsCycA and dsGFP treated prawns. SG: Spermatogonia; SC: spermatocyte; S: sperm. Scale bars = 20 μm.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Almeida E. A., Petersen R. L., Andreatta E. R., Bainy A. C. (2004). Effects of captivity and eyestalk ablation on antioxidant status of shrimps (Farfantepenaeus paulensis). Aquaculture 238, 523–528. 10.1016/j.aquaculture.2004.04.010 - DOI
    1. Bergman Z. J., McLaurin J. D., Eritano A. S., Johnson B. M., Sims A. Q., Riggs B. (2015). Spatial reorganization of the endoplasmic reticulum during mitosis relies on mitotic kinase cyclin A in the early Drosophila embryo. PLoS ONE 10, e0117859. 10.1371/journal.pone.0117859 - DOI - PMC - PubMed
    1. Bourouh M., Dhaliwal R., Rana K., Sinha S., Guo Z., Swan A. (2016). Distinct and overlapping requirements for cyclins A, B, and B3 in drosophila female meiosis. G3 6, 3711–3724. 10.1534/g3.116.033050 - DOI - PMC - PubMed
    1. Diarte-Plata G., Sainz-Hernández J. C., Aguiñaga-Cruz J. A., Fierro-Coronado J. A., Polanco-Torres A., Puente-Palazuelos C. (2012). Eyestalk ablation procedures tominimize pain in the freshwater prawn Macrobrachium Americanum. Appl. Anim. Behav. Sci. 140, 172–178. 10.1016/j.applanim.2012.06.002 - DOI
    1. Hopkins P. M. (2012). The eyes have it: A brief history of crustacean neuroendocrinology. Gen. Comp. Endocrinol. 175, 357–366. 10.1016/j.ygcen.2011.12.002 - DOI - PubMed

LinkOut - more resources