Sperm-borne small RNAs improve the developmental competence of pre-implantation cloned embryos in rabbit

Qin, Hongyu; Qu, Pengxiang; Hu, Huizhong; Cao, Wenbin; Liu, Hengchao; Zhang, Yanru; Zhao, Jinpeng; Fatima, Nazira; Liu, Enqi

doi:10.1017/s0967199420000805

Cited by 9 publications

(3 citation statements)

References 33 publications

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“…Sperm-borne small RNA injection of cloned bovine embryos enhanced the developmental competence and significantly increased the live birth rate and decreased the birth weights of offspring [152]. Similarly, the injection of sperm small RNA into cloned rabbit embryos increased the total cell number and H3K9me3 level as well as decreased the apoptosis index of cloned rabbit blastocysts [153].…”

Section: The Use Of Non-chemical and Biological Agents As Epigenetic ...mentioning

confidence: 93%

Strategies to Improve the Efficiency of Somatic Cell Nuclear Transfer

Srirattana

Kaneda

Parnpai

2022

IJMS

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Mammalian oocytes can reprogram differentiated somatic cells into a totipotent state through somatic cell nuclear transfer (SCNT), which is known as cloning. Although many mammalian species have been successfully cloned, the majority of cloned embryos failed to develop to term, resulting in the overall cloning efficiency being still low. There are many factors contributing to the cloning success. Aberrant epigenetic reprogramming is a major cause for the developmental failure of cloned embryos and abnormalities in the cloned offspring. Numerous research groups attempted multiple strategies to technically improve each step of the SCNT procedure and rescue abnormal epigenetic reprogramming by modulating DNA methylation and histone modifications, overexpression or repression of embryonic-related genes, etc. Here, we review the recent approaches for technical SCNT improvement and ameliorating epigenetic modifications in donor cells, oocytes, and cloned embryos in order to enhance cloning efficiency.

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Section: The Use Of Non-chemical and Biological Agents As Epigenetic ...mentioning

confidence: 93%

Strategies to Improve the Efficiency of Somatic Cell Nuclear Transfer

Srirattana

Kaneda

Parnpai

2022

IJMS

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“…SCNT was carried out as previously reported [14]. Three drops of micromanipulation M2 medium containing 7.5 μg/ml cytochalasin B and 10% FBS and one drop of 5% PVP (Sigma–Aldrich) were put into a 35 mm dish (Corning, NY, USA) and covered with mineral oil (Sigma–Aldrich).…”

Section: Methodsmentioning

confidence: 99%

Sperm‐borne proteins improve rabbit cloning efficiency via regulating embryonic cleavage and epigenetics

Cao

Zhang

et al. 2022

Proteomics

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Somatic cell nuclear transfer (SCNT) shows great application value in the generation of transgenic animals, protection of endangered species, and therapeutic cloning.However, the cloning efficiency is still very low, which greatly restricts its application. Compared to fertilized embryos, cloned embryos lack the sperm proteins, which are considered to play an important role in embryonic development. Here, we compared the sperm proteome, with that of donor fibroblasts and oocytes, and identified 342 proteins unique to sperm, with 42 being highly expressed. The 384 proteins were mainly enriched in the categories of post-translational modification and cytoskeletal arrangement. Extracts of soluble sperm or fibroblast proteins were injected into cloned embryos, and the result showed that injection of sperm protein significantly inhibited abnormal embryonic cleavage, significantly decreased the level of trimethylated histone H3 Lys9 (H3K9me3) and the apoptotic index, and increased the inner cell mass (ICM)-to-trophectoderm (TE) ratio. More importantly, the sperm proteins also significantly enhanced the birthrate. The results of in vitro and in vivo experiments demonstrate that sperm-derived proteins improve embryo cloning efficiency. Our findings not only provide new insights into ways to overcome low cloning efficiency, but also add to the understanding of sperm protein function.

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“…The role of sperm-borne miR-34c in the first cleavage division of murine embryos remains unclear [ 8 , 20 ], but an increasing number of studies have shown that miR-34c participates in early embryonic development. For example, in rabbits and cows, miR-34c improves the developmental competence of embryos generated by somatic cell nuclear transfer (SCNT) [ 10 , 21 ]. In addition, we have demonstrated that spermatozoan miR-34c levels are correlated with intracytoplasmic sperm injection (ICSI) outcomes, such as embryo quality and clinical pregnancy and live birth rates [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

Sperm-borne microRNA-34c regulates maternal mRNA degradation and preimplantation embryonic development in mice

Cui

Zhuang

et al. 2023

Reprod Biol Endocrinol

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Background Studies have shown that sperm-borne microRNAs (miRNAs) are involved in mammalian preimplantation embryonic development. In humans, spermatozoan miR-34c levels are correlated with in vitro fertilization outcomes, such as embryo quality and the clinical pregnancy and live birth rates. In rabbits and cows, miR-34c improves the developmental competence of embryos generated by somatic cell nuclear transfer. However, the mechanisms underlying the regulation of embryonic development by miR-34c remain unknown. Methods Female C57BL/6 mice (6–8 weeks old) were superovulated, and pronucleated zygotes were collected and microinjected with an miR-34c inhibitor or a negative-control RNA. The embryonic development of the microinjected zygotes was evaluated, and the messenger RNA (mRNA) expression profiles of the embryos at the two-cell, four-cell and blastocyst stages (five embryos per group) were determined by RNA sequencing analysis. Gene expression levels were verified by reverse transcription–quantitative polymerase chain reaction. Cluster analysis and heat map visualization were performed to detect differentially expressed mRNAs. Pathway and process enrichment analyses were performed using ontology resources. Differentially expressed mRNAs were systematically analyzed using the Search Tool for the Retrieval of Interacting Genes/Proteins database to determine their biological functions. Results Embryonic developmental potential was significantly reduced in zygotes microinjected with the miR-34c inhibitor compared with those microinjected with a negative-control RNA. Two-cell stage embryos microinjected with an miR-34c inhibitor presented altered transcriptomic profiles, with upregulated expression of maternal miR-34c target mRNAs and classical maternal mRNAs. Differentially expressed transcripts were mainly of genes associated with lipid metabolism and cellular membrane function at the two-cell stage, with cell-cycle phase transition and energy metabolism at the four-cell stage; and with vesicle organization, lipid biosynthetic process and endomembrane system organization at the blastocyst stage. We also showed that genes related to preimplantation embryonic development, including Alkbh4, Sp1, Mapk14, Sin3a, Sdc1 and Laptm4b, were significantly downregulated after microinjection of an miR-34c inhibitor. Conclusions Sperm-borne miR-34c may regulate preimplantation embryonic development by affecting multiple biological processes, such as maternal mRNA degradation, cellular metabolism, cell proliferation and blastocyst implantation. Our data demonstrate the importance of sperm-derived miRNAs in the development of preimplantation embryos.

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Sperm-borne small RNAs improve the developmental competence of pre-implantation cloned embryos in rabbit

Cited by 9 publications

References 33 publications

Strategies to Improve the Efficiency of Somatic Cell Nuclear Transfer

Strategies to Improve the Efficiency of Somatic Cell Nuclear Transfer

Sperm‐borne proteins improve rabbit cloning efficiency via regulating embryonic cleavage and epigenetics

Sperm-borne microRNA-34c regulates maternal mRNA degradation and preimplantation embryonic development in mice

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