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Vertebrate reproductive science and technology
Reproduction, Fertility and Development

Reproduction, Fertility and Development

Volume 31 Number 2 2019

RD18178Single-cell RNA sequencing reveals distinct gene expression patterns in glucose metabolism of human preimplantation embryos

Di-Cheng Zhao, Yu-Mei Li, Jie-Liang Ma, Ning Yi, Zhong-Yuan Yao, Yan-Ping Li, Yi Quan, Xin-Ning Li, Chang-Long Xu, Ying Qiu and Ling-Qian Wu 0000-0002-7070-5896
pp. 237-247

Precise regulation of glucose metabolism-related genes is essential for early embryonic development. For the first time, we did integrated analysis of single-cell RNA-sequencing (scRNA-seq) data of human pre-implantation embryos. We discovered that the gene expression profiles are dynamic according to glucose utilization in the embryos at different stages.

RD18034Comparative analysis of testis transcriptomes associated with male infertility in triploid cyprinid fish

Wuhui Li, Hui Tan, Junmei Liu, Jie Hu, Jialin Cui, Shi Wang, Qingfeng Liu, Fangzhou Hu, Li Ren, Min Tao, Rurong Zhao, Conghui Yang, Qinbo Qin and Shaojun Liu 0000-0002-1960-2533
pp. 248-260

Continuous histological observation and testis transcriptomes were combined to investigate the fertility of male triploid fish. Differential expression of pathways including Wnt signalling, apoptosis and ubiquitin-mediated proteolysis, as well as a number of genes associated with spermatogenesis, was identified in the testis of triploid fish, which may result in infertility. This study provides information regarding the potential molecular regulatory mechanisms underlying male infertility in polyploid fish.


Because Andrias davidianus is an endangered species, it is necessary to investigate the regulation of development and gonad differentiation in this species. This study revealed the molecular characteristics of oestrogen receptor ERα in A. davidianus. Exposure of larvae to bisphenol A (BPA) led to a decrease in ERα expression in the testis, but had no significant effect on the sex ratio during sexual differentiation. The effects of BPA on the life cycle of A. davidianus and the potential mechanism underlying BPA resistance in A. davidianus will be further investigated in future studies.

RD17527Astaxanthin improves the developmental competence of in vitro-grown oocytes and modifies the steroidogenesis of granulosa cells derived from bovine early antral follicles

M. A. Abdel-Ghani, Y. Yanagawa, A. Z. Balboula, K. Sakaguchi, C. Kanno, S. Katagiri, M. Takahashi and M. Nagano 0000-0002-1890-6842
pp. 272-281

In vitro growth (IVG) culture is not established for bovine oocytes and development of the culture system is necessary. This study investigated the effect of astaxanthin (Ax), a strong antioxidant, on the developmental competence of bovine oocytes during IVG. Ax supplementation of IVG medium improved the quality of bovine oocytes due to suppression of the luteinisation of granulosa cells.


Polycystic ovary syndrome (PCOS) is a common cause of infertility due to anovulation and abnormal folliculogenesis. This study investigated the effect of l-carnitine on folliculogenesis in mice following induction of PCOS. l-Carnitine improved the function and structure of the ovary through desired changes in folliculogenesis. l-Carnitine is therefore suggested as a therapeutic supplement in the case of PCOS.


Artificial blastocoel collapse is a technique commonly used in IVF to improve embryo viability after vitrification and warming, but the mechanisms by which it enhances survival are unknown. We found an increase in active fluid transport in artificially collapsed embryos. In addition, there were no changes in markers of embryonic stress as a result of this technique, providing further support for its safety in an IVF setting.

RD17446In vitro culture and non-invasive metabolic profiling of single bovine embryos

Monika Nõmm 0000-0002-3627-2126, Rando Porosk, Pille Pärn, Kalle Kilk, Ursel Soomets, Sulev Kõks and Ülle Jaakma
pp. 306-314

Selecting high-quality embryos for transfer has been a difficult task when producing bovine embryos in vitro. The aims of this study were to develop a non-invasive culture medium sample collection method and to determine potential viability markers for embryo development in bovine in vitro production. Metabolic profiling provides a tool to identify well-developing embryos in a non-invasive manner before transfer.


Insulin resistance (IR) and infertility are two major complications of polycystic ovary syndrome (PCOS). We found that quercetin as a natural flavonoid improved the IR state and increased expression of two important fertility related genes in the uterus of PCOS rats; it could thus be a possible treatment for PCOS and infertility.

RD18203Extracellular vesicles derived from donor oviduct fluid improved birth rates after embryo transfer in mice

Pengxiang Qu, Yuelei Zhao, Rong Wang, Yali Zhang, Lu Li, Jianglin Fan and Enqi Liu
pp. 324-332

The lower success rate of embryo transfer in mice constitutes a major problem in its applications. We demonstrated here that extracellular vesicles derived from oviduct fluid significantly improved efficiency of mouse embryo transfer through inhibiting cell apoptosis and enhancing cell differentiation. Therefore, extracellular vesicles may become a novel factor to increase the success rate of mouse embryo transfer in future.


Conventional media supplemented with fetal bovine serum (FBS) have the disadvantage of variations in serum from lot-to-lot that can affect the reproducibility of experimental findings. Here, we showed that supplementation of in vitro culture medium with insulin–transferrin–sodium selenite plus epidermal growth factor and bovine serum albumin, as alternatives to FBS, can consistently support the development of in vitro-produced bovine embryos and effectively improve invasion ability of trophoblasts via induction of matrix metalloproteinases.


Maternal factors Dppa3 and Tet3 in oocytes regulate DNA demethylation in fertilised eggs; histone H3K9 dimethylation (me2) is critical in this event. This study shows that contrary to the situation in fertilised eggs, H3K9me2 is not functionally associated with Dppa3 and Tet3 in somatic cells or somatic cell cloned embryos. These results provide new insights for understanding the function of Dppa3 and the mechanism of DNA demethylation in cells and embryos.

RD17357Zebularine significantly improves the preimplantation development of ovine somatic cell nuclear transfer embryos

Hui Cao, Jun Li 0000-0002-8475-8040, Wenlong Su, Junjie Li, Zhigang Wang, Shuchun Sun, Shujun Tian, Lu Li, Hanyang Wang, Jiexin Li, Xiaohuan Fang, Qiaoli Wei and Chuang Liu
pp. 357-365

Zebularine improves the efficiency of somatic cell nuclear transfer. This study used zebularine in ovine cloning and found that zebularine decreased DNA methylation levels overall and in the promoters of pluripotency genes in donor cells, leading to improved pluripotency gene expression and ultimately increasing the developmental potential of cloned embryos. Thus, zebularine is beneficial for ovine cloning.

RD18070Fetal bovine serum promotes the development of in vitro porcine blastocysts by activating the Rho-associated kinase signalling pathway

Shimeng Guo, Shichao Liu, Gerelchimeg Bou, Jia Guo, Liyuan Jiang, Zhuang Chai, Mingming Cai, Yanshuang Mu and Zhonghua Liu 0000-0002-4126-9060
pp. 366-376

In this study we demonstrate the beneficial effects of fetal bovine serum (FBS) in porcine embryo development in vitro. The results may improve our understanding of the mechanism why FPB benefits blastocyst development and help us identify the major beneficial component(s) in FBS or to improve the in vitro culture medium. FBS promoted the in vitro development of embryos by activating Rho-associated kinase signalling in a chemically defined medium.


HOX transcript antisense RNA (HOTAIR) levels in the placenta decreased significantly decreased with increasing gestational age, and were negatively correlated with vascular endothelial growth factor (VEGF) A levels. HOTAIR suppressed the proliferation, migration, invasion and tube formation of human umbilical vein endothelial cells, and inhibited VEGFA transcription by directly targeting the VEGFA promoter. Thus, HOTAIR may represent a potential therapeutic target for patients with human placental vascularisation abnormalities.

RD18102Maturation of buffalo oocytes in vitro with acetyl-L-carnitine improves cryotolerance due to changes in mitochondrial function and the membrane lipid profile

Hui-Yan Xu 0000-0002-1130-5817, Shuang-Shuang Geng, Ting-Ting Li, Qiang Fu, Sheng-Sheng Lu, Xing-Wei Liang, Yang-Qing Lu, Ming Zhang, Xiao-Gan Yang and Ke-Huan Lu
pp. 386-394

This is the first report that acetyl-l-carnitine (ALC) supplementation during IVM affects phospholipids profiles of buffalo oocytes after vitrification. Supplementation with ALC has beneficial effects on mitochondrial function and alters the phospholipid composition of vitrified oocyte membranes. ALC improves the quality of mature buffalo oocytes and increases the success of embryo development after vitrification.

RD17452Sperm functional attributes and oviduct explant binding capacity differs between bulls with different fertility ratings in the water buffalo (Bubalus bubalis)

Kaustubh Kishor Saraf, Raushan Kumar Singh, Arumugam Kumaresan, Samiksha Nayak, Shivani Chhillar, Sreela Lathika, Tirtha Kumar Datta and Tushar Kumar Mohanty
pp. 395-403

Prediction of bull fertility is of major economic value to the cattle breeding industry. We showed that functional attributes and oviduct binding ability of spermatozoa differed between bulls with different field fertility ratings. These findings may help in developing bull fertility prediction tools, once validated on a greater number of bulls.

RD18093Aggregation recovers developmental plasticity in mouse polyploid embryos

Hiroyuki Imai, Wataru Fujii, Ken Takeshi Kusakabe, Yasuo Kiso and Kiyoshi Kano 0000-0002-8967-9536
pp. 404-411

Repeated whole genome duplication (WGD) has been utilised for evolution; however, tolerance for repeated WGD still remains unknown in present-day mammals. A hexadecaploid embryo, produced by repeated electrofusion, developed to blastocyst stage without the inner cell mass in mice. Aggregation of four hexadecaploid embryos recovered the inner cell mass, suggesting the tolerance and higher plasticity to adapt to repeated WGD for blastocyst formation.


This study revealed that cytoplasmic polyadenylation element binding protein 2 (CPEB2) is localised to the apical region of cells from the morula stage onwards and the depletion of CPEB2 disrupts tight-junction (TJ) assembly during porcine preimplantation development. Interestingly, CPEB2 knockdown does not affect the transcript level of TJ-associated genes but alters protein expression and subcellular localisation. Therefore, CPEB2 may mediate translocation and stability of mRNA for genes associated with TJ.

RD18087Melatonin reduces cAMP-stimulated capacitation of ram spermatozoa

Silvia Gimeno-Martos 0000-0002-0455-3409, Adriana Casao, Marc Yeste 0000-0002-2209-340X, José A. Cebrián-Pérez, Teresa Muiño-Blanco and Rosaura Pérez-Pé
pp. 420-431

Melatonin is able to regulate sperm capacitation showing different effects depending on concentration, but its effect on spermatozoa incubated in high cAMP conditions is unknown. Our results indicate that melatonin at micromolar concentrations acts reducing ROS and cAMP levels, whereas at lower concentrations modifies motile sperm subpopulations that change during capacitation. These findings warrant further studies on the potential use of melatonin for controlling capacitation in artificial insemination procedures.

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