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7. Postcopulatory Reproductive Strategies in Spermatozoa

Author

Matsuzaki, Mei, Sasanami, Tomohiro, Iwata, Yoko, Hirohashi, Noritaka, Asami, Takahiro, Series Editor, Kajihara, Hiroshi, Series Editor, Kobayashi, Kazuya, Series Editor, Koizumi, Osamu, Series Editor, Motokawa, Masaharu, Series Editor, Naruse, Kiyoshi, Series Editor, Satoh, Akiko, Series Editor, Takamune, Kazufumi, Series Editor, Takeuchi, Hideaki, Series Editor, Yoshikuni, Michiyasu, Series Editor, Kitano, Takeshi, editor, Iwao, Yasuhiro, editor, and Kondo, Mariko, editor

Published
2018
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27. Cyclin D1 gene expression is essential for cell cycle progression from the maternal-to-zygotic transition during blastoderm development in Japanese quail

Author

1000020515382, Mizushima, Shusei, Sasanami, Tomohiro, Ono, Tamao, Matsuzaki, Mei, Kansaku, Norio, Kuroiwa, Asato, 1000020515382, Mizushima, Shusei, Sasanami, Tomohiro, Ono, Tamao, Matsuzaki, Mei, Kansaku, Norio, and Kuroiwa, Asato

Abstract

Embryogenesis proceeds by a highly regulated series of events. In animals, maternal factors that accumulate in the egg cytoplasm control cell cycle progression at the initial stage of cleavage. However, cell cycle regulation is switched to a system governed by the activated nuclear genome at a specific stage of development, referred to as maternal-to-zygotic transition (MZT). Detailed molecular analyses have been performed on maternal factors and activated zygotic genes in MZT in mammals, fishes and chicken; however, the underlying mechanisms remain unclear in quail. In the present study, we demonstrated that MZT occurred at blastoderm stage V in the Japanese quail using novel gene targeting technology in which the CRISPR/Cas9 and intracytoplasmic sperm injection (ICSI) systems were combined. At blastoderm stage V, we found that maternal retinoblastoma 1 (RB1) protein expression was down-regulated, whereas the gene expression of cyclin D1 (CCND1) was initiated. When a microinjection of sgRNA containing CCND1-targeted sequencing and Cas9 mRNA was administered at the pronuclear stage, blastoderm development stopped at stage V and the down-regulation of RB1 did not occur. This result indicates the most notable difference from mammals in which CCND-knockout embryos are capable of developing beyond MZT. We also showed that CCND1 induced the phosphorylation of the serine/threonine residues of the RB1 protein, which resulted in the degradation of this protein. These results suggest that CCND1 is one of the key factors for RB1 protein degradation at MZT, and the elimination of RB1 may contribute to cell cycle progression after MZT during blastoderm development in the Japanese quail. Our novel technology, which combined the CRISPR/Cas9 system and ICSI, has the potential to become a powerful tool for avian-targeted mutagenesis.

Published
2021

29. Cyclin D1 gene expression is essential for cell cycle progression from the maternal-to-zygotic transition during blastoderm development in Japanese quail

Author

Mizushima, Shusei, Sasanami, Tomohiro, Ono, Tamao, Matsuzaki, Mei, Kansaku, Norio, Kuroiwa, Asato, Mizushima, Shusei, Sasanami, Tomohiro, Ono, Tamao, Matsuzaki, Mei, Kansaku, Norio, and Kuroiwa, Asato

Abstract

Embryogenesis proceeds by a highly regulated series of events. In animals, maternal factors that accumulate in the egg cytoplasm control cell cycle progression at the initial stage of cleavage. However, cell cycle regulation is switched to a system governed by the activated nuclear genome at a specific stage of development, referred to as maternal-to-zygotic transition (MZT). Detailed molecular analyses have been performed on maternal factors and activated zygotic genes in MZT in mammals, fishes and chicken; however, the underlying mechanisms remain unclear in quail. In the present study, we demonstrated that MZT occurred at blastoderm stage V in the Japanese quail using novel gene targeting technology in which the CRISPR/Cas9 and intracytoplasmic sperm injection (ICSI) systems were combined. At blastoderm stage V, we found that maternal retinoblastoma 1 (RB1) protein expression was down-regulated, whereas the gene expression of cyclin D1 (CCND1) was initiated. When a microinjection of sgRNA containing CCND1-targeted sequencing and Cas9 mRNA was administered at the pronuclear stage, blastoderm development stopped at stage V and the down-regulation of RB1 did not occur. This result indicates the most notable difference from mammals in which CCND-knockout embryos are capable of developing beyond MZT. We also showed that CCND1 induced the phosphorylation of the serine/threonine residues of the RB1 protein, which resulted in the degradation of this protein. These results suggest that CCND1 is one of the key factors for RB1 protein degradation at MZT, and the elimination of RB1 may contribute to cell cycle progression after MZT during blastoderm development in the Japanese quail. Our novel technology, which combined the CRISPR/Cas9 system and ICSI, has the potential to become a powerful tool for avian-targeted mutagenesis.

Published
2021

38. A unique mechanism of successful fertilization in a domestic bird

Author

Sasanami Tomohiro, Izumi Shunsuke, Sakurai Naoki, Hirata Toshifumi, Mizushima Shusei, Matsuzaki Mei, Hiyama Gen, Yorinaga Eriko, Yoshimura Takashi, Ukena Kazuyoshi, Tsutsui Kazuyoshi, Sasanami Tomohiro, Izumi Shunsuke, Sakurai Naoki, Hirata Toshifumi, Mizushima Shusei, Matsuzaki Mei, Hiyama Gen, Yorinaga Eriko, Yoshimura Takashi, Ukena Kazuyoshi, and Tsutsui Kazuyoshi

Abstract

publisher

Published
2015

39. Lactic acid is a sperm motility inactivation factor in the sperm storage tubules

Author

Matsuzaki Mei, Mizushima Shusei, Hiyama Gen, Hirohashi Noritaka, Shiba Kogiku, Inaba Kazuo, Suzuki Tomohiro, Dohra Hideo, Ohnishi Toshiyuki, Sato Yoshikatsu, Kohsaka Tetsuya, Ichikawa Yoshinobu, Atsumi Yusuke, Yoshimura Takashi, Sasanami Tomohiro, Matsuzaki Mei, Mizushima Shusei, Hiyama Gen, Hirohashi Noritaka, Shiba Kogiku, Inaba Kazuo, Suzuki Tomohiro, Dohra Hideo, Ohnishi Toshiyuki, Sato Yoshikatsu, Kohsaka Tetsuya, Ichikawa Yoshinobu, Atsumi Yusuke, Yoshimura Takashi, and Sasanami Tomohiro

Abstract

publisher

Published
2015

40. Lactic acid is a sperm motility inactivation factor in the sperm storage tubules

Author

Matsuzaki, Mei, primary, Mizushima, Shusei, additional, Hiyama, Gen, additional, Hirohashi, Noritaka, additional, Shiba, Kogiku, additional, Inaba, Kazuo, additional, Suzuki, Tomohiro, additional, Dohra, Hideo, additional, Ohnishi, Toshiyuki, additional, Sato, Yoshikatsu, additional, Kohsaka, Tetsuya, additional, Ichikawa, Yoshinobu, additional, Atsumi, Yusuke, additional, Yoshimura, Takashi, additional, and Sasanami, Tomohiro, additional

Published
2015
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41. A unique mechanism of successful fertilization in a domestic bird

Author

Sasanami, Tomohiro, primary, Izumi, Shunsuke, additional, Sakurai, Naoki, additional, Hirata, Toshifumi, additional, Mizushima, Shusei, additional, Matsuzaki, Mei, additional, Hiyama, Gen, additional, Yorinaga, Eriko, additional, Yoshimura, Takashi, additional, Ukena, Kazuyoshi, additional, and Tsutsui, Kazuyoshi, additional

Published
2015
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46. Wnt signaling blockade is essential for maintaining the pluripotency of chicken embryonic stem cells

Author

Kajihara, Ryota, Ezaki, Ryo, Ichikawa, Kennosuke, Watanabe, Tenkai, Terada, Takumi, Matsuzaki, Mei, and Horiuchi, Hiroyuki

Abstract

Activation of Wnt/β-catenin signaling supports the self-renewal of mouse embryonic stem cells. We aimed to understand the effects of Wnt signaling activation or inhibition on chicken embryonic stem cells (chESCs), as these effects are largely unknown. When the glycogen synthase kinase-3 β inhibitor CHIR99021—which activates Wnt signaling—was added to chESC cultures, the colony shape flattened, and the expression levels of pluripotency-related (NANOG, SOX2, SOX3, OCT4, LIN28A, DNMT3B, and PRDM14) and germ cell (CVHand DAZL) markers showed a decreasing trend, and the growth of chESCs was inhibited after approximately 7 d. By contrast, when the Wnt signaling inhibitor XAV939 was added to the culture, dense and compact multipotent colonies (morphologically similar to mouse embryonic stem cell colonies) showing stable expression of pluripotency-related and germline markers were formed. The addition of XAV939 stabilized the proliferation of chESCs in the early stages of culture and promoted their establishment. Furthermore, these chESCs formed chimeras. In conclusion, functional chESCs can be stably cultured using Wnt signaling inhibitors. These findings suggest the importance of Wnt/β-catenin signaling in avian stem cells, offering valuable insights for applied research using chESCs.

Published
2023
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47. Knock-in of the duck retinoic acid-inducible gene I (RIG-I) into the Mxgene in DF-1 cells enables both stable and immune response-dependent RIG-I expression

Author

Ichikawa, Kennosuke, Motoe, Yuzuha, Ezaki, Ryo, Matsuzaki, Mei, and Horiuchi, Hiroyuki

Abstract

Waterfowls, such as ducks, are natural hosts of avian influenza virus (AIV) and can genetically limit the pathogenicity. On the other hand, some AIV strains cause severe pathogenicity in chickens. It is suggested that differences in the pathogenicity of AIV infection between waterfowls and chickens are related to the expression of retinoic acid-inducible gene I (RIG-I), a pattern recognition receptor that chickens evolutionally lack. Here, we knocked-in the duck RIG-Ibearing the T2A peptide sequence at the 3′ region of the Mx, an interferon-stimulated gene (ISG), in chicken embryo fibroblast cells (DF-1) using the precise integration into target chromosome (PITCh) system to control the duck RIG-I expression in chickens. The expression patterns of the duck RIG-Iwere then analyzed using qPCR. The knocked-in DF-1 cells expressed RIG-I via the stimulation of IFN-β and poly(I:C) in a dose-dependent manner. Moreover, poly(I:C) stimulation in the knocked-in DF-1 cells upregulated RIG-I-like receptor (RLR) family signaling pathway-related genes IFN-β, OASL,and IRF7. The IFN-β-dependent expression of RIG-I and upregulation of IFN-βin the poly(I:C) stimulation demonstrated a positive-feedback loop via RIG-I, usually evident in ducks. Overall, this novel strategy established RIG-I-dependent immune response in chickens without overexpression of RIG-I and disruption of the host genes.

Published
2021
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48. Sperm activation by heat shock protein 70 supports the migration of sperm released from sperm storage tubules in Japanese quail (Coturnix japonica).

Author

Hiyama G, Matsuzaki M, Mizushima S, Dohra H, Ikegami K, Yoshimura T, Shiba K, Inaba K, and Sasanami T

Subjects
Animals, Antibodies administration & dosage, Female, Fertilization drug effects, Fertilization in Vitro drug effects, Gene Expression, HSP70 Heat-Shock Proteins genetics, HSP70 Heat-Shock Proteins immunology, Male, Oviducts chemistry, Oviposition, Ovulation, RNA, Messenger analysis, Sperm Motility drug effects, Spermatozoa chemistry, Uterus drug effects, Voltage-Dependent Anion Channel 2 physiology, Coturnix, HSP70 Heat-Shock Proteins pharmacology, Oviducts physiology, Sperm Transport physiology, Spermatozoa drug effects, Spermatozoa physiology
Abstract

Systems for maintaining the viability of ejaculated sperm in the female reproductive tract are widespread among vertebrates and invertebrates. In birds, this sperm storage function is performed by specialized simple tubular invaginations called sperm storage tubules (SSTs) in the uterovaginal junction (UVJ) of the oviduct. Although the incidence and physiological reasons for sperm storage in birds have been reported extensively, the mechanisms of sperm uptake by the SSTs, sperm maintenance within the SSTs, and control of sperm release from the SSTs are poorly understood. In this study, we demonstrated that the highly conserved heat shock protein 70 (HSP70) stimulates sperm motility in vitro and also that HSP70 expressed in the UVJ may facilitate the migration of sperm released from the SSTs. Quantitative RT-PCR analysis demonstrated that the expression of HSP70 mRNA in the UVJ increases before ovulation/oviposition. Gene-specific in situ hybridization and immunohistochemical analysis with a specific antibody to HSP70 demonstrated that HSP70 is localized in the surface epithelium of the UVJ. Furthermore, injection of anti-HSP70 antibody into the vagina significantly inhibited fertilization in vivo. In addition, we found that recombinant HSP70 activates flagellar movement in the sperm and that the binding of recombinant HSP70 to the sperm surface is mediated through an interaction with voltage-dependent anion channel protein 2 (VDAC2). Our results suggest that HSP70 binds to the sperm surface by interacting with VDAC2 and activating sperm motility. This binding appears to play an important role in sperm migration within the oviduct.

Published
2013
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