• 제목/요약/키워드: fresh semen

검색결과 94건 처리시간 0.021초

Characterization of Mouse Interferon-Induced Transmembrane Protein-1 Expression in Mouse Testis

  • Lee, Ran;Park, Hyun Jung;Lee, Won Young;Kim, Ji Hyuk;Kim, In Chul;Kim, Dong Woon;Lee, Sung Dae;Jung, Hyun Jung;Kim, Jong Moon;Yoon, Hyung Moon;Kwon, Hyuk Jung;Song, Hyuk
    • Reproductive and Developmental Biology
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    • 제36권3호
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    • pp.225-230
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    • 2012
  • Interferon induced transmembrane protein-1 (Ifitm-1) has been reported to have an important role in primordial germ cell formation, and it has expressed in female reproductive organ. In the present study, Ifitm-1 gene expression was identified in testes and all part of epididymis using western immunoblot and immunohistochemistry. Interestingly, Ifitm-1 expression was observed on the head of spermatozoa. To investigate the role of Ifitm-1 gene expression in behavior of spermatozoa after acrosome reaction, fresh sperm was incubated with calcium ionophore to induce acrosome reaction, whereas the expression of Ifitm-1 was not altered after the acrosome reaction. Then to identify the effect of Ifitm-1 in sperm motility and other seminal parameters, different concentration of Ifitm-1 antibody was incubated with spermatozoa, and seminal parameters were assessed using computer-assisted semen analysis (CASA). Interestingly, motility, progressive, and VAP were increased in the sperm with Ifitm-1 antibody treated compared to rabbit serum, however other parameters such as straightness were not changed. In order to identify the functional significance of Ifitm-1 in fertilization, capacitated spermatozoa were pre-incubated with anti-Ifitm-1 antibody and subsequently examined the ability to adhere to mouse oocytes. However, any defection or alteration in sperm-egg fusion was not found, Ifitm-1 antibody treated or non-treated spermatozoa showed a normal penetration. Although the precise role of Ifitm-1 in sperm motility and following fertilization need to be elucidated, this study suggests that the activation of Ifitm-1 on the sperm may enhance the motility of spermatozoa in mice.

Acrosome staining with Coomassie brilliant blue G or R on the horse spermatozoa (Coomassie brilliant blue G 및 R을 이용한 말 정액 첨체 염색 기법)

  • Kim, Sung Woo;Shin, Sang Min;Yu, Yeonhee;Lee, Jae-Yeong;Kim, Chan-Lan;Ko, Yeoung-Gyu
    • Journal of the Korea Academia-Industrial cooperation Society
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    • 제21권9호
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    • pp.57-63
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    • 2020
  • To develop simple acrosome staining of horse spermatozoa, this study tested the binding properties of Coomassie brilliant blue G or R on the sperm smears after 3.7% paraformaldehyde (PF) or 35% methanol (MT) fixation. After being fixed with PF and stained with 0.05, 0.1, or 0.2 % of CBB G or R for 2 min, horse spermatozoa were examined for their intact acrosome status. The intact acrosome of fresh horse spermatozoa were 62.6% and 61.5% with 0.05% of the G and R CBB solution, but 80.2 and 79.7% with G type and 78.1 and 76.0% with R type. On the other hand, when MT was used for fixation, the acrosome reacting sperm ratio was 3.5%, but was 9.0% in the case of PF. These results show that the intact acrosome of horse sperm could be judged using a 0.1~0.2% CBB G or R staining technique. PF would be an essential fixative for examining acrosome reacting horse spermatozoa. This method could be used to identify sperm with a damaged acrosome during low-temperature storage or cryopreservation for artificial insemination of horses.

Influence of the Substrate and Inhibitors Related to Phosphatidylinositol Metabolism in the Maturation Processes of Porcine Oocytes (돼지 난모세포의 성숙과정에서 Phosphatidylinositol 대사의 기질 및 억제인자의 영향)

  • 강승률;양보석;조인철;이성수;정진관
    • Journal of Embryo Transfer
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    • 제16권2호
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    • pp.91-98
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    • 2001
  • We evaluated the effects of the substrate and inhibitors related to phosphatidylinositol metabolism on in vitro maturation and fertilization of porcine oocytes. Cumulus-oocyte complexes were cultured in mTLP-PVA medium supplemented with or without inositol (250 mM) fur 46h. Subsequently, these oocytes were inseminated with fresh boar semen in mTALP-PVA medium for 6h. At 6h after insemination, oocytes were cultured for further 12 h in TCM-199 supplemented with 10% FBS (fetal bovine serum). The higher percentage of oocytes in inositol-supplemented medium reached metaphase of the second meiotic division compared to those in control (81.4% vs. 67.3%; P<0.()5). following 18 h of insemination, more number of male pronuclei were formed in the oocytes matured in inositol-supplemented medium than in those of control experiment (42.0% vs. 27.3%; P<0.05). When oocytes were cultured in medium with 10mM LiCl (chloride lithium) or 0.5mM dbcAMP (dibutyryl cyclic adenosine monophosphate) to determine the role of inositol on the maturation of oocytes, these two drugs inhibited the meiotic division of oocytes (P<0.05). However, addition of inositol to the culture medium did overcome the inhibitory effect of these drugs on the oocyte maturation. DbcAMP and verapamil supplemented synergistically arrested the meiotic division of oocytes. Addition of verapamil did not inhibit germinal vesicle breakdown, but it severly inhibited the second meiotic division of oocytes. These results suggest that inositol exert its improving effects on maturation, by activating the PI (phosphatidylinositol) cycle and causing beneficial changes in both cytoplasm and membrane of oocytes.

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TLR-1, TLR-2, and TLR-6 MYD88-dependent signaling pathway: A potential factor in the interaction of high-DNA fragmentation human sperm with fallopian tube epithelial cells

  • Zahra Zandieh;Azam Govahi;Azin Aghamajidi;Ehsan Raoufi;Fatemehsadat Amjadi;Samaneh Aghajanpour;Masoomeh Golestan;Reza Aflatoonian
    • Clinical and Experimental Reproductive Medicine
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    • 제50권1호
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    • pp.44-52
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    • 2023
  • Objective: The DNA integrity of spermatozoa that attach to fallopian tube (FT) cells is higher than spermatozoa that do not attach. FT epithelial cells can distinguish normal and abnormal sperm chromatin. This study investigated the effects of sperm with a high-DNA fragmentation index (DFI) from men with unexplained repeated implantation failure (RIF) on the Toll-like receptor (TLR) signaling pathway in human FT cells in vitro. Methods: Ten men with a RIF history and high-DFI and 10 healthy donors with low-DFI comprised the high-DFI (>30%) and control (<30%) groups, respectively. After fresh semen preparation, sperm were co-cultured with a human FT epithelial cell line (OE-E6/E7) for 24 hours. RNA was extracted from the cell line and the human innate and adaptive immune responses were tested using an RT2 profiler polymerase chain reaction (PCR) array. Results: The PCR array data showed significantly higher TLR-1, TLR-2, TLR-3, TLR-6, interleukin 1α (IL-1α), IL-1β, IL-6, IL-12, interferon α (IFN-α), IFN-β, tumor necrosis factor α (TNF-α), CXCL8, GM-CSF, G-CSF, CD14, ELK1, IRAK1, IRAK2, IRAK4, IRF1, IRF3, LY96, MAP2K3, MAP2K4, MAP3K7, MAP4K4, MAPK8, MAPK8IP3, MYD88, NFKB1, NFKB2, REL, TIRAP, and TRAF6 expression in the high-DFI group than in the control group. These factors are all involved in the TLR-MyD88 signaling pathway. Conclusion: The MyD88-dependent pathway through TLR-1, TLR-2, and TLR-6 activation may be one of the main inflammatory pathways activated by high-DFI sperm from men with RIF. Following activation of this pathway, epithelial cells produce inflammatory cytokines, resulting in neutrophil infiltration, activation, phagocytosis, neutrophil extracellular trap formation, and apoptosis.