• Title/Summary/Keyword: Nucleus condensation

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Cytochemical Localization of Nuclear Actin of Sperm and Spermatids in Urechis unicinctus

  • Shin, Kil-Sang;Kim, Ho-Jin;Kwon, Hyuk-Jae;Kim, Wan-Jong
    • Animal cells and systems
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    • v.9 no.2
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    • pp.65-73
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    • 2005
  • In this study, we found that sperm ball of Urechis unicinctus consisted of a somatic cell and spermatogenic cells. After separation from the sperm ball, individual spermatid floated freely in the coelomic fluid and differentiated into a mature sperm. Because of many nuclear vacuoles, spermatid nucleus was observed to be heterogeneous. Later, the spermatid nucleus condensed into the homogeneous round nucleus of the mature sperm. Perinuclear microtubules could be seen but did not seem to be organized into manchette microtubules. To understand the nature of nuclear condensation during spermiogenesis, the sperm and spermatids (spermiogenic cells) were treated with FITC-phalloidin, or anti-actin-FITC, or labeled with antiactin immunogold particles (AAIP; 10 nm) followed by transmission electron microscopy or confocal laser scanning microscopy. The anti-actin-FITC and FITC-phalloidin reactions occurred distinctly in the nuclei of both spermiogenic cells. FITC-phalloidin reacted more intensely with acrosomes. The AAIP were incorporated mainly into nuclei of both cells sometimes showing local distribution in the nucleus. Nuclear vacuoles of spermatids disappeared progressively with condensation of the nucleus, as the number of incorporated $AAIP/{\mu}m^2$ increased. These results suggest that nuclear actin microfilaments might be closely related to nuclear condensation.

Effect of Electrical Preactivation of Recipient Cytoplasm on Nuclear Remodelling in Nuclear Transplant Rabbit Embryos (수핵란의 전 활성화가 토끼 핵이식 수정란의 핵 재구성에 미치는 효과)

  • 전병균;김윤연;정기화;곽대오;이효종;최상용;박충생
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.229-238
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    • 1997
  • Chromosome condensation and swelling of the donor nucleus have been known as the early morphological indicators of chromatin remodelling after injection of a foreign nucleus into an enucleated recipient cytoplasm. The effects of non-preactivation and electrical preactivation of recipient cytoplasm, prior to fusing a donor nucleus, on the profile of nuclear remodelling in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G1 phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgical procedure. The separated G1 phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation and the separated G1 phase blastomeres of 32-cell stage were injected. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The nuclei of nuclear transplant embryos fused into non-preactivated and/or preactivated recipient cytoplasm were stained by Hoechst 33342 at 0, 1.5, 2, 4, 6, 8, 10 hrs post-fusion and were observed under an fluorescence microscopy. Accurate measurements of nuclear diameter were revealed with an ocular micrometer at 200$\times$. Upon blastomere fusion into non-preactivated recipient cytoplasm, a prematurely chromosome condensation at 1.5 hrs post-fusion and nuclear swelling at 8 hrs post-fusion were occurred as 91.6% and 86.1%, respectively. But the nuclei of nuclear transplant embryos fused into preactivated recipient cytoplasm, as o, pp.sed to non-preactivated recipient cytoplasm, were not occurred chromosome condensation and extensive nuclear swelling. Nuclear diameter fused into non-preactivated and preactivated recipient cytoplasm at hrs post-fusion was 30.2$\pm$0.74 and 15.2$\pm$1.32${\mu}{\textrm}{m}$, respectively. These results indicated that onset of unclear condensation and swelling which was associated with oocytes activation were critical steps in the process of chromatin swelling. Futhermore, complete reprogramming seemed only possible after remodelling of the donor nucleus by chromosome condensation and nuclear swelling.

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CRM646-A, a Fungal Metabolite, Induces Nucleus Condensation by Increasing Ca2+ Levels in Rat 3Y1 Fibroblast Cells

  • Asami, Yukihiro;Kim, Sun-Ok;Jang, Jun-Pil;Ko, Sung-Kyun;Kim, Bo Yeon;Osada, Hiroyuki;Jang, Jae-Hyuk;Ahn, Jong Seog
    • Journal of Microbiology and Biotechnology
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    • v.30 no.1
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    • pp.31-37
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    • 2020
  • We previously identified a new heparinase inhibitor fungal metabolite, named CRM646-A, which showed inhibition of heparinase and telomerase activities in an in vitro enzyme assay and antimetastatic activity in a cell-based assay. In this study, we elucidated the mechanism by which CRM646-A rapidly induced nucleus condensation, plasma membrane disruption and morphological changes by increasing intracellular Ca2+ levels. Furthermore, PD98059, a mitogen-activated protein kinase (MEK) inhibitor, inhibited CRM646-A-induced nucleus condensation through ERK1/2 activation in rat 3Y1 fibroblast cells. We identified CRM646-A as a Ca2+ ionophore-like agent with a distinctly different chemical structure from that of previously reported Ca2+ ionophores. These results indicate that CRM646-A has the potential to be used as a new and effective antimetastatic drug.

Investigation of Particle Generation in a Hard Disk Drive During the Start/Stop Period (구동 및 정지시 하드 디스크 드라이브 내 입자 발생 특성에 관한 실험적 연구)

  • Park, Hee-Sung;Yoo, Yong-cheol;Bae, Gwi-Nam;Hwang, Jung-Ho
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.23 no.6
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    • pp.744-752
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    • 1999
  • Particle contamination in a hard disk drive has been recognized as an important issue because particles can significantly affect the reliability of the drive. In this study, characteristics of the particle generation in a hard disk drive during the start/stop period were investigated. The number of particles generated in the drive was measured at 5 locations by using condensation nucleus counters(CNC) with sampling probes. It was found that most of particles were generated whenever the drive started or stopped. The number of particles detected by the condensation nucleus counter depended on sampling parameters including sampling location, shape of sampling probe, and sampling velocity.

Sperm Injection into Maturing and Activated Porcine Oocytes

  • Kim, Bong-Ki;Lee, Yun-Jung;Cui, Xiang-Shun;Kim, Nam-Hyung
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.41-41
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    • 2001
  • Chromatin configuration and microtubule assembly were determined in porcine maturing and activated oocytes following intracytoplasmic sperm injection. Microtubule localization was confirmed using a mouse monoclonal antibody to $\alpha$-tubulin and detected using a fluorescent labeled goat anti-mouse secondary antibody. DNA was stained with propidium iodide. The image of microtubules and chromatin was captured using laser scanning confocal microscope. In germinal vesicle stage oocyte, sperm chromatin remained condensation and sperm derived microtubules were not observed at 8 to 12 h after sperm injection. At 24 h after injection, the sperm nucleus developed to the metaphase chromatin along the metaphase structure of female nucleus. In some metaphase I stage oocytes, sperm chromatin decondensed at 8 h to 12 h after injection, sperm aster was seen soon after sperm injection. At 24 h after sperm injection into metaphase I stage oocyte, male chromatin developed to the metaphase chromatin while female chromatin extruded first polar body and formed the metaphase chromatin. At 12 to 15 h after sperm injection into preactivated oocytes, condensed sperm nucleus was located in close proximity of female pronucleus. However, the condensed nucleus did not fuse with female pronucleus. In preactivated ocytes, injected sperm remained condensation, a few sperm organized small microtubular aster. Instead, maternal derived microtubules were organized near the female chromatin, which seem to move condensed male chromatin near to the female pronucleus. These results suggest that sperm nuclear decondensing activity and nucleation activity of centrosome during fertilization are cell cycle dependent. In absence of male functional centrosome, female origin centrosome takes over the role of microtubule nucleation for nuclear movement.

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Observational Evidence of Giant Cloud Condensation Nucleus Effects on the Precipitation Sensitivity in Marine Stratocumulus Clouds

  • Jung, Eunsil
    • Journal of the Korean earth science society
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    • v.43 no.4
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    • pp.498-510
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    • 2022
  • Cloud-aerosol interactions are one of the paramount but least understood forcing factors in climate systems. Generally, an increase in the concentration of aerosols increases the concentration of cloud droplet numbers, implying that clouds tend to persist for longer than usual, suppressing precipitation in the warm boundary layer. The cloud lifetime effect has been the center of discussion in the scientific community, partly because of the lack of cloud life cycle observations and partly because of cloud problems. In this study, the precipitation susceptibility (So) matrix was employed to estimate the aerosols' effect on precipitation, while the non-aerosol effect is minimized. The So was calculated for the typical coupled, well-mixed maritime stratocumulus decks and giant cloud condensation nucleus (GCCN) seeded clouds. The GCCN-artificially introduced to the marine stratocumulus cloud decks-is shown to initiate precipitation and reduces So to approximately zero, demonstrating the cloud lifetime hypothesis. The results suggest that the response of precipitation to changes in GCCN must be considered for accurate prediction of aerosol-cloud-precipitation interaction by model studies

Spermiogenesis in the Korean Greater Horseshoe Bat, Rhinolophus ferrumequinum korai (한국산 관박쥐 (Rhinolophus ferrumequinum korai)에 있어서의 정자변태)

  • Lee, Jung-Hun;Choi, Byung-Jin;Son, Sung-Won
    • Applied Microscopy
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    • v.22 no.2
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    • pp.97-117
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    • 1992
  • In order to study process of spermiogenesis of the Korean greater horseshoe bat, Rhinolophus ferrumequinum korai, the cycle of seminiferous epithelium was examined by the light and electron microscope and the following results were obtained based on the epithelial cell differentiation. 1. Spermiogenesis occurred from early July to mid-Octber, and spermatogenic activity was vigorous from mid-August to late September. Spermatocytes including spermatogonia were found to be degenerated in only July. It is deduced that the degeneration serves as the mechanism to regulate effective use of energy to prepare for mating and hibernating periods, and regulation of breeding cycle. 2. Spermiogenesis of the Korean greater horseshoe bat was divided according to differentiation of the cell structure, into Golgi, cap, acrosome, maturation and spermiation phases; Golgi, cap and spermiation phases were further divided into two steps of early and late phase respectively, and acrosome phase into three steps of early, mid and late phases, and maturation phase has only one step. Hence, the spermiogenesis consists of ten phases. The first research was done in this article on the changes of chromatin with nucleus, the time of appearance and disappearance of chromatin granules, in case of Korean greater horseshoe bat (Rhinolophus ferrumequinum korai). Chromatin granule began to be condensed in late Golgi and the condensation proceeded to form an irregular mass of a electron-dense chromatin in a form of circular cylinder in the center of nucleus at the phase of maturation. Finally, the chromatin condensation proceeded and perfect nucleus of sperm with homogeneous density was formed when the sperm was separated from Sertoli cell. Therefore, appearance and disappearance of chromatin granules occurred in the period of time between late Golgi and maturation phase, The tail of sperm began to develop in early cap phase, Numerous lipid droplets were obseved in the cytoplasm of spermatids during the maturation phase, which seemed to be used as energy source necessary to make mature sperm during spermiogenesis.

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Spermatogenesis of Siamese Fighting Fish, Betta splendens, Osphronemidae, Teleostei

  • Lim, Sung Ha;Koh, Yeong Kyeong;Chang, Byung Soo;Kim, Dong Heui
    • Applied Microscopy
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    • v.44 no.1
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    • pp.1-7
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    • 2014
  • The spermatogenesis of Siamese fighting fish, Betta splendens, belongs to Osphronemidae was investigated by light and electron microscopic observations. In primary spermatocyte stage, the nucleus was comparatively large ellipsoidal, and mitochondria showed a marked development in cytoplasm. In secondary spermatocyte stage, the germ cells were smaller than that of primary spermatocytes. The nucleus was a spherical shape and intercellular space was formed between germ cells. In spermatid stage, the early spermatids were not much different from a secondary spermatocyte. But, the chromatin condensation was occurred from the outside to the inside. The nucleus was more condensed. Intracellular space was larger than early spermatid. The mitochondria were rearranged in a middle piece, and occupied about half of the head part in early sperm. In sperm stage, the head of mature sperm was a spherical shape and had no acrosome. The flagellum was showed the typical 9+2 array of microtubules. Also, the tail of sperm had no lateral fins and outer coarse fibers. These ultrastructural characteristics can be used in classification of species.

Apoptic and antimetastatic effects of ursolic acid isolated from Oldenlandia diffusae Herba (백화사설초(白花蛇舌草)로부터 분리(分離)한 ursolic acid의 자연살해효과(自然殺害效果)와 항전이작용(抗轉移作用))

  • Kim, Sung-Hoon
    • Journal of Haehwa Medicine
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    • v.5 no.2
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    • pp.523-533
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    • 1997
  • Ursolic acid(UA) was isolated from Oldenlandiae diffusae Herba, one of the commonly used medicinal herbs for the treatment of cancer. IC50 of UA against cancer cell lines as SNU-1, B16-Fo. SK-OV3, HCT15, XF498, SK-MEL and A549 was $6{\mu}g/ml$, 4$4.4{\mu}g/ml$, $4.5{\mu}g/ml$, $4.6{\mu}g/ml$ and $4.2{\mu}g/ml$ respectively suggesting cytotoxicity against cancer cells. DNA fragmentation was expressed from the concnetration of $5.5{\mu}g/ml$ of UA by agarose electrphoresis. In the observation of morphological changes by phase contrast microscope, SEM and TEM, cell injury and condensation of cytoplasm from nucleus began 4 hr after UA treatment. fragmentaion of nucleus and injury of cell membrane was shown 24 hr after UA treatmeilt with SNU-1 cells. Aurin tricarboxic acid as endonuclease inhibitor. and nicotinamide as poly(ADP-ribose) polymerase inhibitor protected over 50% of cytotoxicity of UA against SNU-1 was at the concentrations of $3{\mu}M$ and $300{\mu}M$ respectively suggesting UA acts on nucleus. These results suggest that UA had antimetastatic effect and induced apoptosis.

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