• Clinical and Experimental Reproductive Medicine
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• v.28 no.1
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• pp.1-12
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• 2001

Objective: The present study was done to clarify the effects of nicotine and nicotine tartrate on the mouse oocyte maturation in vitro. Methods: GV (germinal vesicle) oocytes were isolated from Graafian follicle of ovaries with sharp needles under a stereomicroscope from female mouse of ICR strain (4 weeks old). Collected oocytes were cultured for 17 hours at $37^{\circ}C$, 5% $CO_2$ in air and 100% humidified condition in incubator. New MHBS was the basic medium used in which nicotine, nicotine tartrate, and mecamylamine (antagonist of nicotinic acetylcholine receptor) were added depending on the experimental group. GV oocytes were cultured in one of these media. Results: Nicotine ($300{\mu}M{\sim}5mM$) had no effects on GVBD (germinal vesicle breakdown) compared to the control, but increasing concentration of nicotine led to an decrease in the first polar body formation. However, nicotine ($10{\sim}500{\mu}M$) induced GVBD in a dose-dependent manner of GV oocytes in a medium containing dbcAMP. Nicotine tartrate ($50{\mu}M{\sim}5mM$) had no effects on GVBD compared to the control but, increasing concentration of nicotine tartrate led to an decrease in the first polar body formation. Mecamylamine $10{\mu}M$ added to the medium containing nicotine ($300{\mu}M{\sim}5mM$) showed higher percentage of the first polar body formation compared to the nicotine ($300{\mu}M{\sim}5mM$) treatment group. Mecamylamine $10{\mu}M$ added to the medium containing nicotine tartrate ($50{\mu}M{\sim}5mM$) showed higher percentage of the first polar body formation compared to the nicotine tartrate ($50{\mu}M{\sim}5mM$) treatment group. Conclusion: The present study suggest that nicotine and nicotine tartrate have the harmful effects on the meiotic maturation of the mouse oocytes in vitro. However, mecamylamine block harmful effects of nicotine and nictine tartrate.

• Development and Reproduction
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• v.18 no.3
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• pp.173-178
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• 2014

In order to monitor the developmental features of embryos, larvae, and juveniles of Oryzias latipes (Temminck and Schlegel), Oryzias latipes was caught in river of Shinduck-dong, Yeosu-si, Jeollanam-do, on May 2011, and experiments were carried out in Ichthyology laboratory at Chonnam National University. The blastodisc step was the first level for natural spawning. The optic vesicle, Kupffer's vesicle, myotome began to appear 75 hours 57 minutes later. After blastodisc development, the pectoral fins were made at 143 hours 37 minutes and the tail was separated started at the same time. Hatching was observed at 167 hours 27 minutes after blastodisc. The total length of the hatched larvae was 4.95~5.10 mm (mean, 5.01 mm), the mouth and anus were opened. Larvae used yolk completely after 3 days after hatching. The total length larvae was 5.45~5.56 mm (mean, 5.52 mm) after 8 days after hatching, and appeared the stems for tail. The stems pectoral, anal fin were showed after 14 days and the stems dorsal, ventral fin were appeared after 19 days. For 35 days after hatching, the total length of larvae 13.95~15.30 mm (mean, 14.64 mm), and at this time, fins and body were transferred like the adult Oryzias latipes.

• Applied Chemistry for Engineering
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• v.28 no.6
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• pp.679-684
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• 2017

Hydrated liquid crystalline vesicles incorporating a edge activator, which confers flexibility to the vesicle membranes, were prepared and niacinamide was encapsulated in them. The formation of liquid crystalline phases and their thermal phase transitions were investigated by polarized optical microscopy and differential scanning calorimetry (DSC), respectively. Droplet sizes of the vesicles were reduced to several tens of nanometers by incorporating edge activators, such as sodium deoxycholate, lysolecithin, or polysorbate 80. The amount of niacinamide permeated into a pig skin increased greatly using the hydrated liquid crystalline vesicles compared to the case where niacinamide was applied in an aqueous solution state. The vesicles incorporating 10% sodium deoxycholate increased the amount of niacinamide permeated nearly four times. These results suggest that edge activators are effective in improving the skin permeability of vesicles.

• Applied Microscopy
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• v.25 no.3
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• pp.63-74
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• 1995

The development of pale chub oocyte from the immature oogonium to mature oocyte was investigated by light and electron microscope. The cytoplasm of pale chub oogonia was acidic and many vesicles were located at inner side of nuclear membrane. In primary oocytes, yolk vesicles were distributed in cytoplasm. Also, fibrous materials and protuberances were distributed on the surface of zona radiata. The nucleus of secondary oocyte was enlarged and yolk vesicles in cytoplasm migrated to zona radiata. In early egg, yolk mass are formed and yolk vesicles were located at inner side of zona radiata. Three-layered zona radiata was about $3{\mu}m$ in thickness. The three layers were an outer fibrous material layer, a middle nurse cell layer in which microvilli of early egg cytoplasm contact with processes of nurse cells, and an inner layer with high electron density. In mature egg, euchromatin and a germinal vesicle were developed, mitochondria, free ribosomes, and yolk mass were distributed in cytoplasm. But, yolk vesicles were disappeared. Specially, zona radiata of matured eggs were better thin than the one of immature eggs In conclusion, it is summerized that the oogenesis of pale chub were the increase of cell size, the formation and accumulation of yolk, the decrease in nucleat electron density, changes of zona radiata, and the development of microvilli.

• Applied Microscopy
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• v.34 no.3
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• pp.159-170
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• 2004

Spermiogenesis in the Korean squirrel, Tamias sibiricus, was investigated by transmission electron microscopy. Spermiogenesis was divided into Golgi, cap, acrosome, maturation and spermiation phases based on the characteristics of acrosomal changes and nuclear shape. Beside, the Golgi, cap and acrosomal steps were subdivided into three phases of early, middle and late phase respectively, the maturation step was divided into two phases of early and late phase, and spermiation step has only one phase. Thus, the spermiognesis of T. sibiricus was divided into a total of twelve phases. In Golgi phase (steps 1-3), a well developed Golgi complex was located close to the vesicles, the acrosomal vesicle fixd to a recess of nuclear membrane at step 3. During cap phase (steps 4-6), the acrosomal vesicle spred over the nuclear surface to cover a third of the nucleus, and the acrosomal granule was not yet flattened. At acrosomal phase (steps 7-9), the nucleus and acrosome were elongated but nucleoplasm was not condensed. During maturation phase (steps 10-11), the nucleoplasm was more condensed, and the mitochondria completely arranged the center of axoneme. The spatulate-sperm head was completely formed at spermiation phase (step 12).

• Applied Microscopy
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• v.34 no.4
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• pp.223-230
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• 2004

To identify the structural basis of mutations that affect synaptic transmission we have begun quantitative ultrastructural descriptions of synapses in cultured Drosophila embryonic neurons. In wild-type cultures, synapses are distinguished by the parallel arrangement of a thickened pre- and post synaptic membrane separated by a synaptic cleft. The presynaptic active zones and postsynaptic densities are defined by electron dense material close to the membrane. Presynaptic regions are also characterized by the presence of one or more electron dense regions, presynaptic densities, around which a variable number of small, clear core synaptic vesicles (mean $35.1{\pm}1.44$ nm in diameter) are clustered. Subsets of these vesicles are in direct contact with either the presynaptic density or the membrane and are considered morphologically docked. A small number of larger, dense core vesicles are also observed in most presynaptic profiles.

• Parasites, Hosts and Diseases
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• v.35 no.2
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• pp.95-104
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• 1997

The localization of worm antigen of Neodiplostomum seoulere was examined by immuno-electronmicroscopic observation. Not only the immunized serum of mice with crude worm extract of N. seoulene but also serum of infected mouse were reacted to the worm section. Using immunized serum as primary antibody. the gold particles were deposited on the rough endoplasmic reticulum of the cell of tribocytic organ, spermatozoa in the seminal vesicle, microvilli of the caecum and vitelline follicle. Using infected serum, gold particles were deposited only on the vitelline follicle prominently. This finding suggested that the tribocytic organ, seminal vesicle, caeca and vitelline follicles may play a role of antigen to immunized serum with crude worm extract of N. seoulense, whereas the vitelline follicle, to the infected serum.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.10
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• pp.5063-5067
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• 2012

New molecular markers of cancer had emerged with novel applications in cancer prevention and therapeutics, including for breast cancer of unknown causes, which has a high impact on the health of women worldwide. The purpose of this research was to detemine protein and mRNA expression of synaptic vesicle 2 (SV2) isoforms A, B and C in breast cancer cell lines. Cultured cell lines MDA-MB-231, SKBR3, T47D were lysed and their protein and mRNA expression analyzed by real-time PCR and western blot technique, respectively. SV2A, B proteins were identified in non-tumor (MCF-10A) and tumor cell lines (MDA-MB-231 and T47D) while SV2C only was found in the T47D cell line. Furthermore, the genomic expression was consistent with protein expression for a such cell line, but in MDA-MB-231 there was no SV2B genomic expression, and the SV2C mRNA and protein were not found in the non tumoral cell line. These findings suggest a possible cellular transdifferentiation to neural character in breast cancer, of possible relevance to cancer development, and point to possible use of SV2 as molecular marker and a vehicle for cancer treatment with botulinum toxin.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.1
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• pp.41-47
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• 2017

Perhaps the most common type of reproductive dysfunction in captive fish is failure of females to undergo final oocyte maturation and thus to ovulate and spawn. The success of aquaculture could therefore be improved by developing techniques to enhance natural spawning, artificial maturation, and/or to induce ovulation in farmed fish. This study aimed to investigate the effects of prostaglandin $E_2$ ($PGE_2$) and prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$) on in vitro oocyte maturation (germinal vesicle breakdown, GVBD) and ovulation in the marine fish Chasmichthys dolichognathus. Post-vitellogenic follicles (0.80-0.94 mm diameter oocytes) were incubated with $PGE_2$ or $PGF_{2{\alpha}}$ at concentrations of 5, 50, or 500 ng/mL for 24 hours. A significant increase in GVBD was seen in 0.84 mm and 0.94 mm oocytes incubated with 50 ng/mL $PGE_2$ compared with the control. There was no significant increase in GVBD in any of the other experimental conditions (5 or 500 ng/mL $PGE_2$ or 5, 50, or 500 ng/mL $PGF_{2{\alpha}}$). Neither of the prostaglandins induced ovulation at the concentrations tested.These results suggest that GVBD was induced by incubation with 50 ng/mL $PGE_2$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.37 no.3
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• pp.192-196
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• 2004

The in vitro effect of bisphenol A (BPA) on ovarian steroidogenesis of the longchin goby (Chasmichthys dolichognathus) was investigated. Oocytes taken during the maturing phase (vitellogenic, fully vitellogenic or germinal vesicle breakdown stage) were incubated with BPA (100 ng/mL) in the presence of exogenous precursor $^{3}H-17\alpha\;hydroxyprogesterone\;(^{3}H-17\alphaOHP).$ Steroids were extracted from the media and the isolated oocytes, and the extracts were separated and identified by thin layer chromatography and gas chromatography-mass spectrometry. The identities of the major metabolites were progestogens $[17{\alpha}-hydroxy,20{\alpha}-dihydroprogesterone\;(17{\alpha}20{\alpha}OHP)\;and\;17{\alpha}-hydrxy,20{\beta}-dihydroprogesterone\;(17{\alpha}20{\beta}OHP),$ androgens [androstenedione (A4) and testosterone (T)] and estrogens [estrone $(E_1)\;and\;estradiol-17{\beta}(E_2)].$ BPA treatment inhibited production of estrogens in all the maturing phases and progestogens in the germinal vesicle migrating stage. Percentage yield of estrogens was decreased with increased yield of androgens. In conclusion, BPA had an inhibitory effect on the conversion of $^3H-17\alphaOHP$ to estrogens and progestogens. These results demonstrate that BPA can act either estrogenic or anti-estrogenic effects.