• Applied Microscopy
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• v.25 no.1
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• pp.1-14
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• 1995

The purpose of this study was to investigate the main sensory trigeminal nucleus in the aging rat brain by means of electron microscope. Male Sprague-Dawley rats, two (control group) and thirty six (aging group) months of age, were used. These animals were sacrificed by perfusion fixation with 2.5% glutaraldehyde-2.0% paraformaldehyde (0.1M phosphate buffer, pH 7.4) under sodium pentobarbital. The objective area was punched out with a sharp-edged metal cylinder of 0.8 mm in diameter. These blocks of tissue were then washed in 0.1M phosphate buffer, postfixed in 2% osmium tetroxide, dehydrated in a graded series of ethyl alcohol, and embedded in Epon 812. Thin sections were cut with Super Nova ultramicrotome, pick up on grids and double stained with lead citrate and uranyl acetate, and observed in JEOL 100B electron microscope. The results were as follows: 1. In the control group, the neuronal cell body of the main sensory trigeminal nucleus was filled with nucleus, Golgi complex, Nissl substance, mitochondria, microfilaments and microtubules. However, few Nissl substances are seen in neuronal cell body. Axoaxonic synapse, axodendritic synapse, axosomatic synapse, axospinous synapse, myelinated and unmyelinated nerve fibers were well organized around cell bodies. Neurons with abnormal changes were not seen. 2. In the aging group, the neuronal cell body of the main sensory trigeminal nucleus contained large number of lipofuscin granules, dense body and swollen mitochondria. Terminal boutons contained glycogen, crystal-like vesicle and membranous indicating first signs of degeneration. The dendrites were found to be in synaptic contact with altered axon terminals. Frequently axons filled with dark axoplasn and splitted myelin sheath were noticed.

• Development and Reproduction
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• v.16 no.1
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• pp.59-67
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• 2012

Previously, we found that $Zap70$ (Zeta-chain-associated protein kinase) expressed in the mouse oocytes and played significant role in completion of meiosis specifically at MI-MII (metaphase I-II) transition. Microinjection of $Zap70$ dsRNA into the cytoplasm of germinal vesicle oocyte resulted in MI arrest, and exhibited abnormalities in their spindles and chromosome configurations. The purpose of this study was to determine the mechanisms of action of $Zap70$ in oocyte maturation by evaluating downstream signal networking after $Zap70$ RNAi (RNA interference). The probe hybridization and data analysis were used by Affymetrix Gene Chip Mouse Genome 430 2.0 array and GenPlex 3.0 (ISTECH, Korea) software, respectively. Total 1,152 genes were up (n=366) and down (n=786) regulated after $Zap70$ RNAi. Among those genes changed, we confirmed the expressional changes of the genes involved in the regulation of actin cytoskeleton and MAPK (mitogen-activated protein kinase) signaling pathway, since the phenotypes of $Zap70$ RNAi in oocytes were found in the changes in the chromosome separation and spindle structures. We confirmed the changes in gene expression in the actin skeletal system as well as in the MAPK signaling pathway, and concluded that these changes are main cause of the aberrant chromosome arrangement and abnormal spindles after $Zap70$ RNAi.

• Applied Microscopy
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• v.28 no.1
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• pp.73-82
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• 1998

To elucidate how microfilaments and vesicles participate in bile formation, the pericanalicular cytoplasms were observed in the liver of rats treated with taurocholic acid by deep etching with rapid freezing, and copmpared them with the findings on convensional thin sections. The microfilaments were identified around the bile canaliculi in the forms of core filaments of microvilli, filaments of pericanalicular web running in parallel to the border of bile canaliculi, and filaments on the junctional complex. In taurocholic acid-treated rats, microfilaments could be visualized around the bile canaliculi and along their borders. The microfilaments appeared to be installed to link to both the canalicular membrane and vesicles. Such specialized microfilaments are considered to participate in the translocation of vesicles in the pericanalicular cytoplasm. From the evidence, it is assumed that the microfilament induces the vesicles to transport and fuse to bile canalicull into which bile acids is secreted by exocytosis.

• Journal of the Korean Society of Fisheries and Ocean Technology
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• v.43 no.4
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• pp.320-328
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• 2007

Maturation and spawning of the Pacific cod, Gadus macrocephalus was investigated based on the samples captured in East Sea of Korea from April 2006 to July 2007. Gonadosomatic index began to increase in November, and reached maximum between December and January. After spawning it began to decrease from March. Reproductive season was estimated to November-February, with peak in January. Fecundity was proportional to the size of the female, with the clutch size varying from 753,985 eggs in the smallest female(TL＝58.6cm) to 9,311,520 eggs in the largest(TL＝101.0cm). Size at 50% sexual maturity(TL50%) determined from mature females was 56.3cm of TL. Annual reproductive cycles of this species could be divided into six successive stages; immature stage(March-September), nucleolus stage(September-October), yolk vesicle stage(October-November), vitellogenic stage (November-December), ripe stage and spent stage(December-February).

• Journal of Embryo Transfer
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• v.27 no.4
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• pp.265-270
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• 2012

In canine, oocytes are ovulated at the GV (germinal vesicle) stage and they have to fulfill maturation phase before reaching metaphase II stage. The efficiency of in vitro maturation is still very low. Therefore, the aim of this study was to investigate the effect of in vitro maturation on nuclear changes of immature canine oocytes recovered from different reproductive stages ovaries and different culture conditions. The oocytes were cultured in TCM-199 with supplement at 5% $CO_2$ and $38.5^{\circ}C$ for 72 h. The nuclear maturation of canine oocytes was evaluated with Hoechst 33342 stain under fluorescence microscope (Fig. 1). The results of this study detected differences in in vitro maturation rate between oocytes recovered from follicle status and non-follicle status ovaries. However, these differences were not significant as indicated in Table 1 and Fig. 2. In regard to the effect of culture condition with supplements, we did not found significant differences compared with control group (Table 2, Table 3). One of the reasons for this data could be the conditions that ovaries were exposed during slaughtering process or the long distant transportation of the ovaries. Although these data have not shown clearly significant differences results compared with control, furthermore the different reproductive status ovaries was beneficial for maturation of oocytes in vitro and can be a basic part of knowledge to improve in vitro maturation of canine oocytes.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.5
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• pp.513-519
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• 1999

Direct exposure of renal tubular brush-border membranes (BBM) to free cadmium (Cd) causes a reduction in phosphate (Pi) transport capacity. Biochemical mechanism of this reduction was investigated in the present study. Renal proximal tubular brush-border membrane vesicles (BBMV) were isolated from rabbit kidney outer cortex by Mg precipitation method. Vesicles were exposed to $50{\sim}200\;{\mu}M\;CdCl_2$ for 30 min, then the phosphate transporter activity was determined. The range of Cd concentration employed in this study was comparable to that of the unbound Cd documented in renal cortical tissues of Cd-exposed animals at the time of onset of renal dysfunction. The rate of sodium-dependent phosphate transport $(Na^+-Pi\;cotransport)$ by BBMV was determined by $^{32}P-Iabeled$ inorganic phosphate uptake, and the number of $Na^+-Pi$ cotransporters in the BBM was assessed by Pi-protectable $^{14}C-labeled$ phosphonoformic acid $([^{14}C]PFA)$ binding. The exposure of BBMV to Cd decreased the $Na^+-Pi$ cotransport activity in proportion to the Cd concentration in the preincubation medium, but it showed no apparent effect on the Pi-protectable PFA binding. These results indicate that an interaction of renal BBM with free Cd induces a reduction in $Na^+-Pi$ cotransport activity without altering the carrier density in the membrane. This, in turn, suggest that the suppression of phosphate transport capacity $(V_{max})$ observed in Cd-treated renal BBM is due to a reduction in $Na^+-Pi$ translocation by existing carriers, possibly by Cd-induced fall in membrane fluidity.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.3
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• pp.344-353
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• 2018

Objective: cAMP and mature promoting factor (MPF) play critical roles during the maturation of mammalian oocytes. The aim of this study was to produce the offspring from denuded oocytes (DOs) in mice by regulating cAMP and MPF. Methods: In this study, we used DOs at the germinal vesicle (GV) stage in mice and regulated levels of cAMP and MPF in DOs by adding Forskolin and PD166285 during in vitro maturation without follicle stimulating hormone and luteinizing hormone, respectively. Results: Combined use of $50{\mu}M$ Forskolin for 3 h and $2.5{\mu}M$ PD166285 for additional 21 h enhanced the developmental competence of DOs, maturation rate of DOs was $76.71%{\pm}4.11%$, blastocyst rate was $18.33%{\pm}4.44%$ after parthenogenetic activation (PA). The DOs could successfully be fertilized with sperm in vitro, cleavage rate was $17.02%{\pm}5.82%$ and blastocyst rate was $5.65%{\pm}3.10%$. Besides, 2-cell in vitro fertilization embryos from DOs produced 4 normal live offspring (4/34). Conclusion: The results confirmed that the combination of Forskolin and PD166285 can induce DOs to complete meiosis process and produce normal offspring.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.357-366
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• 1996

The present experiments aimed to investigate the metabolism of calcium during oocyte maturation in rat. The concentration of free calcium and calmodulin in oocytes was measured respectively by using of fluo-3/AM and FITC with microscope fluorescence spectrometer. The ultrastructural localization of calcium precipitates in oocytes was observed with the transmission electron microscope. Cumulus-free immature oocytes(GV-oocyte) were cultured in vitro through 15 hours. The free calcium concentration in GV oocyte was $55.9{\pm}3.5nM$. In calcium-containing medium, the free calcium concentration was increased in germinal vesicle breakdown(GVBD) oocyte($64.2{\pm}7.3nM$). In normal medium after calcium chelator treatment ($10{\mu}M$ BAPTA/AM), the free calcium contents were slightly lower than those in control group. In calcium-free medium, the free calcium content was drastically increased in GVBD($72.7{\pm}3.4nM$) and metaphase I - anaphase I ($88.0{\pm}3.4nM$) oocyte. In maturation rate of oocytes, GVBD rate was high in control group($82.9{\pm}6.55%$) and calcium chelator treatment group($91.2{\pm}4.4%$), but in calcium-free medium group, it was low and then the oocyte was degenerated without polar body formation. Relative content of calmodulin in oocyte was significantly(P<0.001) increased in metaphase I - anaphase I than in GV and GVBD oocyte. The calcium precipitates were observed in mitochondria and cytoplasm of GV oocyte but that were not observed in mitochondria of GVBD and metaphase I - anaphase I oocyte. And then the calcium precipitates reappeared in mitochondria of metaphase II oocyte. The above results indicate that changes in free calcium and calmodulin concentration of oocyte occur according to the maturational stages and the extracellular calcium is required during oocyte maturation. Also change of calcium localization in oocyte occurs according to the maturational stages.

• The Korean Journal of Zoology
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• v.31 no.2
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• pp.81-86
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• 1988

The relationship between cumulus cell expansion, cocyte maturation and metabolic cooperativitiy was investigated by using mouse and pig cocyte-cumulus complexes in vitro. Cocyte germinal vesicle breakdown (GVBD) and cumulus expansion were manipulated with hormones or reagents which increase intracellular cAMP leveL Metabolic cooperativity between oocyte and cumulus cells was assessed by determination of the fraction of radiolabelled uridine marker that was transferred from the cumulus mass to the oocyte. Uptake of uddine marker by mouse and pig cumulus mass was increased by about fourfold of basal level with the stimulation of hormones (human choriononic gonadotrophin, HCG; follicle stimulating hormone, FSH) or cyclic AMP sttmulators (3-isobutyl-1-methylxanthine, IBMX; forskolin) during culture. However, the fraction of uridine that was transferred from the cumulus mass to the cocyte (transfer ratio) was gradually decreased during culture, irrespective with the presence of hormones or stimulators. The decrease of the transfer ratio was not correlated with the state of occyte whether they have GV or not, or with the degree of cumulus expansion. In mouse complexes, HCG induced more significant reducton of transfer ratio than other treatments. These results do not support the idea that modulations of metabolic cooperativity between cumulus cells and oocytes are important for the regulation of meiotic resumption in mammals.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.24 no.3
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• pp.116-122
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• 1998

It is well known that all-trans-retinol is not only very unstable in heat, light, air, and water, but also skin-irritant despite a good anti-wrinkle effect. Therefore, it is very difficult to stabilize retinol and make the safe retinol containing cosmetics by using a certain concentration of retinol with real effect. In order to dissolve these problems and apply retinol for skin care cream, firstly retinol is to be encapsulated in the vesicle called Liposphere (pseudo-liposome) which is made by homogenizing under high pressure the mixtures of lecithin, retinol, caprylic/capric triglyceride, and hydroalcoholic solution ; and then this retinol containing Liposphere is to be intercalated in lamellar liquid crystal layer which is prepared by emulsifying in an optimal ratio the mixtures composed of non-ionic emulsifier (cetearyl glucoside, sorbitan stearate & sucrose cocoate etc), cetearyl alcohol, stearic acid, cholesterol, and ceramide. In addition, the stability of the retinol containing oil in water cream by adding the polymeric emulsifier such as acrylate /C10-30 alkyl alkylate crosspolymer is to be ensured even at 55 C. Retinol containing oil in water cream prepared through above procedure could be very stable at 45 C for at least 50 days. The structure identification of lamellar liquid crystal was determined using polarized light microscope and electron microscope Conclusively, we could make the very stable retinol containing oil in water cream by triple procedure, that is, encapsulation of retinol in Liposphere, intercalation of retinol in lamellar liquid crystal layer, and assurance of the high temperature stability of cream even at 55 C.