• Journal of Embryo Transfer
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• v.32 no.3
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• pp.105-109
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• 2017

Autophagy is an intracellular degradation and recycling system. Oocyte maturation is dynamic process, in which various proteins should be synthesized and degraded. In our previous study, we reported the loci of autophagosome and dynamics of autophagic activity in porcine oocytes during in vitro maturation. In this study, we verified loci of autophagosome in porcine follicular cumulus-oocyte complex by detection of microtubule-associated protein 1A/1B-light chain 3 (LC3) which is the reliable marker of autophagosome. Porcine ovary including various sizes of follicles was fixed within 1 hour after collection from slaughterhouse. After fixation, immunohistochemistry was conducted on sliced ovary tissue containing various sizes of follicles by using LC3 antibody. As a result, LC3 signal was clearly detected in both cumulus and oocytes of various sizes of follicles. We also found ring shaped signal which represent autophagosome near oocyte membrane. Most of the signals in oocytes were localized nearby cellular membrane while evenly dispersed in cumulus cells. Therefore, this result suggests that autophagy occurs in porcine COCs (cumulus-oocyte complexes) at follicular stage.

• Fisheries and Aquatic Sciences
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• v.21 no.11
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• pp.34.1-34.13
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• 2018

Sexual maturity ($L_{50}$), the length at which 50% of fish in a size class are mature, is a key aspect of domestication of new fish species because it guides the procedure for identification of appropriate broodstock size for artificial spawning. In this study, the $L_{50}$ was determined for 1083 Barbus altianalis samples obtained from Lake Edward and the Upper Victoria Nile. Gonads of freshly killed samples were examined macroscopically and verified with standard histological procedures for the maturation stages that were used to determine $L_{50}$. Oocytes and spermatogenic cell sizes were compared for fish obtained from both water bodies. Results indicated that there were no variations in macro gonad features observed for fish from Lake Edward and Upper Victoria Nile. Similarly, there were no significant differences in oocyte sizes (P > 0.05) between the two populations but significant differences in spermatogenic cell sizes were noted (P < 0.05) except for spermatozoa (P > 0.05). This however did not suggest peculiar differences between the two populations for staging the gonads. Consequently, no staging variations were suggested for both populations in determination of $L_{50}$. Sexual maturity was found in the same class size of fork length (FL) 20-24.9 cm and 35-39.9 cm for males and females from both water bodies, respectively. At this FL, however, males were too small, and for good selection of vigor broodstocks for spawning and conservation purposes, they are better picked from class size of 30-34.9 cm FL and above. These findings were crucial for integration of appropriate breeding size in spawning protocol by farmers and fisheries scientists conserving wild B. altianalis populations.

• Applied Microscopy
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• v.30 no.4
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• pp.367-376
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• 2000

The observation using an electron microscope shows that the maturation of the oocyte of African giant snail, Achatina fulica, proceeds over three stages. The oocyte of stage 1 is a small elliptic cell $(220\times400{\mu}m)$ whose light nucleoplasm contains two nucleoli. In its cytoplasm, a number of mitochondria, rough endoplasmic reticula, and ribosomes are found, while yolk granules are not. The nucleus of the oocyte of stage 2 is relatively large in comparison with the volume of cytoplasm, and contains one nucleolus. In the nuclear envelope comprising inner and outer double membrane, there are found a lot of nuclear pores for materials to pass through. A number of mitochondria, Golgi complex and lipid yolk granules appears in the cytoplasm, and proteinous yolk granules begin to form and mature in the vacuoles of various sizes ($0.8\sim3.0{\mu}m$ in diameter). The oocyte of stage 3 has an enlarged nucleolus. Material transportation through nuclear pore is not found any longer. The cytoplasm in this stage is filled with proteinous and lipid yolk granules. The microvilli are developed around the egg plasma membrane.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.11
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• pp.1477-1486
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• 2009

Female germ cell meiotic divisions are typically asymmetric, giving rise to two daughter cells with different sizes. Spindle movements including spindle migration from the oocyte center to the cortex and spindle rotation from parallel to perpendicular (typically in the mouse) at the cortex are crucial for these asymmetric divisions and therefore are crucial for gamete production. Different regulatory mechanisms for spindle movements have been determined in different species and a wide variety of different molecular components and processes that are involved in spindle movements have also been identified in different species. Here, we review the current state of knowledge as well as our understanding of mechanisms for spindle movements in different systems with focus on three main aspects: microtubules (MT), microfilaments (MF) and molecules associated with cytoskeletal organization as well as molecules that are not directly related to the cytoskeleton. How they might interact or function independently during female meiotic divisions in different species is discussed in detail.

• The Korean Journal of Malacology
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• v.31 no.2
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• pp.113-121
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• 2015

Gametogenesis, mating behaviour and spawning of Octopus ocellatus were investigated by histological study. This species is dioecious, and showed a protandry phenomenon. Ooogenesis (in females) and spermatogenesis (in males) can be classified into 3 stages, respectively. O. ocellatus copulates in one of two ways: a male may leap upon a female, mounting her mantle, or a male may sit near the female and extend the hectocotylized third right arm toward her. Spawning occurred between April and June in females, and between March and May in males of O. ocellatus. The spawning period was once a year and the peak took place between May and June. A number of flatened follicle cells, which were attached to an oocyte, were involved in vitellogenesis in the cytoplasm of the vitellogenic oocyte (maturing oocyte), and formation of chorion membrane (secondary egg membrane) of the ovarian eggs. Fecundity per female closely related to GSI was 294-660 eggs (average, 429 eggs). The diameters of the ovarian eggs surrounded by chorion membrane were approximately in the range of 10.10-2.50 mm. Each ovarian egg laid by a female was connected to an egg string. Each egg string was 1-5.5 cm (average 3.6 cm). The total number of eggs laid by a female of this species ranged 218-314, the egg sizes were independent to the size of female adult. this species has a life mode showing some special reproductive characteristics of an annual semelparity as shown in Octopodidae species because we have never seen a female spawning a second time.

• The Korean Journal of Malacology
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• v.23 no.2
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• pp.209-216
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• 2007

The ovarian structure and ultrastructural changes in germ cells during oogenesis of the spiny top shell, Batillus cornutus, have been investigated by light and electron microscopic observations. Specimens of the spiny top shell were collected from the subtidal zone of Wando, south coast of Korea. Spiny top shell was dioecious. The ovary was located on the surface of the hepatopancreas in the spiral posterior region. The ovary had greenish color in the gonadal mature season. The ovary comprised many ovarian lobules, and the ovarian lobule consisted of connective mesenchymal tissue and epithelial cells. Oogonium had a large nucleus with nucleolus of high electron density. In previtellogenic oocyte, small yolk granules of low electron density were scattered in the cytoplasm. Oocytes in the initial vitellogenic stage were connected with the ovarian lobule by egg stalk. The result of TEM observations showed that initial vitellogenic oocyte contained well-developed Golgi complex, endoplasmic reticula, tubular mitochondria and numerous yolk granules of various electron densities and sizes. The electron density, size and quantity of yolk granules which were distributed in the active vitellogenic oocyte were increased compared to the previous stage. Thickness of egg envelope in the late active vitellogenic oocyte was approximately 4.4 $[\mu}m$. Cytoplasm of ripe oocyte was filled with proteid yolk globules of high electron density and lipid yolk globules of low electron density. In this stage, the thickness of egg envelope was approximately 6.5 ${\mu}m$.

• Development and Reproduction
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• v.5 no.2
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• pp.115-122
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• 2001

Production of a mature oocyte is a complex process that requires the close association between oocyte and follicular cells. The present study was conducted to investigate the difference between oocytes with and without close junctional communication with cumulus cells and the involvement of two connexins(CXs) in the interactions. Follicles at different sizes(small: 200～400 ㎛; large:>450 ㎛) were mechanically isolated from PMSG-primed mouse ovaries, and punctured to get cumulus-oocyte complex(COC). Oocytes were released themselves(denuded), with partially attached(partial), or with tightly attached(intact) cumulus cells. Maturation and fertilization capacity of the COCs were measured. Expression of CX 37 and CX 43 was examined by RT-PCR and in situ hybridization. The ratio between intact COC and denude/partial oocytes was 30％(SI) and 70％(SPD) in small follicles, while 55％(LI) and 45％(LPD) in large follicles, respectively. Maturation and fertilization rates of the released oocytes were similar among SI, LPD, LI groups, but those were significantly lower in SPD oocytes. In oocytes, CX 37 was the major CX and CX 43 was not expressed, whereas in the cumulus cells, CX 43 was the major, and CX 37 was the next. Results of the present study suggest that 1) immature oocytes from small follicles with intact cell-to-cell communication with cumulus have the similar quality to that of the oocytes from larger follicles, 2) gap junction between oocyte and cumulus cells may be the heterotypic channel, and 3) we could not explain the difference in the cell-to-cell communication between intact and partial/denuded COCs through the expression of the two CXs.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.289-299
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• 1993

The polypeptide patterns of cellular and follicular components were analysed by SDS-PAGE and two dimensional(2-D)electrophoresis combined with isoelectric focusing (IEF) to establish protein profiles in each of the components in porcine follicles. Oocyte-cumulus complexes were cultured in M16+FCS+Gn at 39 in an atmosphere of 5% CO$_2$, in air for 35 h. At the end of the culture, the zona-free oocyte, ZP alone and cumulus cells were prepared and analysed either on 10% SDS-PAGE for the protein profile at the first dimensional gel or 2-D protein pattern. The amounts of each samples were determined for the visualization with Coomasie brilliant blue (CBB) or silver staining, thus giving useful information for the identification of specific proteins in the components or appropriate amount of samples for proper visualization. Oocyte showed 25 and 114 kd major protein band. Other minor components were additionally visualized with CBB on the same gel after silver staining procedure. Cumulus cells also showed specific proteins which is not present in the oocytes. The number of cumulus cell was proper to give major bands with CBB and additional minor bands with silver staining. To establish the degree of contamination from the remnant of the corona radiata to the ZP, zonae were differently prepared or analysed by SDS-PAGE.The preparation of the ZP in this study did not showed any contamination judged by the protein profile of the components. Also follicular fluid showed its specific protein profile without any significant differences among the different sizes of follicles. The established protein profile of each follicular component should be helpful for the identification and elimination of contaminated components, i. e., antigen preparation or immunological studies. The results also suggest that the preparation of each components in the study was appropriate and can be used for a further sensitive biochemical analysis in mammalian oocytes and early embryos.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.399-407
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• 2001

The microinjection of retroviral vectors into the perivitelline spaces of MII-stage oocytes increased production of transgenic bovine embryos. However, oocytes have various sizes of perivitelline space, and there is the tendency that the oocyte membranes are damageable by micropipettes during the injection of retroviral vectors into perivitelline spaces or oocytes. Thus, it was not always possible to stably inject retroviral vector into perivitelline spaces of oocytes. Here we used sucrose to minimize the damage of the oocyte membrane. When the oocytes were suspended in 0.5% sucrose, poor quality oocytes showed rough cytoplasmic membranes, while good quality oocytes maintained smooth membranes. However, when the tatters were subjected to in vitro fertilization, no significant differences were observed in cleavage rates (82% of control Vs. 84% of sucrose treated oocytes). The Same trends were obtained from the oocytes fertilized after microinjection of LN$\beta$-EGFP and LNC-hGH genes into the perivitelline spares. The rates of cleavage and blastocyst from microinjection of LN$\beta$-EGFP genes were 81 and 25%, and from microinjection of LNC-hGM genes were 53 and 30%, respectively. The result indicated that microinjected oocytes could develop to the blastocyst stages after in vitro fertilization with no significant difference from control group. Moreover, the integration of hGH-gene (by PCR analysis) was detected in 52% of infected cleaved embryos and the expression of EGFP-gene (under a fluoresrence microscope) was also observed in 34% of infected blastocyst. These results indicated that 0.5% sucrose treatment could be an efficient method not only to select good quality embryos but also to inject retroviral vectors into perivitelline spares without any harm and hence improving developmental rates.

• Journal of Animal Science and Technology
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• v.63 no.1
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• pp.1-15
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• 2021

As a result of intensive breeding, litter size has considerably increased in pig production over the last three decades. This has resulted in an increase in farrowing complications. Prolonged farrowing will shorten the window for suckling colostrum and reduce the chances for high-quality colostrum intake. Studies also agree that increasing litter sizes concomitantly resulted in decreased piglet birth weight and increased within-litter birth weight variations. Birth weight, however, is one of the critical factors affecting the prognosis of colostrum intake, and piglet growth, welfare, and survival. Litters of uneven birth weight distribution will suffer and lead to increased piglet mortality before weaning. The proper management is key to handle the situation. Feeding strategies before farrowing, management routines during parturition (e.g., drying and moving piglets to the udder and cross-fostering) and feeding an energy source to piglets after birth may be beneficial management tools with large litters. Insulin-like growth factor 1 (IGF-1)-driven recovery from energy losses during lactation appears critical for supporting follicle development, the viability of oocytes and embryos, and, eventually, litter uniformity. This paper explores certain management routines for neonatal piglets that can lead to the optimization of their colostrum intake and thereby their survival in large litters. In addition, this paper reviews the evidence concerning nutritional factors, particularly lactation feeding that may reduce the loss of sow body reserves, affecting the growth of the next oocyte generation. In conclusion, decreasing birth weight and compromised immunity are subjects warranting investigation in the search for novel management tools. Furthermore, to increase litter uniformity, more focus should be placed on nutritional factors that affect IGF-1-driven follicle development before ovulation.