• Korean Journal of Ichthyology
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• v.23 no.2
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• pp.106-110
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• 2011

Fine structure of spermatozoa of the bastard halibut (Paralichthys olivaceus) is described. Its structure is characterized by acrosomeless, having a spherical head region and a small mid piece containing five mitochondria disposed one row. The flagellum inserts centrally on the nucleus and has lateral ribbons. The present study supports the suggestion that spermatozoal ultrastructure is of taxonomic importance and may be a phylogenetic criterion in teleostei.

• Korean Journal of Ichthyology
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• v.21 no.2
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• pp.81-86
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• 2009

The spermiogenesis and mature spermatozoa of Pseudobagrus fulvidraco were described by means of scanning and transmission electron microscopy. Spermiogenesis is characterized by lateral development of the flagellum, nuclear rotation, deep nuclear fossa formation and compaction into thick granules. The spermatozoa exhibit a round head containing a nucleus that lacks an acrosome, and having a midpiece and a flagellum. The midpiece is small and has a short cytoplasm including several mitochondria separated from the tail by the cytoplasmic canal. The flagellum contains the 9+2 classical axoneme structure and has two axonemal fins. The presence of axonemal fins in the flagellum is a common character in Bagridae. The interrelationships among the Bagridae as well as other teleosts are herein discussed.

• The Korean Journal of Zoology
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• v.39 no.2
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• pp.182-189
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• 1996

It has been known that spermatozoa should obtain their fertilizing ability through capacitation and acrosome reaction, and that in these processes of fertilization, Ca2+ platys an important role for their conjugation. Therefore the present study has been designed in order to clarify the effect of fluctuation of the media Ca2+ level and the intracellular concentration of the spermatozoa on the acrosomes. During the incubation of spermatozoa, a considerable fluctuation in the media Ca2+ level has been observed after the BSA administration and the media concentration of Ca2+. It is deduced that these fluctuation rates may have an effect on the acrosome reaction. The fluctuation of K+ flux has been observed in accordance with the incubation period over time, and it's concentration seems to be closely related with the acrosomal reaction. The respiratory exchange rate (RERI of the spermatozoa is kept more regular in the BSA and Cacl2 administration groups than the non-administration group. Based on the experimental findings, it is possible to deduce a hypothesis from these findings that physiological activities of the acrosome reaction are not functionally related to the media Ca2+ level and the intracellular influx of Ca2+ concentration, although Ca2+ platys an important role as a stimulating factor in the acrosome reaction.

• Korean Journal of Ichthyology
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• v.31 no.1
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• pp.1-6
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• 2019

The spermatozoa of Hemiculter eigenmanni is similar to other cyprinid by spherical head containing a nucleus with highly condensed chromatin, a short midpiece with mitochondria and a flagellum located tangentially to the head. The fine structure of cyprinid spermatozoa described classical characteristics of Cyprinidae spermatozoon comprising the absent of acrosome, the shallow nucleal fossa, postnuclear distribution of mitochondria and the lateral insertion of flagellum. However there were some structural differences for their morphology, in the mitochondria and the orientation of centrioles. The proxomal and distal centrioles are oriented approximately $145^{\circ}$ to each other. The mitochondria of 8 or 10 in number are arranged in two or three layers.

• Development and Reproduction
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• v.2 no.2
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• pp.141-148
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• 1998

The fine structure of spermatozoa of Pungtungia herzi was examined with scanning and transmission electron microscopies. The spermatozoa of p. herzi are approximately 37.4 ${\mu}{\textrm}{m}$ in length and a relatively simple cell with a spherical nucleus, a short midpiece and a tail. The acrosome is not present as in most teleost fishes. The ultrastructure of spermatozoa represents typical characteristics of cyprinid spermatozoa including the lateral insertion of flagellum, the organization of centriolar complex in shallow nuclear fossa, and the occurrence and asymmetrical arrangement of mitochondria. In the nuclear envelope and mitochondrion, however there were some morphological differences for their ultrastructure. The nuclear envelope is severely undulated and the shallow nuclear fossa contains two centrioles which are at the angle of some 130$^{\circ}$ each other. The most significant feature can be observed with the mitochondrion; five or more mitochondria, which are shown in primary spermatocyte, fuse to form a single one in the mature spermatozoon. The mitochondrial aspect is different from that of other cyprinid spermatozoa, where their mitochondria have a conventional aspect and never fuse to form a mitochondrial derivative. In terms of sperm evolution the fused mitochondria are regarded as the apomorphic character in comparison with the separate mitochondria. The single mitochondrion is not reported in cyprinid spermatozoon except the case of Rhodeus.

• Applied Microscopy
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• v.33 no.3
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• pp.243-250
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• 2003

Ultrastructural changes of the male germ cells and structure of spermatozoa in Paralichthys olivaceus were examined by means of the light and transmission electron microscopes. The spermatogonium has a large nucleus with a single nucleus with a single nucleolus in the interphase. Primary spermatocytes are identified by the formation of the synaptonemal complex in the karyoplasm. The secondary spermatocytes are more concentrated and contains numerous cell organelle in the cytoplasm. The nucleus of spermatid in spermiogenesis is more condensed in the karyoplasm, and show spherical structure in shape. Mitochondria of the spermatids are observed in the lower portion of the nucleus. The spermatozoon consists of the head, mid piece and tail. The acrosome is not observed in the head. Axial filaments of the flagellum consists of nine pairs of the peripheral microtubules and one pair of the central microtubules.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.411-417
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• 1998

The objective of this study was to dertermine the effects of oviductal fluid and oviductal conditioned medium on polyspermy and in vitro development of porcine oocytes. The addition of oviductal fluid and oviductal conditioned medium in the prefertilization and fertilization medium significantly decreased polyspermy rates and the mean number of spermatozoa in penetrated eggs(P＜0.05). The acrosome reaction rate significantly increased when spematozoa were exposed for 1.5, 3, 4.5h in oviductal fluid and oviductal conditioned medium(P＜0.05). When oocytes cultured for 192h, the percentage of oocytes that developed to the morula and blastocyst stage was higher in culture medium with oviductal fluid and oviductal conditioned medium than without oviductal fluid and oviductal conditioned medium(P＜0.05). These results indicated that the oviductal secreations will effectively reduce both the polyspermy rates and the mean number of spermatwa in penetrated eggs. And the presence of culture with oviductal fluid and oviductal conditioned medium promotes in vitro development of porcine oocytes.

• Journal of Embryo Transfer
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• v.14 no.1
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• pp.69-77
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• 1999

본 연구는 돼지 정액의 동결과정동안 flow cytometric 분석에 의한 정액내 생존정자의 비율을 조사하여 주관적으로 평가되는 활력 및 정상첨체율(normal apical ridge ; NAR)과 비교하여 정자의 손상과 생존성에 대한 적절한 평가법을 찾기 위하여 실시하였다. 동결과정 중 정액채취, 냉각, 예비동결 및 동결융해 후에 flow cytometric 분석에 의한 정자 생존율은 각각 93.0$\pm$3.6, 85.1$\pm$3.9, 28.9$\pm$6.8 및 26.1$\pm$5.9%이었다. 동결처리동안에 생존율은 예비동결 및 동결융해 후 가장 많은 정자사멸로 동결상태 이전의 생존율보다 유의적으로 낮게 나타났다. (p<0.05). 평가기법으로 정액 채취시 활력, NAR율 및 생존율을 조사한 결과 각각 91.0$\pm$4.2, 96.8$\pm$2.5 및 92.2$\pm$3.2%로 NAR율이 생존성 및 활력보다 높게 평가되었으며, 생존율이 활력보다 다소 높게 평가되었다. 그러나 동결융해 후에는 각각 44.0$\pm$8.9, 49.0$\pm$7.9 및 35.6$\pm$9.7%로 활력이 생존율보다 다소 높게 평가되었다. 전체적으로 NAR율은 활력은 생존율보다 높게 평가되었으며, SYBR-14 / PI(propidium iodide) 이중형광염색법에 의한 flow cytometric 평가법으로 생존율은 동결되지 않은 정액에서의 활력 및 NAR 평가보다 다소 민감하게 나타났다. 이러한 결과로 미루어보아 SYBR-14 / PI 형광염색에 의한 flow cytometry의 생존성 평가는 동결되지 않은 정액의 평가방법으로는 적절하지만 동결된 정액의 생존성 평가는 부적절한 것으로 사료되었다.

• Journal of Embryo Transfer
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• v.26 no.3
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• pp.187-193
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• 2011

This study was carried out to establish most suitable freezing condition, to evaluate the different glycerol concentration of freezing and thawing rates on motility, viability, membrane integrity and acrosome intecrity of frozen Korean Jeju Black Bull spermatozoa, Semen was collected from a Korean Jeju Black Bull using an artificial vagina and transported to the laboratory. The semen was extended gradually 1:5 then cooled slowly for 2 hrs to 4$^{\circ}C$. The semen was diluted 1:1 with cryoprotectant extenders (3%, 5% and 7% glycerol) and equilibrated for 2 hrs at cold chamber and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 minutes, above 5 cm for 10 min and above 8 cm for 10 min. And then the frozen straw was plunged into LN$_2$. The presented straws were examined the viability and motility after thawed at 37$^{\circ}C$ water bath. The viability and membrane integrity immediately post-thawing were significantly higher in samples frozen in 7% glycerol than 3% and 5% glycerol (p<0.05). After CTC staining to assess acrosome integrity, F pattern was significantly increased, but B pattern was significantly decreased in 7% glycerol (p<0.05). Freezing distance of 5 cm from liquid nitrogen and pre-cooling for 10 min yield better survival and membrane integrity, but not significant difference. However, AR pattern according to CTC staining was significantly decreased in 3 cm for 5 min.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.25-25
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• 2003

본 연구는 돼지 동결-융해 정자의 배양에 의한 체외수정능력과 glycosidase activity의 관계를 검토하였으며, 또한 돼지난자의 투명대내에서 발견된 당잔기에 대한 정자의 glycosidase 특이성을 확인하기 위하여 $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase 및 N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase)의 activity를 분석하였다. 그 결과 glycosidase activity는 동결정자의 융해 후 배양하지 않았을 때보다 2시간 배양했을 때 더 높게 나타났다. $\beta$-GlcNAc'ase의 activity는 정자 배양 유무에 관계없이 다른 glycosidase 처리시보다 최소한 2배 이상 높게 나타났다. 또한 첨체반응이 유기된 정자의 비율은 glycosidase ($\alpha$-D-mannosidase; P<0.05)에 의해 영향을 받았으며 정자를 배양하지 않은 경우보다는 배양된 정자에서 높게 나타났다. 그러나 배양시간에 따른 정자의 생존성에 대해 glycosidase의 종류에 따른 유의차는 인정되지 않았다. 한편 투명대내 정자의 접착과 침입에 대한 또 다른 실험에서, 서로 다른 glycosidase가 첨가된 배양액내에서 수정된 정자가 배양시간이 길어짐에 따라 정자의 침입율은 낮아졌다($\beta$-GlcNAc'ase; P<0.05). 투명대내의 정자접착 정도는 glycosidase의 첨가시에 무첨가시보다 접착정도가 더 높았으며, 가장 높은 접착율은 $\beta$-GlcNAc'ase첨가시 나타났다. 또한 모든 glycosidase 처리시 2시간 배양한 정자보다는 배양하지 않은 정자에서 투명대에 대한 접착정도가 높게 나타났으며, $\alpha$-D-mannosidase의 처리시 유의적인 차이를 보였다(P<0.05). 본 연구의 결과, $\beta$-GlcNAc'ase가 주로 돼지정자의 원형질막내에 존재하는 것으로 추측되며, 배양된 정자에 의한 투명대 접착정도와 침입율이 낮았음에도 불구하고 glycosidase activity가 증가하는 것으로 나타났다