• Advances in Traditional Medicine
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• 제1권1호
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• pp.8-27
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• 2000

Components of extracellular matrix and the matrix-degrading enzymes are some of the key regulators of tumor metastasis and angiogenesis. Hyaluronic acid (HA), a matrix glycosaminoglycan, is known to promote tumor adhesion and migration, and its small fragments are angiogenic. Until now, we have compared levels of hyaluronidase, an enzyme that degrade HA, in normal adult prostate, benign prostate hyperplasia and prostate cancer tissues and in conditioned media from epithelial explant cultures, using a substrate (HA)-gel assay and ELISA-like assay (Kim et al., unpublished results). The present review described an overall characterization of hyaluronidases and its application to human diseases. The hyaluronidases are a family of enzymes that have, until recently, deed thorough explication. The substrate for these enzymes, hyaluronan, is becoming increasingly important, recognized now as a major participant in basic processes such as cell motility, wound healing, embryogenesis, and implicated in cancer progression. And in those lower life forms that torment human beings, hyaluronidase is associated with mechanisms of entry and spread, e.g. as a virulence factor for bacteria, for tissue dissection in gas gangrene, as a means of treponema spread in syphilis, and for penetration of skin and gut by nematode parasites. Hyaluronidase also comprises a component of the venom of a wide variety of organisms, including bees, wasps, hornets, spiders, scorpions, sh, snakes and lizards. Of particular interest is the homology between some of these venom hyaluronidases and the enzyme found in the plasma membrane of mammalian spermatozoa, attesting to the ancient nature of the conserved sequence, a 36% identity in a 300 amino acid stretch of the enzyme protein. Clearly, hyaluronidase is of biological interest, being involved in the pathophysiology of so many important' human disorders. Greater effort should be made in studying this family of enzymes that have, until recently, been overlooked. Also, oriental medical application of the hyaluronidase will be discussed with respect to inhibition and suppression of inflammation and malignacy.

• 한국발생생물학회지:발생과생식
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• 제2권1호
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• pp.101-109
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• 1998

1993년 1월부터 12월까지 한국 서해 군산시 오식도 조간대에서 채집한 대맛조개를 대상으로 정자형성과정 및 생식주기를 조직학적 및 투과형전자현미경으로 조사하엿다. 대맛조개(Solen grandis)는 자웅이체이다. 본 종의 완숙정자의 형태 구조는 다른 이매패의 정자들이 갖는 원시형(primitive type)으로 작은 두부와 한 개의 두모첨체를 가지며, 편모축사를 둘러싸고 있는 4개의 미토콘드리아로 이루어진 짧은 중편을 갖는 것이 관찰되었다. 완숙정자 두부의 길이는 대략 2 \mu m정도였고, 정자 미부의 길이는 약 20 \mu m 정도였다. 정자 미부 편모의 axoneme은 중앙의 2개의 미세소관(microtubule)과 주변에 위치한 9쌍의 미세소관으로 구성되어 있었다. 본 종의 방정기는 6월과 7월 사이로써 주된 방정시기는 해수수온이 20 \circ C 이상 상승하는 7월에 일어났다. 생식주기는 초기활성기 (12-1월), 후기 활성기 (1-3월), 완숙기 (3-8월), 부분방정기 (6-7월), 퇴화 및 비활성기 (8-12월)의 연속적인 5단계로 구분할 수 있었다.

• 한국발생생물학회지:발생과생식
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• 제2권1호
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• pp.63-68
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• 1998

본 연구에서는 보다 효율적인 동결 보존법을 수립하기 위하여 현재 사용되는 동결 보존액과 동결방법을 정자의 운동성 측면에서 비교해 보았다. 즉, 세 종류의 조성이 다른 동결보존액인 TYB, dithiothreitol을 첨가한 TYB+DTT, KS II 등이 동결보존 전후에 있어 운동성에 미치는 영향을 조사하였으며, 또한 vapor freezing 방법과 computerized freezer를 사용한 동결방법이 정자 운동성에 미치는 영향을 알아보았다. 정자의 분석은 현미경적 방법과 두 종류의 컴퓨터 정자 자동측정기인 SAIS와 Hamilton Thorn을 사용하여 동결 전 후의 정자 운동성과 VCL, VSL, VAP, ALH, LIN 등의 sub-motility 패턴을 측정하였다. 정액성상이 정상인 군에서 동결보존액을 비교한 실험결과는 TYB군과 TYB+DTT군, 그리고 KS II군의 융해 후 운동성이 각각 28.3%, 23.0%, 34.8%로 KS II군이 우수하였고, 동결방법을 비교한 실험에서는 vapor freezing 군과 computerized freezing 군의 융해 후 정자 운동성이 각각 27.8%, 33.2%로 유의차는 없었다. 또한 무력정자증을 보인 정액군에서는 TYB군과 TYB+DTT군, 그리고 KS II군에서 융해후 정자 운동성이 각각 13.6%, 10.0%, 18.5%로 여기 KS II군이 우수하였으며, vapor freezing군과 computerized freezing군의 융해 후 정자 운동성은 12.8%, 12.9%로 유의차가 없었다. 이상의 결과로 보아 운동성이 정상인 정액군과 무력정자증을 보이는 정액군에서 KS II를 사용해 동결하는 것이 TYB나 TYB+DTT를 사용하는 것보다 운동성 있는 정자를 회수하는데 더 효율적이며, 동결방법 측면에서는 vapor freezing 방법과 computerized freezing방법간에 큰 차이가 없음을 볼 수 있었다.

• 한국수정란이식학회지
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• 제10권2호
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• pp.121-129
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• 1995

To examine the critical effect of oxygen concentration on embryonic development, in vitro fertilized embryos were cultured in media(TCM199 vs. SOF) supplemented sera(1O% FCS vs. 10% HS) with and without bovine oviduct epithelial cells under two gas atmosphere (5% $CO_2$ in air vs. 5% $CO_2$, 5% $O_2$, 90% $N_2$). Oocytes, obtained from abattoir ovaries, were matured in EGF containing TCM199 medium co-cultured with BOEC for 24 hours, followed by exposure to frozen-thawed, heparin4reated spermatozoa in TALP for 30 hours. And then early embryos(1~2 cell) were cultured in both TCM199 and SOF supplemented with 10% FCS or 10% RS under 5% $CO_2$ in air or 5% COi, 5% $O_2$, 90% $N_2$. Development to morulae and blastocysts was recorded on days 7, after the start of in vitro fertilization. The developmental rates of in vitro fertilized embryos to morulae and blastocysts cultured in SOF with BOEC under 5% $CO_2$, 5% $O_2$, 90% $N_2$(24.4%) were significantly(p<0.05) higher than cultured in SOF with BOEC under 5% $CO_2$ in air(14.1%) at seven days after in vitro fertilization. When early bovine embryos were cultured in TCM 199 and SOF under two different gas atmosphere, there were no significant differences in the developmental rates to morulae and blastocysts between supplements of 10% FCS and 10% HS. The rates of development to morulae and blastocysts were significantly(p<0.01) higher in TCM 199 with BOEC(24.7%) than TCM199 without BOEC(10.9%) under 5% $CO_2$ in air, otherwise SOF without BOEC(36.4%) were significantly (p<0.05) higher than in SOF with BOEC (24.4%) under 5% $CO_2$, 5% $O_2$, 90% $N_2$. In summary, these experiments have proved that the culture system in SOF supplemented 10% ES is effective on in vitro development of early bovine embryos under 5% $CO_2$, 5% $O_2$, 90% $N_2$. In addition, it is effective to development of bovine embryos that TCM 199 should be co-cultured with BOEC and SOF should be cultured without somatic cells under two different gas atmosphere.

• Asian-Australasian Journal of Animal Sciences
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• 제22권4호
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• pp.507-515
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• 2009

A series of experiments were conducted to determine the suitable freezing and thawing temperatures for the freezing of boar semen in 5 ml maxi-straws. The ultrastructure, in vitro fertilization (IVF) and artificial insemination (AI) of frozen-thawed semen were also be evaluated. The 5 cm freezing height gave the best results not only in post-thaw motility rate (54.00%), but also in normal acrosome morphology rate (NAR) (80.23%). There was no significant difference in the post-thaw motility between different thawing temperatures and corresponding thawing times (p>0.05); the group of $52^{\circ}C$ and 25 s gave the highest motility rate (45.00%). As a whole, not only from the motility but also the NAR, thawing at $42^{\circ}C$ was better than the other two treatments. In the freezing packages, 5 ml maxi-straw gave a little lower mobility (40%), viability rate (49.58%), plasma membrane integrity rate (53.91%) and NAR (52.65%) than the 0.25 ml straw, but there was no significant difference between the two straw volumes (p>0.05). The IVF capacity of frozen-thawed semen in this experiment was similar to fresh semen. From ultrastructure observation, the main damage to boar spermatozoa after freezing was seen in the acrosome, such as swelling and formation of vesicles. After AI in recipient Shanghai White sows, frozen-thawed semen from 5 ml maxi-straws and pellets produced 72.2% and 80% conception rate and 7.8 and 8 litter sizes, respectively, and there was no significant difference between the 5 ml maxi-straw and the pellet (p>0.05).

• Asian-Australasian Journal of Animal Sciences
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• 제14권8호
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• pp.1188-1195
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• 2001

A tetraparental chimeric bull was successfully produced by aggregating bovine IVF embryos of F1 (female Holstein${\times}$male Japanese Black) and F1(female Japanese Brown${\times}$male Limousin) and culturing in vitro without the zona pellucida at Yamaguchi Research Station in Japan. In the microsatellite genotyping, 12% (28/228) microsatellite primer sets ware potentially useful for this parentage analysis in the chimeric bull, 78.6% (22/28) of microsatellite present in the chimeric bull were uniquely contributed from the Japanese Black and 21.4% (6/28) from Limousin. This chimeric bull semen was used in producing IVF embryos. The chromosome preparations were made from peripheral lymphocytes. Based on chromosome analysis the Chimera had apparently normal chromosomes (29 acrocentric pairs, one large sub metacentric X chromosome and one small sub metacentric Y chromosome). The proportion of acrosome reacted spermatozoa after 1 h of incubation was higher (p<0.01) with the Chimera than with the Holstein and in Japanese Brown bulls. But did not differ from Japanese Black and Limousin bull sperm. Fertilization rates observed after 5 h of sperm-oocyte incubation with Chimera sperm were higher (p<0.05) than with Japanese Brown and (p<0.01) than with Holstein sperm, but did not differ from Japanese Black and Limousin sperm. The cleavage rates of IVF oocytes inseminated with Chimera sperm were also higher (p<0.001) compared with Holstein, (p<0.01) Japanese Brown and (p<0.05) Limousin, but did not differ from Japanese Black sperm. The blastocyst rates of IVM oocytes inseminated with sperm were higher (p<0.05) than in Limousin, Japanese Brown and Holstein, but did not differ from Japanese Black. Chimeric cattles were produced by aggregation of parthenogenetic (Japanese Brown) and in vitro fertilized (Holstein) bovine embryos at the Yamaguchi Research Station in Japan and by aggregation of parthenogenetic (Red Angus) and in vitro fertilized (Holstein) embryos at the St. Gabriel Research Station in Louisiana. The aggregation rate of the reconstructed demi-embryos cultured in vitro without agar embedding was significantly lower than with agar embedding. The aggregation was also lower when the aggregation resulted from a whole parthenogenetic and IVF-derieved embryos cultured without agar than when cultured with agar. The development rate to blastocysts, however, was not different among the treatment. To verify parthenogenetic and the cells derieved from the male IVF embryos in blastocyst formation, 51 embryos were karyotyped, resulting in 27 embryos having both XX and XY chromosome plates in the same sample, 14 embryos with XY and 10 embryos with XX. The viability and the percentage of zonafree chimeric embryos at 24 h following cryopreservation in EG plus T with 10% PVP were significantly greater than those cryopreserved without PVP. Pregnancies were diagnosed in both stations after the transfer of chimeric blastocysts. Twin male and single chimeric calves were delivered at the Yamaguchi station, with each having both XX and XY chromosomes detected. Three pregnancies resulted from the transfer of 40 chimeric embryos at the Louisiana station. Two pregnancies were Jost prior to 4 months and one phenotypically chimeric viable male born.

• 한국수산과학회지
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• 제22권5호
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• pp.342-350
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• 1989

낙동강 하구지역에 서식하는 자주새우를 대상으로 1988년 6월부터 1989년 5월까지 성성숙과 산란에 대하여 조사한 결과는 다음과 같다. 1. 난소는 두흉부의 심장 하방에 위치하며 심장을 중심으로 전방은 촉각동맥 및 안동맥을 둘러싸고 둥글게 융합되어 있고, 후방은 1쌍의 관상으로 복부 2마디까지 뻗혀있다. 정소는 심장직하에 위치하며 심장을 중심으로 전방은 관상으로 곡정관의 형태를 취하고 있으며 후방은 1쌍의 관상의 형태를 띠고 있고, 이곳으로부터 연결된 저정낭이 제5흉지에 개구되어 있다. 2. 난자 및 정자의 배우자형성은 계절과 관계없이 단기간에 반복되고 있으며, 생식소상피의 전 구역에서 동시에 이루어지고 있다. 완숙난의 장경은 $430{\mu}m$ 전후이고, 변태된 정자는 두부 핵질이 뚜렷한 타원형의 구조를 나타낸다. 3. 생식주기는 약 40일을 간격으로 단주기성이며, 11월을 전후한 동계를 제외하고는 주년 산란종이다. 4. 포란된 난의 부화기간은 수온 $21^{\circ}C\~24^{\circ}C$의 실내사육에서 $12\~14$일이 소요되었다. 5. 포란된 개체의 평균 부화유생수는 1,602마리였고, 전장 32mm에서 1,372마리, 38.2개개에서 1,650마리였다.

• 한국수산과학회지
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• 제38권5호
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• pp.309-315
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• 2005

Changes of sex steroid hormones in the plasma of yellowfin goby, Acanthogobius flavimanus were investigated in relation to the gonadosomatic index (GSI), the hepatosomatic index (HSI) and gonadal development. The GSI in females rose rapidly in November and remained high from December to May $(7.26\pm0.89­6.62\pm0.02)$. The Male's GSI also increased gradually from November and was highest in May $(0.16\pm0.08)$. The HSI in both sexes was in reverse correlation with the GSI, and the HSI was low during the spawning season (February-May). In females, the $estradiol-17{\beta}\;(E_2)$ level increased during vitellogenesis (November and December) and reached its maximum $(8.13\pm2.87 ng/mL)$ at the maturing period, in January. $17{\alpha},\;20{\beta}$-dihydroxy-4-pregnen-3-one$(17{\alpha}20{\beta}OHP)$ gradually increased from October $(0.063{\pm}0.02ng/mL)$ to March $(0.16{\pm}0.02ng/mL)$ and increased rapidly in May. The level of testosterone (T) showed a similar tendency of $E_2$. In males, T increased gradually during spermatogenesis from September to December $(0.14{\pm}0.06­0.26{\pm}0.10ng/mL)$ and peaked in January $(0.36{\pm}0.29 ng/mL)$ when the spermatozoa filled the testis. 11-KT also rose markedly in January and then decreased. On the other hand, $17{\alpha}29{\beta}OHP$ in males did not show any clear tendencies.

• 한국산학기술학회논문지
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• 제21권4호
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• pp.490-496
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• 2020

가축유전자원으로서 동결정액을 생산하는 가장 쉬운 방법은 스티로폼박스를 이용한 간이동결법으로 알려져 있다. 본 연구에서는 스티로폼 동결박스를 제작하여 가축의 동결정액 생산에 활용하는 방법을 검토하였으며 칡소 동결정액 생산을 최적화 할 수 있는 방법을 제시하고자 박스의 크기, 액체질소와 노출된 정액 스트로와 거리, 노출시간 및 생산량을 검토하였다. 2가지 동결박스를 비교하여 자료를 확보하였으며 내부 크기는 세로×가로×높이가 23.5×30.5×22.5 cm와 25.5×46.5×26.5 cm로 측정되었다. 액체질소를 5cm 높이로 채우고 액체질소 위 2, 5 및 8cm 높이에서 동결하여 융해 후 생존성을 조사하였다. 칡소 정액을 동결할 경우, 액체질소와의 노출시간은 모두 10분이 적절하였으며 25.5×46.5×26.5 cm 크기의 상자가 높은 생존율을 보여주었다(60.4±5.3% 대 67.2±3.1%). 동결 상자의 최적화 공간은 정자 동결에 가장 중요한 요소로 판단되며 1회 동결 시 최대 생산 가능한 칡소 동결정액은 60개 이상으로 증가시킬 수 있었다. 이러한 정보를 활용하면, 축종에 따라 동결 정액 생산량 결정하고 목적에 맞는 용기를 활용하여 효율적인 동결정액 생산이 가능할 것으로 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• 제15권2호
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• pp.172-178
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• 2002

The present study examined whether treatment of in vitro matured pig oocytes with calcium ionophore (A23187) could prevent polyspermic penetration in vitro. When oocytes cultured for maturation for 33, 36 or 44 h were subsequently treated with $50{\mu}M$ A23187 in medium with fetal calf serum (FCS) for 1, 2 and 3 h and then cultured for 12 h without spermatozoa, virtually no activation occurred. In the absence of FCS, however, 31-42, 45-49 and 56-64% of oocytes were activated, respectively. When oocytes treated with $50 {\mu}M$ A23187 in medium with FCS for 3 h were inseminated in vitro, the penetration rates (14-57%) were lower (p<0.01) with a higher (p<0.01) incidence (35-67%) of monospermy compared with untreated oocytes (69-80% penetration and 15-17% monospermy). However, sperm penetration was completely blocked in all oocytes treated with A23187 in the absence of FCS. When oocytes matured for 33 h were treated with different concentrations of A23187 for 3 h and inseminated in vitro, the penetration rate did not change but there was an increased incidence (p<0.05) of monospermy at $10-20{\mu}M$ and $2.5-5{\mu}M$ A23187 in the presence and absence of FCS, respectively, compared with at $0{\mu}M$ A23187. With these lower concentrations of A23187, treatment of oocytes for at least 60 and 30 min in the presence and absence of FCS, respectively, was required to increase the incidence of monospermy without reducing penetration rate. These results indicate that a high concentration ($50{\mu}M$) of A23187 in medium without FCS, but not in medium with FCS, stimulated in vitro matured pig oocytes to induce parthenogenetic activation and a complete block to sperm penetration in vitro. However, treatment of oocytes with lower concentrations of A23187 ( $10-20{\mu}M$ and $2.5-5{\mu}M$) both in the presence and absence of FCS maintained sperm penetration in vitro and increased the incidence of monospermy.