• Molecules and Cells
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• 제26권6호
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• pp.558-565
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• 2008

Although the fertilizing ability of spermatozoa is greatly reduced after freezing, complete understanding of alterations induced by cryopreservation has not been elucidated. The present study evaluates the effects of cryopreservation on the $Ca^{2+}$ handling of boar spermatozoa using several sperm activators. Intracellular $Ca^{2+}$ signals from single spermatozoa were measured using confocal $Ca^{2+}$ imaging of unfrozen samples and of other spermatozoa after having been frozen. Elevation of the external $K^{2+}$ concentration elicited a three times larger $Ca^{2+}$ increase in fresh spermatozoa than in cryopreserved spermatozoa. Caffeine elicited $Ca^{2+}$ transients with some oscillations in the fresh spermatozoa, but not in the thawed spermatozoa. Depletion of the $Ca^{2+}$ store with thapsigargin induced a rapid rise in $Ca^{2+}$ in the control but generated a smaller increase of $Ca^{2+}$ after thawing. Exposure to progesterone induced a biphasic rise of the $Ca^{2+}$ level in the fresh spermatozoa only. Sperm viability was reduced by cryopreservation. Resting $Ca^{2+}$ levels in fresh and cryopreserved spermatozoa were similar. Longer incubation (2.5 h) of thawed spermatozoa partly recovered the $Ca^{2+}$ response to the interventions. These results suggest that cryopreservation reduces the responsiveness of spermatozoa to depolarization, modulators of the internal $Ca^{2+}$ store and progesterone in terms of the $Ca^{2+}$ signal, thus providing a possible mechanism for reduced fertility observed in cryopreserved boar spermatozoa.

• Applied Microscopy
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• 제34권3호
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• pp.179-184
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• 2004

야생의 우수리땃쥐, Crocidura lasiura의 부정소 미부의 정상과 비정상 형태의 정자를 광학 및 전자현미경으로 비교 관찰하였다. 정상 정자와 함께 다수의 형태 비정상 정자가 관찰되었다. 이것은 비정상 정자의 존재가 개체의 생식력 없음을 나타내는 절대적 지표가 아님을 의미한다. 하지만 비정상 정자의 존재는 개체의 생식력 없음과 연관되며 특히 두부의 형태의 이상은 생식력 없음과 밀접한 관계가 있다. 건강한 야생의 우수리땃쥐, C. lasiura의 부정소 미부에서 관찰된 정자의 비정상 두부의 형태는 다음과 같은 유형으로 나타났다. 1) 핵질의 응축이 이루어지지 않거나 부분적으로 이루어진 형태 2) 첨체의 분열 혹은 파손이 나타나는 형태 3) 첨체 혹은 원형질막이 부분적 혹은 완전히 접혀진 형태 4) 첨체가 핵과 분리되는 형태 5) 첨체가 불규칙적으로 응축하는 형태 6) 잘못된 위치에서 관찰되는 첨단체

• Journal of Animal Science and Technology
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• 제58권7호
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• pp.25.1-25.6
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• 2016

Background: The current study was designed to investigate the effect of supplementation of Flaxseed (Linumusitatisimum) oil on libido and semen quality of Nilli-Ravi buffalo bulls. Methods: In this study, 12 adult healthy bulls kept at the Semen Production Unit, Qadirabad district Sahiwal, were used. These bulls were divided into three equal groups, A, B and C. Group A was kept as control, while in groups B and C supplementation of feed was provided by using flaxseed oil @125 ml/day and 250 ml/day,respectively for 12 weeks. Two ejaculates per animal were collected at 0 day then 5th, 6th, 7th, 8th, 9th, 10th, 11th and 12th week of treatment. In this way a total 216 samples were taken, and each semen sample was evaluated for color, volume, mass activity, percent motility, sperm cell concentration per ml, percentage of live sperm, and plasma membrane integrity. Libido of bulls was also evaluated before every collection. Results: Analysis of data revealed that these parameters were significantly (P < 0.01) increased in flax oil treated animals as compared to control. Conclusion: It was concluded from the present study that flax seed oil has beneficial effects on reproductive health of buffalo bull.

• Animal Bioscience
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• 제34권5호
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• pp.844-854
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• 2021

Objective: The potential of extra virgin olive oil (EVOO), betaine (BET), and ginger (GIN), as natural antioxidants, in reducing negative effects of heat stress on physiological responses, antioxidant capacity, semen quality and fertility of bucks under heat stress were investigated. Methods: Forty adult Animal Production Research Institute line rabbit bucks were distributed randomly into four experimental treatments of ten rabbits each. The first treatment was fed the commercial pellet diet (CPD) without supplementation and served as a control. The other three treatments were fed CPD supplemented with EVOO (300 mg), BET (1,000 mg), and GIN (200 mg) per kg diet for 3 consecutive months during the summer season. Results: Supplementation of EVOO, BET, or GIN improved (p<0.05) the sexual desire, progressive motility, vitality, intact acrosome and membrane integrity, sperm cell concentration, sperm outputs and fertility. Seminal plasma total proteins, globulin, total antioxidant capacity, glutathione and glutathione S-transferase, and initial fructose increased (p<0.05), while total lipids, aspartate and alanine aminotransferases and malondialdehyde decreased (p<0.05) compared with the control. In comparing the natural antioxidants treatments, GIN evoked the largest improvement. Conclusion: The inclusion of GIN (200 mg/kg diet) appeared to improve the sexual desire, semen quality and oxidative stress of bucks. This may be a beneficial supplement for the management of rabbit bucks used in natural mating or artificial insemination.

• Animal Bioscience
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• 제34권2호
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• pp.198-204
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• 2021

Objective: The aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows. Methods: Twelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination. Results: The findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12). Conclusion: It may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.

• Applied Microscopy
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• 제2권1호
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• pp.7-15
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• 1972

난소상피에서 발생한 난원세포는 성장발육하여 제 1차 난모세포를 형성하며, 배란되어 난형성강 개구부에서 제2차 난모세포는 정자의 침입을 받은 후 2회의 감수분열을 하여 두개의 극체를 형성하고 난전핵과 정전핵이 적합하여 난할을 시작한다. 세포질의 변화 : 난원세포에서는 크기가 서로 비슷하던 미토콘드리아가 난모세포로 성장되어 감에 따라 자기증식을하여 제1차 난모세포에서는 크기가 서로 다르고 훨씬 많은 미토콘드리아가 나타난다. 편평낭상이던 소포체는 난모세포로 성장하여 세포질대사가 활발하여 짐에 따라 포상낭의 소포체로 변하며 배란후 난모세포에서는 세포질내 일부위에 층판상을 이룬다. 골지 장치는 성숙된 난모세포에서 더욱 뚜렷이 나타나며 망상의 사구체형성을 하고 그 내에 몇 개의 골지소포를 만들고 있다. 표층과립은 수정전 제1차 난모세포에서 매우 증가되어 있으나 수정후는 점차파괴되어 소실된다. 염색질은 난원세포에서 밀도가 높고 강공을 이루고 있는 이질염색질과 밀도가 낮은 소량의 진정염색질로 되어 있으며 배란전 난모세포에서는 이질염색질이 감퇴하고 진정염색질이 많아진다. 배란후 난모세포에서는 풍부한 진정염색질을 다시 가진다. 인은 난원세포에서 전자밀도가 높은 타원형이며 배란전 난모세포에서는 소실되었다가 배란후 난모세포에서는 다시 타원형의 인이 관찰된다.

• 한국가축번식학회지
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• 제27권2호
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• pp.153-161
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• 2003

본 연구는 돼지 동결-융해 정자의 배양에 의한 체외수정능력과 glycosidase activity를 평가하기 위하여 chlortetracycfine fluorescence분석법을 이용하여, glycosidase가 체외수정능력과 정자침입에 미치는 영향에 대하여 검토하였다. 또한 돼지난자의 투명대내에서 발견된 당잔기에 대한 정자의 glyco-sidase 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 응해 후 배양하지 않았을 때보다 2시간 배양했을 때 더 높게 나타났다. $\beta$-GlcNAc'ase의 activity 는 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 자를 배양하지 않은 경우보다는 배양된 정자에서 높게 나타났다. 그러나 배양시간에 따른 정자의 존성에 대해 glycosidase의 종류에 따른 유의차는 인정되지 않았다. 한편 투명대내 정자의 접착과 입에 대한 또 다른 실험에서, 서로 다른 glyco-sidase가 첨가된 배양액내에서 수정된 정자가 배양시간이 길어짐에 따라 정자의 침입율은 낮아졌다 $\beta$-GlcNAc'ase; P<0.05). 투명대내의 정자접착정도는 glycosidase의 첨가시에 무첨가시보다 접착정도가 더 높았으며, 가장 높은 접착율은 $\beta$-GlcNAc'ase 첨가시 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.

• Reproductive and Developmental Biology
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• 제40권2호
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• pp.15-22
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• 2016

As a one of unsaturated fatty acid, polyunsaturated fatty acids (PUFAs) have multiple actions: as precursor of prostaglandins (PGs), steroid hormone synthesis and energy production in animal reproduction. PUFAs, which include omega-3 (n-3) and omega-6 (n-6), are derived from the diet and changed by diet, species, breed and season. The plasma membrane of spermatozoa in mammals contain various PUFAs. These composition of PUFAs regulate the membrane fluidity and cause lipid peroxidation via generation of reactive oxygen species (ROS). Induced lipid peroxidation by ROS decreased viability and motility of spermatozoa, and it is reduced by addition of antioxidant and low concentration of PUFAs. Because oocytes of animal have a high lipid components, process of oocyte maturation and embryo development are influenced by PUFAs. In in vitro study, oocyte maturation, embryo development, intracellular cAMP and MAPK activity were increased by treatment of n-3 ${\alpha}$-linolenic acid (ALA) during maturation, whereas n-6 linoleic acid (LA) negatively influenced. Also, inhibition of fatty acid metabolism in oocyte influenced blastocyst formation of cattle. PGs are synthesized from PUFAs and various PUFAs influence PGs via regulation of PG-endoperoxide synthase (PTGS). Steroid hormone synthesis from cholesterol is regulated by expression of steroid acute regulator (StAR) protein and mRNA. Exogenous n-3 and n-6 PUFAs altered sex hormone in animal through stimulate or inhibit StAR activity. Because PUFAs altered PG and steroid hormone synthesis, follicular development was influenced by PUFAs. This effect of unsaturated fatty acid could provide information for improvement of reproductive ability in animals.

• 한국발생생물학회지:발생과생식
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• 제19권4호
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• pp.197-207
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• 2015

This study investigated structure and function of the reproductive system in Aplysia kurodai by means of anatomical, histological, and histochemical observation. Reproductive system of this species is consisted of ovotestis, small hermaphroditic duct, ampulla, accessory genital mass and large hermaphroditic duct. The ovotestis is composed of a large number of follicles, and both oocytes and spermatocytes matured in the same follicle. The small hermaphroditic duct is a single tube and contains a swelling, the ampulla, which functions as a storage organ for endogenous sperm and an oviduct. The accessory genital mass is connected to both the small and large hermaphroditic duct, and consisted of three glands: albumen, membrane (winding) and mucus gland. The albumen gland is consisted of granular cells producing basophilic and neutral mucopolysaccharides. The membrane and mucus gland are consisted of granular cells producing acidophilc and sulfated mucopolysaccharides. The large hermaphroditic duct is a single tubular gonoduct linking the accessory genital mass to the common genital aperture but is consisted of two parallel compartments. Internally, these two compartments are incompletely divided by internal septum or fold, which are called as the red hemiduct and white hemiduct, respectively. The red hemiduct functions as an oviduct and the white hemiduct functions as a copulatory duct. The reproductive system of A. kurodai is externally comprised a single tube, i.e., monaulic type. However, internal structure of duct is incompletely divided into oviduct and copulatory duct, i.e., the oodiaulic type.

• 한국가축번식학회지
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• 제15권1호
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• pp.33-39
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• 1991

본 試驗은 caffeine을 함유한 人工 培養液에서 培養한 牛精巢上體 精子의 形態的 變化를 透過 電子顯微鏡으로 관찰하였다. 電子顯微鏡에서 관찰한 결과 채취 직후의 無培養 精子는 대부분(79.8%)에서 구조상의 變化없이 원형 그대로의 原形質膜을 유지하고 있었으나 5 mM의 caffeine을 함유한 BO培養液에 3시간 培養한 精子는 尖體의 구조상 4가지 形態로 분류되었다. 즉 無反應, 胞狀化, 尖體離脫 및 退行으로서 각각 29.4%, 45.6%, 17.8% 및 7.2%였다. 이러한 결과는 精子膜의 胞狀化가 정상적인 尖體反應이며 尖體離脫 精子는 死滅精子라는 것과 caffeine이 牛精巢上體 精子의 尖體反應을 誘起시킬 수 있다는 것을 示唆하고 있다.