• Korean Journal of Poultry Science
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• v.48 no.4
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• pp.185-191
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• 2021

Chicken spermatozoa have the ability to survive in low-temperature environments; however, the effects of low temperature on sperm motility and acrosome damage have not been studied in detail. The present study investigated semen longevity following dilution of rooster semen with Beltsville Poultry Semen Extender (BPSE) and Lake extender in preservation vessels (1.5 mL e-tube and 0.5 mL straw). Spermatozoa motility in the closed-type vessel (0.5 mL straw) was higher than that in the 1.5 mL e-tube on day 3 of preservation (68.6±3.1% vs. 22.1±5.7%). The motility of rooster semen diluted with BPSE in 0.5 mL straw was also higher than that of the Lake extender on day 3 of preservation (57.7±5.6% vs. 37.7±5.4%). Furthermore, acrosome intactness was higher in 0.5 mL straw than in the 1.5 mL e-tube, and the rate of acrosome cap damage increased with preservation days. The present study demonstrates that a closed 0.5-mL straw vessel could be used for low-temperature semen preservation, with an increased motility rate and acrosome integrity in fresh rooster semen.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.4
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• pp.301-310
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• 2009

Objective: To compare the embryonic development and pregnancy results using sperms retrieved from fresh and frozen-thawed testicular tissue in patients with obstructive (OA) and non-obstructive azoospermia (NOA). Methods: A total two hundred twenty-two cycles of TESE-ICSI were performed in OA and NOA. Sperms were retrieved from fresh and frozen-thawed testicular tissue. ICSI was performed patient's own sperm. Fertilization was assessed 16~18 hrs after ICSI. Embryo development and pregnancy rates were analysed. Results: The fertilization rates were significantly different between OA and NOA patients (75.2% vs. 56.7%, p<0.05), however, embryo development did not differ between the groups (96.9% vs. 98.0%). Likewise, OA and NOA groups had no differences in their clinical pregnancy and delivery rates, 33.9% vs. 36.0% and 28.1% vs. 28.0%, respectively. With regard to sperm retrieved from fresh testicular tissue, fertilization rates were significantly different between the OA and NOA groups (76.4% vs. 52.9%, p<0.05); however, embryo development, clinical pregnancy and delivery rates were not different. For sperm retrieved from thawed testicular tissue, the fertilization rates were significantly different between the two groups (74.7% OA group vs. 65.6% NOA group, p<0.05); however, embryo development, clinical pregnancy and delivery rates were not different. Conclusions: Embryo development and clinical pregnancy did not differ in patients with obstructive and non-obstructive azoospermia, whether sperm retrieved from fresh and thawed testicular tissue were used, although the fertilization rates were different. Therefore, ICSI with sperm retrieved from fresh and thawed testicular tissue could achieve relevant clinical pregnancy results in patients with azoospermia.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.1-7
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• 2007

This study was conducted to examine the role of intact cumulus cells during in vitro fertilization (IVF) on sperm penetration, male pronuclear (MPN) formation and subsequent embryo development of oocytes matured and fertilized in vitro. Cumulus-oocyte complexes obtained from the slaughtered gilt ovaries were matured for 44 h in TCM199 containing 10% porcine follicular fluid, epidermal growth factor and hormones. After maturation culture, denuded oocytes or oocytes with intact cumulus cells were coincubated with frozen-thawed boar semen for 8h in a modified tris-buffered medium containing 5mM caffeine and 10mM calcium chloride. Putative zygotes were fixed and examined for sperm penetration and MPN formation (Experiments $1{\sim}3$), or cultured in North Carolina State University-23 medium fo. 156 h (Experiment 3). In Experiment 1, sperm penetration was examined after insemination of denuded oocytes and oocytes with intact cumulus cells at the concentration of $7.5{\times}10^5$ sperm/ml. Optimal sperm concentration for IVF of cumulus-intact oocytes was determined in Experiment 2 by inseminating intact oocytes with $2{\sim}5{\times}10^6$ sperm/ml. In Experiment 3, denuded or intact oocytes were inseminated at the concentrations of $7.5{\times}10^5$ and $4.0{\times}10^6$ sperm/ml, respectively, and in vitro embryo development was compared. Sperm penetration was significantly (p<0.01) decreased in cumulus-intact oocytes compared to denuded oocytes (35.2% vs. 77.4%). Based on the rates of sperm penetration and normal fertilization, the concentration of $4.0{\times}10^6$ sperm/ml was optimal for the IVF of intact oocytes compared to other sperm concentrations. The presence of intact cumulus cells during IVF significantly (p<0.05) improved embryo cleavage (48.8% vs. 58.9%), blastocyst (BL) formation (11.0% vs. 22.8%) and embryo cell number $(22{\pm}2\;vs.\;29{\pm}2\;cells)$ compared to denuded oocytes. In conclusion, these results suggest that intact cumulus cells during IVF inhibit sperm penetration but improve embryo cleavage, BL formation and embryo cell number of porcine embryos produced in vitro.

• Korean Journal of Poultry Science
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• v.16 no.2
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• pp.91-100
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• 1989

This study was conducted to observe 1) the changes of cellular association in seminiferous tubles from 2 to 8 weeks of age, and 2) the cycle phenomena of seminiferous epithelia at 14 weeks of age in Japanese quail. Total 80 birds were examined at a week interval from 2 to 8 weeks, and 14 weeks of age. The results were summarized as follows： 1) The body and testis weights showed most prominent increase during 4 to 5 weeks and 6 to 8 weeks of age respectively. And also the diameters of seminiferous tubles were abruptly enlaged during 6 to 8 weeks of age. 2) Genocytes in the seminiferous tubles were still in existence at 3 weeks of age, however they did not come out after 4 weeks of age. Spermatogonia, primary spermatocytes and spermatids made their first arpearances in the seminiferous from 3, 4 and 6 weeks of age, respectively. Spermatozoa were observed for the first time at 7 weeks of age, but full spermatogenic activity was completed from 8 weeks of age. 3) At 14 weeks of age, the average weight at testis was 3.7g and its ratio to the body weight was approximately 3.0 percent. And at this age, average diameter of seminiferous tubules was 192.08 $\mu\textrm{m}$, and average numbers of spermatogonia, spermatocytes, spermatids and spermatozoa within the cross section of seminiferous tubules were 7.74, 40.81, 28.42, 104.55 and 105.98, respectively. Spermatogonia and spermatid were classfied into 2 and 3 types, respectively. 4) At 14 weeks of age, the cycle of seminiferous epithelium could be divided into S stages with following characteristics. (1) Stage I： Seminiferous tubules showing type I and II spermatids. (2) Stage II： Seminiferous tubules showing type III spermatids only. (3) Stage III： Immature spermatozoa gathered near the sertoli cytoplasm. (4) Stage IV： Forming a bundle of 15-20 spematozoa. (5) Stage V： Spermatozoa bundle leaving the sertoli cytoplasm into lumen of the seminferous tubule. 5) Usually 2-3 stages of the seminiferous epithelium cycle were concurrently appeared within a tubular cross section, and frequency of each stage from I to V within cross section of seminiferous tubules were 11.91%, 27.03%, 27.96%, 19.04% and 17.98%, respectively.

• The Korean Journal of Malacology
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• v.14 no.2
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• pp.91-102
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• 1998

새꼬막은 자웅이체로서 난생이었다. 생식소는 내장낭의 간중장선에서부터 족부까지 외측의 대부분을 싸고 있으며, 난소는 난자형성소낭, 정소는 정자형성소낭으로 구성되어 있었다. 완숙란의 세포질에는 많은 난황과립을 축적하고 있으며, 난막의 외측은 얇은 젤라틴상의 피막으로 싸여 있었다. 정자형성소낭벽에는 정원, 정모, 정세포 및 변태한 정자 순으로, 소낭의 내강을 향하여 충상배열을 하며 성숙 발달되었다. 생식주기는 초기 활성기(11-5월), 후기 활성기(5-7월), 완숙기(6-9월), 부분 방출기(7-9월), 방출 및 비활성기(9-4월)로 구분할 수 있었다. 방란, 방정은 수온이 21$^{\circ}C$ 이상으로 상승하는 7월초순부터 9월까지 지속되며, 산란 최성기는 8월이었다. 비만도지수의 월별 변화는 생식주기와 아주 밀접ㅎ나 관계가 있었다. 난자형성소낭과 정자형성소낭에는 eosin에 강하게 염색되는 과립세포와 간충직들이 나타나는데, 이들은 생식소 및 생식세포형성과 발달에 영양을 공급하는 영양세포로 생각되었다.

• Biomedical Science Letters
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• v.6 no.4
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• pp.237-244
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• 2000

It has been reported that plasma membrane activity of the spermatozoa may be susceptible to be influenced by extracellular osmolality and such membranous changes involve infracellular molecular changes, special regard to the structure of membranous lipids, and the accompanying ion-channel of which are closely related with their fluidity of $Ca^{2+}$ and HCO$^{-}_{3}$. It is of common recognition that a certain kind of sterol acceptor player an important to induce lipid fluctuation of the sperm plasma membrane which have been influenced by BSA administration and came in effect to outflow of cholesterol from the spermatozoa and resulted in changes of ionic fluidity to facilitate adenylyl cyclase, and to induce protein tyrosine phosphorylation by increase of cAMP and activation of PKA. Thus it seems likely that an augmentation of the acrosomal reaction is closely related with protein tyrosine phosphorylation. The following experimental results were obtained in the present study; Under the high osmolality conditions, the spermatozoa motility declined significantly and the structural change of the plasma membrane diminished to confirm that the response degrees to the osmolality depended upon the water transfer volume through the plasma membrane and the changes of cellular volume. Those experimental results suggest that a physiological parameter such as low temperature condition played an important role for presentation of spermatozoa and that inducement of spermatozoa activation for reinforcement of protein tyrosine phosphorylation. On the other hand, it seemed likely that the BSA administration as one of sterol accepters might represent a key role also under the high osmolality condition and their result also suggests that osmolality change, special regard to high osmolality condition may play an important role also in the processes of signal transmission.

• The Korean Journal of Zoology
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• v.34 no.2
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• pp.159-172
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• 1991

It has been investigated what could be the selective marker distinguishing the immature from mature spermatozoa and whether fi -glucuronidase and fi -glucosidase are dependent on androgen in the luminal fluid of the epididymis or not. The contents of hexose, hexosamine and sialic acid in the epithelial cells, luminal fluid and spermatozoa of the epididymis were examined and the patterns of protein bands were compared in each group of the luminal fluid by SDS-PAGE. Lactate dehydrogenase, glucose-6-phosphatase, Na+ -K+ -ATPase and MgNa-ATPase showed higher activities in the cauda than the caput epididymal spermatozoa but only $Mg^2$+-ATPase activity appeared to be changed significantly. When the contents of hexose, hexosamine and sialic acid were analyzed and compared quantitatively, those of hexose were significantly different in the luminal fluid of caput and cauda epididymis, those of hexosamine in the epithelial cells and those of sialic acid in the epithelial cells and luminal fluid. When SDS-PAGE has been performed in each group, the band of MW 33-37 KD which was absent in the luminal fluid of caput epididymis appeared obviously in the luminal fluid of cauda epididymis and ako apeared in the cauda sperm crude membrane fraction. In addition, $\beta$ -glucuronidase and $\beta$ -glucosidase activities and their dependence on androgen were measured and the SDS-PAGE patiems of proteins and/or glycoproteins in the luminal fluid were examined. The activities of these two enzymes in the luminal fluid of the epididymis decreased significantly from the 5th day after castration. When testosterone was injected, the activity of $\beta$ -glucuronidase began to increase significantly from the 5th day following injection and that of $\beta$ -glucosidase from the loth day. On the other hand, the band of about MW 21 KD was newly observed in the lumen of caput epididymis when testosterone was administered.

• 대한생식의학회:학술대회논문집
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• 2002.11a
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• pp.79.1-79.1
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• 2002

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.51-51
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• 2003

대농갱이는 체형이 동자개와 비슷하나 지금까지 인공부화는 물론 산란습성에 관해서도 잘 알려져 있지 않은 미개발 어종으로 양식품종의 다양화 및 경쟁력 있는 새로운 품종 개발을 위하여 인공종묘생산 시험을 시도하였다. 본 연구에 사용한 대농갱이 친어는 금강 중류역에서 서식하고 있는 자연산 친어로 암컷 210마리, 수컷 148마리를 사용하였다. 호르몬주사에 의한 인공 배란효과 조사를 위해 암컷 한 마리당 HCG 1,000 IU 주사군(75마리)과 2,000 IU 주사군(135마리)으로 나누어 배란율을 조사하였으며, 수컷은 주사하지 않고 정소를 적출하여 막자사발에 갈아서 등조액과 혼합하여 사용했다. 수정 방법은 난을 착출하면서 정자 등조액을 동시에 흘리는 방법, 난을 그릇에 받은 후 정자 등조액을 혼합 수정하는 방법, 채란용기에 정자 등조액을 미리 넣고 암컷의 난을 짜 넣어 수정시키는 방법 등 3가지 방법을 사용하였다. 부화된 자어는 초기먹이로 알테미아(3일 급이), 실지렁이(6일 급이), 배합사료 순으로 급이하였으며 수온 25~28$^{\circ}C$에 유지관리하면서 성장도 시험을 실시하였다. 호르몬 주사에 의한 배란효과는 HCG 2,000 IU 주사군(채란율 88.9%)이 1,000 IU 주사군(37.3%)에 비해 월등히 높았으며 난의 성숙상태도 좋았다. 수정방법에 있어서는 채란과 동시에 정자 등조액을 흘려 수정시키는 방법과 채란용기에 미리 정자 등조액을 넣고 암컷의 난을 한 마리씩 짜서 수정한 후 바로 부화지로 이동하여 고르게 뿌려 준 경우가 수정율 및 부화율이 좋았다. 부화수온은 23$\pm$1$^{\circ}C$에서 96시간, 26$\pm$1$^{\circ}C$에서는 72시간만에 부화되었으며 부화여건이 좋으면 단시간에 부화할 수 있도록 하는 것이 수생균 발생을 억제할 수 있어서 효과적이었다. 자치어의 성장도 실험에서는 사육수온 25~28$^{\circ}C$에서 직선식으로 나타낼 수 있어서 평균체장은 Y=0.9943X+0.7867, 평균체중은 Y=0.8800-0.6800로 각각 표시되었다.

• Development and Reproduction
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• v.15 no.2
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• pp.151-158
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• 2011

The comparison of the chemical properties of semen of black porgy Acanthopagrus schlegelii long-term acclimated reared in freshwater (BFW) and seawater (BSW) with sperm activity of salinity and ion composition. The chemical properties of seminal plasma on BFW of the factors that most there was not significant difference in the BSW. However, osmolality in seminal plasma of BFW and BSW was $307.0{\pm}4.6$ and $337.3{\pm}10.1$ mOsm/kg, respectively, where BFW showed significant lower concentration in contrast to BSW. Salinity effect on sperm motility of BFW and BSW in 0 psu solution, no sperm motility was observed, whereas in 10 psu solution, both BFW and BSW sperms showed low motility and short time post sperm activation. However, diluted in 20 and 32 psu solutions, highest motility and long time post sperm activation were observed in BFW and BSW sperm. SAI of BFW and BSW varied in depend on the osmolality regardless of ion kind and it showed the highest value in the similar osmolality of artificial seawater (956 mOsm/kg). Accordingly, even in sperm released from BFW, factors initiating sperm motility are determined by osmolality.