• Journal of Aquaculture
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• v.20 no.3
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• pp.173-177
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• 2007

An experiment was performed to obtain cryopreservation techniques of starry flounder (Platichthys stellatus) sperm. Milt obtained from 24 males were cryopreserved using two diluents, artificial seminal plasma (ASP) and Stein's solution (SS) with three cryoprotectants, dimethyl sulfoxide (DMSO), methanol, and glycerol concentrated from 5% to 20%. Post-thaw sperm activity (motility and/or speed) revealed the highest in 10% DMSO and 15% methanol in ASP and SS as diluent. Motility and speed of cryopreserved sperm were decreased according to increase of glycerol concentration. To conclude, DMSO was a better cryoprotectant than methanol or glycerol for cryopreservation of starry flounder sperm. Glycerol was incongruent cryoprotectant because of toxic to starry flounder sperm. Most cryopreserved spermatozoa without cryoprotectant showed the enlarged head with granulated chromatin and ruptured plasma membrane by freezing and thawing injuries compared with unfrozen normal spermatozoa.

• Applied Microscopy
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• v.31 no.3
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• pp.245-256
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• 2001

The morphological characteristics of spermatogenic cells during the spermiogenesis of Paradoxornis webbiana were studied by transmission electron microscope. Spermiogenesis of P. webbiana was divided into ten phase. The chromatin granules became fibrous granules at the Golgi phase, gradually condensed at the cap phases, condensed as a stick at the acrosomal phase, and finally, a perfect nucleus was formed at the maturation phase. The formation of sperm tail began at the early Golgi phase, and completed at the late maturation phase. In particular, the dense materials existed in the sperm neck, which is wedged between the tip of segmented columns and the first mitochondria of the middle piece. The axone in the neck were surrounded by the dense materials. The axonema in spermatozoon contains a 9+2 arrangement of microtubules: 9 doublets, and 2 central single microtubules. Mitochondrial bundles of middle piece were composed of a pair of arms, which surrounded the axone of the middle piece by the $15^{\circ}$ angled-helical structure. The outer membrane of mitochondria were surrounded by microtubules in plasma membrane of the sperm. The undulating membrane had a helical structure, and the sperm plasma membrane was surrounded by undulating membrane.

• Journal of Embryo Transfer
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• v.10 no.2
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• pp.153-161
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• 1995

The present study was performed to investigate the effects of caffeine and heparin on capacitation and acrosome reaction of bovine spermatozoa, effects of antisperm antibodies on acrosome reaction of bovine spermatozoa. The rates of acrosome reaction in control group, caffeine treated group, heparin treated group, caffeine-heparin complex treated group were 40.3, 54.3, 63.3, 72.3%, respectively and there were significant differences among the groups(p<0.01), especially higher in caffeine-heparin complex treated group than the others. The rates of acrosome reaction of antisperm antibodies serum supplemented groups(5, 10 and 20%) were 60.4, 48.9 and 37.1%, respectively and there were significant differences among the groups(p<0.0l), and the more increases in serum concentrations, the more decreases in acrosome reaction, but this phenomenon was not seen in fetal calf serum supplemented group and heifer serum group. When the serum concentration was 5%, the rates of acrosome reactions were significantly lower in fetal calf serum supplemented group than heifer serum group and in antisperm antibodies serum group(p<0.01), and there were no significant differences between heifer serum group and antisperm antibodies serum group(p<0.01). When the serum conecntrations were 10%, 20%, the rates of acrosome reactions were significantly lower in antisperm antibodies serum supplemented group than in fetal calf serum group and in geifer serum group(p<0.01), and there were no significant differences between fetal calf serum group and heifer serum group(p<0.01). These results indicate that caffeine-heparin complex treatment is very effective for inducing acrosome reaction of bovine spermatozoa and that antisperm antibodies block acrosome reaction.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.730-736
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• 1999

The roundnose flounder's (Eopsetta grigorjewi) spermiogenesis and fine structure of the spermatozoon were examined by means of the scanning and transmission electron microscopy, During the spermiogenesis, the chromatin of the spermatid became fine granular form, and progressively condensed into many large globules, finally homogeneously condensed in the spermatozoan head. The main characteristics of the spermiogenesis were the disappearanre of Golgi complex, the appearance of microfilament the reduction of mitochondria and the appearance of Iysosome in the cytoplasm. A spermatozoon consisted of head and tail, but the acrosome was absent. The cytoplasmic collar containing seven mitochondria was observed in the posterior part of the head. The well-developed axonemal lateral fins were observed in the tail. The cross section of the axial filament showed '9+2' axonemal structure of microtubules, and the numerous vesicles were observed in the cytoplasm.

• Applied Microscopy
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• v.33 no.4
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• pp.267-274
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• 2003

The ultrastructures of spermatogenesis and sperm in Xiphophorus maculatus, ovoviviparous fish were investigated by electronmicroscopy The testis of Xiphophorus maculatus contained numerous testicular sacs, and spermatogenesis was synchronized in these testicular sac. In the case of spermatogonium, the nucleus was comparatively large ellipsoidal, and the nucleolus and mitochondria showed a marked development. The size of primary spermatocyte was smaller than that of spermatogonia, and that of secondary spermatocyte was smaller than that of primary spermatocyte. The chromatin of spermatocyte was highly condensed according to their development. The nucleus with electron-dense was round shape. In spermiogenesis, flagella started to be formed and chromatin was more condensed. The mitochondria were rearranged along the tail. The sperm was formed by loss of cytoplasm. The head of mature sperm was long cone shape and had not acrosome. The microtubules of flagella were arranged 9+2 structure. Also, the sperm has a loop-like structure at the end of a tail.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.08a
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• pp.36-37
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• 2001

큰가리비는 자웅이체이다. 난환형성과정은 난모세포의 발달정도에 다라 다르게 나타나고 있다. 난자형성과정은 난원세포기, 전난황형성난모세포기, 초기난황형성난모세포기, 후기난황형성난모세포기, 성숙난모세포기의 연속적인 5단계의 과정으로 나눌 수 있었다. 전난황형성기 난모세포질 내에서는 핵주변 구역에 골지장치와 수많은 공포들 및 미토콘드리아들이 출현하고 있는데 이들은 차후, 지방적 형성에 관여한다. 난황형성전기난모세포(previtellogenic oocyte)에서는 지방적 및 지질과립들이 핵막 근처에서 출현하여 피질층쪽으로 분산되는 반면, 같은 발달 단계의 난모세포질의 피질구역에서는 피질과립들(단백질성 난황과립)이 처음으로 생성되어 난황막 근처의 피질층에서 핵주변 구역쪽으로 분산.분포된다. 난황형성후기 난모세포에서는 세포질 내의 골지장치, 공포, 미토콘드리아, 그리고 조면소포체들이 자율합성에 의해 난황과립 형성에 관여하고 있다. 반면 외인성 물질들인 지질형태의 과립들, 다량의 글리코겐 입자들이 생식상피 내에서 출현하고 있는데. 이들 물질이 생식상피에서 난황막 구조물인 미세융모를 통해 난황형성 후기 난모세포의 날질 내로 통과해 들어가는 현상이 관찰되었다. 이와같은 현상은 난황형성이 일어날 때에 heterosynthesis가 일어나고 있음을 시사한다. 완숙난모세포의 난경은 약 50~60$\mu\textrm{m}$이다. 정자형성과정은 정원세포기, 제1차정모세포기, 제2차정모세포기, 정세포기, 정자기의 연속적인 5단계로 나눌수 있었다. 정셍포기에서 정자로 변태되는 과정 중에 침체의 분화과정이 있는데 이에는 1. Golgi phase, 2. Cap phase 3. acrosome phase, 4. maturation phase의 단계를 거쳐 첨체가 완성된다. 정자는 원시적 형태를 이루고 있으며 4개의 미토콘드리아가 부핵을 형성하고 있다. 완숙정자 두부의 길이는 대략 $3 \mu$m 이며, 미부의 길이는 약 $30 \mu$m정도이다. 정자 미부편모의 axoneme은 중앙의 2개의 미세소관(microtubule)과 주변에 위치한 9개의 2중 미세소구관(microtublue)으로 이루어져 있다.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.239-246
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• 1992

Capacitated and acrosome~reacted spermatozoa were microinjected into the perivitelline space of bovine oocytes matured in vitro. Oocytes obtained from the ovaries of slaughtered heifers and cows were cultured in vitro in the TCM-199 supplemented with 20% FCS for 24 hr at 39$^{\circ}C$ under an atmosphere of 5% CO$_2$ 8% O$_2$. Fresh or frozen spermatozoa were incubated for 2 hr at 39°C under an atmos-phere of 5% CO$_2$, 8% O$_2$ in Ham's F-lO medium containing 0.75% BSA for capacitation, and kept for 30 min in culture medium containing 12 mM of dbcGMP and lOmM of immidazol for acrosome resction. One motile spermatozoon was injected into the perivitelline space of each oocyte. The 2nd polar body and the pronuclei were observed in 9.5% and 5.4% of oocytes, respectively. The rate of cleavage of oocyte over 2-cell stage was 4.1%(10 of 242), These results indicate that the microinjection may be a useful technique to study sperm-oocyte interaction.

• Applied Microscopy
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• v.1 no.1
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• pp.35-42
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• 1969

가토간장에서 채취(採取)한 간(肝)디스토마 성충(成蟲)의 정소(精巢)와 수정낭(受精囊)을 1.25% glutaraldehyde와 1% 사산화(四酸化) 오스뮴산(酸)으로 냉실(冷室)에서 이중고정(二重固定)하였다. 고정(固定)된 시료(試料)는 양식(樣式)에 따라 탈수(脫水)한 후(後) Epon-812로 포매(包埋)하여 MT-2형(型) Porter Blum microtome으로서 초박절편(超薄切片)을 만들어 수사화연(水酸化鉛)과 초산(醋酸)우라닐로서 이중염색(二重染色)한후 Hitachi HS-7형(型)과 HU-11E형(型)의 전자현미경(電子顯微鏡)으로 관찰(觀察)하였다. 관찰결과 간(肝)디스토마의 정세포(精細胞)는 타원형으로 난형(卵形)의 인(仁)을 포함(包含)한 큰 핵(核)을 갖고 있으며 핵(核)을 둘러싼 비교적소량(比較的小量)의 세포질(細胞質)에는 낭형(囊形)의 조면소포체(粗面小胞體)가 희소(稀少)하게 있으며 유리(遊離)리보좀 및 즐(櫛)을 가진 미토콘드리아, 중심체(中心體), 층판상(層板狀)의 골지체가 있다. 정자(精子)는 긴 원주형(圓柱形)의 핵(核)과 선단(先端)에 첨체(尖體)를 포함(包含)한 두부(頭部)와 중종편(中終片), 미부(尾部)의 말단부(末端部)로 갈수록 점차 가늘어져서 결국(結局) 편모(鞭毛)로 끝난다. 두부(頭部)의 선단부(先端部) 가까이 핵환(核環)이 있으므로 이를 기시점(起始點)으로 직경(直徑) $250{\AA}$ 정도의 $8{\sim}10$개(個)의 미세소관(微細小管)이 축사(軸絲)의 배면(背面) 원형질막(原形質膜) 직내면(直內面)의 외형질(外形質)에 평행(平行)하게 중종편(中終片)까지 신장(伸長)되어 있다. 미토콘드리아는 융합(融合)되어 두부(頭部)의 후반부(後半部) 핵(核)의 복측(腹側)에 평행(平行)하게 축사(軸絲)를 감싸는 원추형(圓錐形)으로 종단면(縱斷面)에서 반점상(班點狀)의 횡문(橫紋)이 관찰(觀察)되었다. 중심체(中心體)에서 기원(起原)된 축사(軸絲)는 중심부(中心部)에 중심섬유(中心纖維)와 중심초 및 이중(二重)의 미세소관(微細小管)이 9개(個) 둘러싸고 있는 구조(構造)를 하고 있다.

• Applied Microscopy
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• v.28 no.1
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• pp.63-71
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• 1998

The spermatozoon of Squalidus gracilis majimae is approximately $36.6{\mu}m$ in length and is characterized by a spherical nucleils with the clear chromatin, a short midpiece containing the mitochondria, and a flagellum positioned tangentially to the nucleus. An acrosome is absent as in all teleost fishes. The nucleus is about $1.9{\mu}m$ in diameter and in its periphery contains the electron-lucent chromatin distinguished from the electron-dense chromatin occupying most of the nucleus. The shallow nuclear fossa contains the proximal centriole, instead of two centrioles in deep nuclear fossa in siluroids. The proximal and distal centrioles are oriented approximately $140^{\circ}$ to each other. The mitochondria of 10 or more in number are arranged in three layers and do not surround the axoneme. The asymmetrical distribution of the mitochondria and the eccentrical position of the nucleus with regard to the tail are the general pattern of the cyprinid spermatozoa. S. gracilis majimae spermatozoa have the most mitochondria and the deepest cytoplasmic canal among cyprinid species. The flagellum lacks the lateral fins.

• Applied Microscopy
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• v.41 no.1
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• pp.31-35
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• 2011

The fine structures of the sperm morphology in the Myotis daubentonii ussuriensis were observed by transmission electron microscope. The results showed that the sperm head revealed bullet shaped, the width was showed a slender more than toward the posterior region to anterior region of nucleus. The sperm head was about $4.5{\mu}m$ in length, being about $2.0{\mu}m$ in width. The nuclear length was $4.3{\mu}m$, occupied most of the sperm head. The nucleus and acrosome were separated by the apical body. The neck region was composed the basal plate, capitulum and segmented columns. The segmented columns were about 12 to 14 in number and connected with the outer dense fibers of the middle piece. The mitochondria sheath were arranged like the thread of a screw, and the total number of mitochondrial gyres were 57. The satellite fibers were observed irregularly among the outer dense fibers in the middle piece. Except the middle piece they are not observed in the principal and end pieces of the tail. In general, the tail show axoneme composed of a 9+2 microtubular pattern, and microtubules of the end piece were arranged irregularly.