• Asian-Australasian Journal of Animal Sciences
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• v.31 no.10
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• pp.1591-1597
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• 2018

Objective: Selenium-independent glutathione peroxidase (GPx5) is specifically expressed in the mammalian epididymis and plays an important role in protecting sperm from reactive oxygen species and lipid peroxidation damage. This study investigates GPx5 expression in the epididymis of Small Tail Han sheep. Methods: GPx5 expression was studied in three age groups: lamb (2 to 3 months), young (8 to 10 months), and adult (18 to 24 months). The epididymis of each age group divided into caput, corpus and cauda, respectively. Analysis the expression quantity of GPx5 in epididymis and testis by real-time fluorescent quantitative polymerase chain reaction and Western blot. Finally, GPx5 protein locating in the epididymis by immunohistochemical. Results: The results demonstrate that in the lamb group, the GPx5 mRNA, but not protein, can be detected. GPx5 mRNA and expressed protein were detected in both the young and adult groups. Moreover, both the mRNA and protein levels of GPx5 were significantly higher in the young group than in other two groups. When the different segments of epididymis were investigated, GPx5 mRNA was expressed in each segment of epididymis regardless of age. Additionally, the mRNA level in the caput was significantly higher than that in corpus and cauda within same age group. The GPx5 protein was in the epithelial cells' cytoplasm. However, GPx5 mRNA and protein were not detected in the testis. Conclusion: These results suggest that GPx5 is mainly expressed in the epididymis of Small Tail Han sheep, and that the expression level of GPx5 is associated with age. Additionally, GPx5 was primarily expressed in the epithelial cells of the caput. Taken together, these studies indicate that GPx5 is expressed in the epididymis in all age grades.

• Korean Journal of Ecology and Environment
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• v.35 no.3 s.99
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• pp.232-236
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• 2002

In an effort to uncover the reproduction of Korean brook perch Coreoperca herzi testis anatomy and sperm morphology were studied. Fish samples were collected in the Sooypcheon river from May to October 2001. White-colored testes have wedgeshaped external morphology, and developed symmetrically in the dorsal cavity of the trunk. Isogenetic germ cells developed in the cyst located in seminiferous lobule. Each lobule showed significant asynchrony in the spermatogenic stage of the cyst. Sperm was 43 ${\mu}$m in length. The round head was 2.2 ${\mu}$m long. The middle piece developed beneath the head was 0.5 ${\mu}$m long. Tail was 40 ${\mu}$m in length. Coomassie brilliant blue (CBB) gave rise the intense staining in the apex of sperm head and middle piece, suggesting the possible development of acrosome.

• Korean Journal of Animal Reproduction
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• v.17 no.4
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• pp.339-346
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• 1994

An understanding of the structure and function of mammalian spermatozoa requires the iso-lation of these components. In this study, frozen-thawed bovine spermatozoa were treated by physical treatments (vortexing, 26 gauge needle, strained 26 gauge needles and freezing-thawing) or chemical treatments (trypsin, dithiothreitol, sodium dodecylsulfate and $\beta$-mercaptoethanoJ) to yield free heads and tails. The most effective treatment was repeated pumping of sperm suspension through a strained 26 gauge needle conneted to a syringe. Spermatozoa by this treatment were mainly broken at the junction of the head and the tail, resulting in 90-100% yields. Also, sperm head surface did not modify during strained 26 gauge needle treatment when either spermatozoa or sperm heads were incubated in 250${\mu}\textrm{g}$/ml of FITC-UEA 1 for 1 h at room temperature to detect the modification of sperm surface components. Other physical treatments were less efficient for the breakdown of spermatozoa. The effects of chemical treatments on bovine spermatozoa are not noticeable. Dissected sperm heads and tails should be fractional leading to nearly pure components by sucrose gradient centrifugation at 1,000 rpm for 15 min. The result suggest that the established method may be useful for the biochemical study of spermatozoal components, and the understanding of oocyte activation mechanism either by spermatozoal components during fertilization or microinjection of isolated components.

• Applied Microscopy
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• v.39 no.3
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• pp.227-236
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• 2009

The ultrastructure of spermatogenesis and sperm in Coreoleuciscus splendidus, belonging to Gobioninae, Cyprinidae was investigated by light and electron microscopes. The testis was located between intestine and air bladder. The size of testis was major axis 1.8 cm, minor axis 3 mm. The testis of C. splendidus contained numerous testicular cysts, and spermatogenesis was non-synchronized in these testicular cysts. In May, the upper area of testis contained with other germ cells and sperm but the lower area of testis contained with matured sperm only. In case of spermatogonia, the nucleus was comparatively large spherical, 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 in a middle piece. The head of matured sperm was a spherical shape and had not acrosome. The microtubules of flagella were arranged 9+2 structure. Also, the tail of sperm had not lateral fins and 7 outer coarse fibers.

• Applied Microscopy
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• v.31 no.4
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• pp.333-345
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• 2001

The mature sperms of three species of cephalopods (Octopus minar, Octopus ocellatus, Todarodes pacificus) were observed by electron microscopy. The results obtained are as follows: The sperm lengths of Octopus minor and Octopus ocellatus of octopods are long and they are about $390{\mu}m$ and $125\sim130{\mu}m$, respectively, but the sperm length of Todarodes pacificus is short and about $35{\mu}m$. The sperm of Octopus minor has a helical acrosome and a head bent a little like a banana while Octopus ocellatus of octopod has a twisted acrosome and a long rod-shaped head. A number of horizontal stripes are observed as a periodic structure in their subacrosome cavities and dense plugs are formed in the cavities of their heads. On the other hand, the acrosome of Todarodes pacificus is circular cap-shaped, and its head is long and oval. It is notable that two small cavities were observed in its basal acrosome. Juxtanuclear acrosomal materials of high electron density filled the subacrosomal cavity. In the middle piece of mature sperms of Octopus minor and Octopus ocellatus, the mitochondria form the mitochondrial sleeve, but the numbers of mitochondria differ between the species so that they are $11\sim12$ and $8\sim9$, respectively. Meanwhile, in the middle piece of mature sperms of Todarodes pacificus, the mitochondria are separated from the axoneme, forming a mitochondrial spur in which $10\sim13$ mitochondria and some electron dense materials concentrate. The axoneme of Octopus minor, Octopus ocellatus and Todarodes pacificus are of 9+2 type in common, surrounded by 9 coarse fibres. A number of glycogen were observed only in the axoneme of Todarodes pacificus. The coarse fibres were found as far as the main piece of sperm tail in Octopks minor and Todarodes pacificus, while to the end piece of sperm tail in Octopus ocellatus.

• Development and Reproduction
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• v.1 no.2
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• pp.133-139
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• 1997

To study the function and structure of Golgi apparatus in the spermiogenesis of long-fingered bat (Miniopterus schreibersi fuliginosus), the testis obtained from adult bat was treated with the prolonged osmification or fixed with ferrocyanide reduced osmium. golgi apparatus was oval shape in early Golgi phase, and was composed of cortex and medullar enclosing acrosome in mid Golgi phase. The vesicles of crescent shape Golgi apparatus were closed or fused with small or large vesicles at the periphery of acrosome. Golgi apparatus moved behing the acrosome face in cap phase, but the Golgi apparatus was still active. According to this, Golgi apparatus appears to be involved in the formation of acrosome and sperm tail. Transfer of materials from Golgi to acrosme seems to be carried out not only by fusion of large vesicles with acrosomal vesicles but also by detachment of coated vesicle from various cisternae of Golgi fusing with acrosomal vesicle.

• Applied Microscopy
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• v.40 no.1
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• pp.1-8
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• 2010

The ultrastructure of spermatogenesis and sperm in Chinese minnow, Rhynchocypris oxycephalus belonging to Leuciscinae was investigated by light and electron microscopes. The whitish testis was located between intestine and air bladder. The size of testis was major axis 2.3 cm, minor axis 6 mm. The testis contained numerous testicular cysts, and spermatogenesis was non-synchronized in these testicular cysts. In the case of spermatogonium, the nucleus was comparatively large ellipsoidal, and mitochondria showed a marked development. The size of primary spermatocyte was smaller than that of spermatogonia, and secondary spermatocyte was smaller than 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 in a middle piece. The sperm was formed by loss of cytoplasm. The head of mature sperm was a spherical shape and have not acrosome. The microtubules of flagella were arranged 9+2 structure. Also, the tail of sperm have not lateral fins.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.155-162
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• 2005

The present study evaluated whether an exogenous antioxidants, taurine and $\alpha$-tocopherol, could, when added to the freezing extender, improve the post-thaw sperm characteristics, function, the level of reactive oxygen species (ROS) generation, and the level of lipid peroxidation (LPO) in frozen-thawed porcine semen. CASA (computer-aided sperm analysis), HOST (hypoos-motic swelling test), chemiluminescence using luminol and lucigenin and the detection of malondialdehyde for LPO was performed in frozen-thawed porcine sper-matozoa. The results obtained in these studies are as follows. While no beneficial effects of taurine and $\alpha$-tocopherol supplementation were visible in motility, viability, acrosome reaction, tail swelling patterns, and the generation of $O^{2-}$ of frozen-thawed porcine sper-matozoa, $H_{2}O_{2}$ was decreased by all treatments except taurine 50mM treatment. In conclusion the taurine and $\alpha$-tocopherol treatments during freezing reduced generation of reactive oxygen species and production of malondialdehyde in frozen-thawed porcine semen, and the ROS savangers may minimize various damages of spermatozoa during freezing.

• Applied Microscopy
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• v.31 no.3
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• pp.223-233
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• 2001

The spermiogenesis of a Korean octopus, Octopus minor, inhabiting western of Korea Sea was observed by electron microscopy . The obtained results are as follows: The spermiogenesis of Octopus miner proceeds through four stages; early- , mid- , and late-spermatid, and mature sperm. An early spermatid is a spherical cell looking light due to the low electron density. The acrosome formed from Golgi complex of the upper nucleus looks dark due to the high electron density. The extra-nuclear rod (enr) stemming from proximal centriole is transformed from round shape into oval shape, elongating to the upper nucleus. In our observation, the axoneme was being formed from distal centriole, and the manchette composed of a number of microtubules is also found around nuclear membrane. In a mid-spermatid, chromatins in the nucleus contract shaping fine threads, and the manchette is also observed around nuclear membrane. Especially, the spherical acrosome is transformed into long oval one which is tinged with a number of horizontal stripes and has the middle electron density. In a late-spermatid, chromatins in the nucleus contract thick and short. Furthermore, the mitochondrial sleeve, in which the axoneme is surrounded with mitochondria, is observed at middle piece. The axoneme has a typical structure of 9+2 and around it, 9 coarse fibers are observed. Also in the acrosome cavity of mature sperm, horizontal striation is found. However, regularly spaced processes are peculiarly observed in there. A sperm is about 390 fm long, whose head is bent a little like a banana while the acrosome region is helical. In the middle piece of sperm, $11\sim12$ mitochondria are surrounding coarse fibers that reach the main piece of tail, while nothing but 9+2 structured axoneme is found in the end piece.

• Journal of Embryo Transfer
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• v.28 no.1
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• pp.31-39
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• 2013

Alpha-tocopherol as an antioxidant acts in preservation of chilled semen by preserving cell membrane damage from lipid peroxidation. Optimum concentrations of ${\alpha}$-tocopherol in egg yolk-citrate (EYC) extender need to be studied in crossbred bull's semen. Different concentrations of ${\alpha}$-tocopherol viz. 0, 1, 2, 4 and 6mg per ml of extender were used. Semen was collected once a week from four bulls used to regular collection, aged 4 to 7 years, weighing 320 to 450 kg, and with body condition score 4 to 4.5 and scrotal circumference 23 to 32 cm. Semen was evaluated routinely and sperm morphology was viewed under light microscope at ${\times}1,000$ magnification after fixing with buffered formal saline. Over 90% had normal head, acrosome, mid-piece and tail. Semen was diluted with egg-yolk-citrate extender to produce $15{\times}10^6$ spermatozoa/ml and 0, 1, 2, 4 and 6 mg/ml ${\alpha}$-tocopherol were added. The semens amples were kept at $8^{\circ}C$. Sperm motility and viability were examined daily up to 5 days under light microscopy at ${\times}200$ magnification. Sperm viability was acceptable (${\geq}40%$) up to the $4^{th}$ day with all concentrations of ${\alpha}$-tocopherol and up to the $5^{th}$ day with 2 mg/ml ${\alpha}$-tocopherol. Sperm motility was acceptable (${\geq}40%$) up to the $3^{rd}$ day irrespective of ${\alpha}$-tocopherol concentration, and up to the $4^{th}$ day with 2 mg/ml ${\alpha}$-tocopherol. It is suggested that the lifespan of chilled semen may be extended up to 4 days by adding 2mg/ml ${\alpha}$-tocopherol.