• Korean Journal of Veterinary Research
• /
• v.24 no.2
• /
• pp.245-266
• /
• 1984

As a fundamental study to increase the reproductive efficiency in cattle, highly motile sperm were separated and collected from raw semen, extended semen and frozen semen by different methods using various concentrations of bovine serum albumin or Tyrode's solution. Various characteristics and light microscopic and electron-microscopic morphology of sperm separated by different methods were compared. The results obtained were as follows; 1. The sperm separated from raw semen using bovine serum albumin showed significantly high value in motility, motile sperm count, percent of normal sperm and progressive motility, as compared with control sperm and revealed the highest sperm recovery rate when separated with 6% bovine serum albumin. 2. The sperm motility, percent of normal sperm and progressive motility of the highly motile sperm frozen after being separated from raw semen with bovine serum albumin, showed significantly high value than those of a control sperm and especially found the highest value when separated with 20% bovine serum albumin. 3. Light-microscopic percent of abnormality was significantly low in the prefrozen and postfrozen highly motile sperm separated with bovine serum albumin, as compared with control sperm. 4. Electron-microscopic finding of the highly motile sperm separated with bovine serum albumin showed low percent of deformity in the dilatation and vesiculation of cell membrane, in dilatation and density loss of acrosome than in those of control sperm. 5. It was impossible to separate the highly motile sperm from frozen semen with bovine serum albumin, but it was possible with Tyrode's solution. 6. Recovery rate of highly motile sperm from raw semen extended semen and frozen semen was the highest when the sperm pellet stood in Tyrde's solution for 80 minutes. 7. The highly motile sperm separated from raw semen, extended semen and frozen semen with Tyrode's solution showed significantly high value in motility, progressive motility and percent of normal sperm, as compared with control sperm. 8. Highly motile sperm, when separated from raw semen, extended semen and frozen semen with Tyrode's solution, showed significantly low percent of microscopic abnormality as compared with control sperm.

• Journal of Embryo Transfer
• /
• v.27 no.1
• /
• pp.45-50
• /
• 2012

The objective of this study is to estimate the effect of adding TES to LEY and FGE freezing extender for the sperm viability, acrosomal morphology and DNA fragmentation from miniature pig sperm, we evaluated sperm characteristics in TFGE, TLE and LEY with various thawing condition ($37^{\circ}C$ for 20 sec, 45 sec and $75^{\circ}C$ for 5 sec, respectively), and in different concentration of glycerol at 1%, 1.5%, 3%. The sperm viability and normal acrosome intact(NAI) in TFGE (Viability : $60.3{\pm}2.4$, NAI : $58.6{\pm}2.2%$), TLE ($61.3{\pm}2.4$, $62.2{\pm}2.2%$) extender significantly(p<0.05) increased than that in LEY ($50.2{\pm}2.4$, $54.5{\pm}2.2%$) extender thawed at $75^{\circ}C$ for 5 sec. According to the results from glycerol concentration, the viability and NAI of miniature pig sperm in 1.5% glycerol TLE ($66.1{\pm}3.2$, $66.2{\pm}1.0%$) was highest among the experimental groups. In accordance with this, DNA fragmentation rates was the lowest in TLE ($43.3{\pm}0.5%$) while that in LEY ($63.5{\pm}2.3%$) is the highest. Therefore, these results suggest that TLE extender method for freezing- thawing of miniature pig sperm increased the viability after thawing.

• Journal of Life Science
• /
• v.11 no.3
• /
• pp.218-229
• /
• 2001

The cycle of the seminiferous epithelium and spermiogenesis in three species if the genus Crocidura, the lesser white-toothed shrew, C. suaveolens, the Japanese white-toothed shrew, C. dsinezumi and the big(=Ussuri) white-toothed shrew C. lasiura, in the breeding season were studied with light and electron microscopes. The three species examined are distinguished from each other in the morphology of the seminiferous epithelium and the spermiogenesis, suggesting that these morphological characteristics are useful for the identification of the species. C. dsinezumi and C. lasiura, however, share many characteristics which are not common in C. suaveolens, as follows: In both species, 1) the cycle of the seminiferous epithelium is composed of 10 stages against 11 stages in C. suaveloens; 2) the earliest intermediated type spermatogonia is observed at stage I against stage III in C. suaveolens; 3) the spermatids of step 5 is observed during the stages V-VI against stages V-VII in C. suaveolens; 4) the acrosomal extension occurs during the stages VIII-X against tages IX-XI in C. suaveolens; 5) the condensation of the nucleus occurs simultaneously whereas it begins from the middle and along the nuclear membrane in C. suaveolens; 6) the capitular length in acrosome phase is shorter(about 2/3 of the diameter of the proximal centriole) than in C. suaveolens(longer than the diameter of proximal centriole; 7) length of the post nuclear cap is shorter(less than a half of the nucleus) than in C. suaveolens(about a half on the nucleus). Hudging from the similarities in the spermatogenesis in C. dsinezumi and C. lasiura, the relationship between them seems to be close compared to those with C. suaveolens.

• Development and Reproduction
• /
• v.14 no.4
• /
• pp.269-279
• /
• 2010

Some characteristics of germ cell differntiations and the function of accessory cells during spermatogenesis, and mature sperm ultrastructure in male Protothaca (N.) jedoensis were investigated by transmission electron microscope observations. The morphology of the spermatozoa of this species has a primitive type and is similar to those of other species in the subclass Heterodonta. Accessory cells, which are connected to adjacent germ cells, are involved in the supplying of the nutrients for germ cell development. The morphologies of the sperm nucleus and the acrosome of this species are the cylindrical type and cap shape, respectively. Spermatozoa are approximately $46{\sim}50{\mu}m$ in length including a long sperm nucleus (about $2.44{\mu}m$ in length), an acrosome (about $0.45{\mu}m$ in length), and tail flagellum (about $42{\sim}46{\mu}m$). The axoneme of the sperm tail shows a 9+2 structure. As some characteristics of the acrosomal vesicle structures, the basal and lateral parts of basal rings show electron opaque part (region), while the anterior apex part of the acrosomal vesicle shows electron lucent part (region). These characteristics of the acrosomal vesicle were found in the family Veneridae and other several families in the subclass Heterodonta. These common characteristics of the acrosomal vesicle in the subclass Heterodonta can be used for phylogenetic and systematic analysis as a taxonomic key or a significant tool. The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appear in most species in the family Veneridae and other families in the subclass Heterodonta. However, exceptionally, only three species in Veneridae of the subclass Heterodonta contain 5 mitochondria. The number of mitochondria in the sperm midpiece can be used for the taxonomic analysis of the family or superfamily levels as a systematic key or an important tool.

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

• Development and Reproduction
• /
• v.13 no.1
• /
• pp.25-33
• /
• 2009

The cycle of the seminiferous epithelium and development of spermatids of Apodemus speciosus peninsulae were observed using a light microscope. On the basis of developing spermatocyte and spermatid, the cycle of the seminiferous epithelium was divided into 9 stages. Type Ad spermatogonia were appeared in all stages ($I{\sim}IX$). The Ap, In, and B types of spermatogonia were appeared from stage I, II and III, and IV, respectively. In prophase of first meiosis, the leptotene spermatocytes appeared from stage V and VI, zygotene spermatocytes from stages I, II, VII, VIII, and IX, pachytene spermatocytes from stage III to VII, diplotene spermatocytes in the stage VIII, and secondary spermatocytes in stage IX. On the basis of morphology of spermatid head, developing of nuclear and acrosome and the morphological change of cytoplasm, the developing of spermatids was divided into 12 steps. Considering all the results, A. s. peninsulae displayed very similar result with A. agrarius coreae that is allied species when compare correct characters developing of spermatids with spermatogonia and appearance time of the spermatocyte. Appearance time of the same cell and number of spermatogonial generation was thought that characters of the species, and information may be useful in identifying the species.