• Journal of Radiation Protection and Research
• /
• v.10 no.1
• /
• pp.29-40
• /
• 1985

The objectives of present study is to investigate genetic damage of radiation in mammalian male germ cell and. to establish available screening method for determining genetic hazard by radiation. Several methods were employed to measure the genetic damage of radiation as follows: Sperm head counts, frequency occurrence of sperm with abnormal head shape, fertility, activity of LDH-X, and the induction of unscheduled DNA synthesis (U.D.S.) in male mouse were performed with the passing of time after irradiation by making use of the sequence of event that occurs during spermatogenesis. Sperm head counts and activity of LDH-X in testes were gradually reduced by increased radiation dose and with the passing of the time after irradiation. Frequency occurrence of sperm with abnormal head shape, sterile period, and the induction of unscheduled DNA synthesis were increased by increased radiation dose. It is suggested that since germ cell is a direct reflection of genetic complement, the use of male germ cell is rapid and convenient method for measuring genetic damage by radiation.

• The Korean Journal of Zoology
• /
• v.38 no.4
• /
• pp.514-522
• /
• 1995

In order to examine the interaction of albumin with the sperm during capacitation in mouse, proteins of cauda epididymal sperm were extracted under various conditions and analyzed with SDS-PAGE. Sperm surface labeling patterms were also examined using fluorochroin~conjugated wheat germ agglutinin (WGA) and bovine serum albumin (BSA). Albumin was detached from the sperm surface during the incubation and seemed to be constituted the major protein components of the conditioned media in which sperm incubated for 90 mm. Detachment of albumin from the sperm was not affected by the Ca2+ in the medium. WGA-FITC labeling confirmed that Triton X-100 permeabilired plasma membrane overlaying the apical segment of sperm head and detached plasma membrane associated proteins having negatively charged glycoconjugates. BSA-FITC labeling of epididymal sperm occurred on the apical segment of periacrosoinal region and postacrosomal region of the head. BSA-FITC labeling was not observed in periacrosoinal region of the sperm treated with Ca2+-ionophore ~3187 (10 MM)~ whereas the postacrosome region of acrosome-reacted sperm was still labeled after the AR. These results suggest that albumin bound to the surface of epididymal sperm is detached during the capacitation process, and it might be involved In physiological change of sperm plasma membrane accompanying the capacitation.

• Clinical and Experimental Reproductive Medicine
• /
• v.49 no.1
• /
• pp.40-48
• /
• 2022

Objective: As the associations of sperm DNA fragmentation with morphology have not been examined in detail, this study aimed to investigate the relationship between abnormalities of morphological details and DNA integrity in human sperm. Methods: In this cross-sectional study, men from infertile couples were enrolled at Hue Center for Reproductive Endocrinology and Infertility, Vietnam. Conventional semen parameters, including morphological details, were analyzed following the World Health Organization 2010 criteria. Sperm DNA fragmentation was evaluated using a sperm chromatin dispersion assay. The relationships and correlations between semen parameters, sperm morphology, and the type of halosperm and the DNA fragmentation index (DFI) were analyzed. Results: Among 130 men in infertile couples, statistically significant differences were not found in the sperm halo type between the normal and abnormal sperm morphology groups. The percentage of round-head spermatozoa was higher in the DFI >15% group (16.98%±12.50%) than in the DFI ≤15% group (13.13% ±8.82%), higher values for amorphous heads were found in the DFI >15% group, and lower values for tapered heads were observed in the DFI ≤15% group; however, these differences were not statistically significant. Small-halo sperm and the DFI were positively correlated with round-head sperm (r=0.243, p=0.005 and r=0.197, p=0.025, respectively). Conclusion: The rate of general sperm morphological abnormalities in semen analysis was not related to sperm DNA integrity. However, round sperm heads were closely associated with sperm DNA fragmentation.

• Clinical and Experimental Reproductive Medicine
• /
• v.46 no.2
• /
• pp.60-66
• /
• 2019

Objective: To compare black rice (Oryza sativa L) extract with three different staining methods for human sperm head assessment. Methods: Semen samples were collected from 34 volunteers. Four smears of each ejaculate were prepared for staining using the rapid Papanicolaou (PAP) stain, SpermBlue, DipQuick, and black rice extract. The percentage of defective sperm heads (mean${\pm}$standard deviation) was compared. Results: Black glutinous rice extract, a natural dye, was used instead of hematoxylin to stain the nuclei of the sperm heads. The percentage of defective sperm heads showed a significant difference between black rice extract and DipQuick (p= 0.000). In contrast, black rice extract and rapid PAP showed no statistically significant difference (p= 0.974). A strong correlation (r = 0.761) was found between the findings obtained using rapid PAP and black rice extract. In contrast, a weak correlation (r = 0.248) was obtained between DipQuick and black rice extract for the percentage of defective sperm heads. Conclusion: The results showed good agreement and a strong correlation between the rapid PAP and black rice extract stains. The advantages of black rice extract as a novel substitute for hematoxylin for nuclear staining include ease of preparation, local availability, and favorable nuclear staining properties. Further studies could also focus on comparing staining techniques in clinical samples.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.6
• /
• pp.755-759
• /
• 2004

The present investigation was carried out to assess the effect of Sephadex (G-15) filtration on the post thaw bull semen quality and conception rate. Post thaw unfiltered (control) and Sephadex filtered semen from four healthy bulls (three cross bred and one pure bred Holstein Friesian) were subjected to microscopic examination viz. sperm concentration, individual motility, live sperm count and sperm morphology. Sixty-two healthy, normal cycling crossbred cows were inseminated with post thaw unfiltered (n=32) and filtered semen (n=30). Sephadex filtration of post thaw semen significantly (p<0.05) decreased total sperm concentration and sperm with abnormal head, mid piece and tail. The overall average total sperm concentration, head and tail defects in filtered semen decreased significantly (53.4, 1.2 and 6.4 million) than in the unfiltered semen (80.4, 2.4 and 15.7 million, respectively). However, after filtration significant (p<0.05) increase in overall average motile and live sperm concentration were observed (38.8 and 38.0) as compared to unfiltered semen (29.2 and 32.0 million, respectively). The overall conception rate recorded was 21.9% with post thaw unfiltered semen and 56.7% with filtered semen. It was concluded that Sephadex filtration of post thaw semen improved its quality and conception rate.

• Applied Microscopy
• /
• v.40 no.4
• /
• pp.253-259
• /
• 2010

The ultrastructure of sperms in testes of the pond smelt (Hypomesus nipponensis) was investigated using electron microscopes. The whitish testis was located between swim bladder and intestine. Especially, the left testis was larger than the right testis. The sperm was approximately $26\;{\mu}m$ in length. The sperm had an oval head and the acrosome was not found. The nucleus was about 400 nm in diameter and chromatin was incompletely condensed. The nuclear fossa deeply formed in sperm head and two centrioles were located in the fossa. The mitochondrium was observed only one in midpiece of the sperm and a motile flagellum consisted of an axoneme with a typical 9+2 pattern of microtubule. Also, the tail of the sperm has axonemal fins.

• Journal of Embryo Transfer
• /
• v.16 no.1
• /
• pp.61-68
• /
• 2001

Sperm competent for fertilization can become capacitated, bind to the zona pellucida (ZP) of an egg in a specific manner, and complete acrosomal exocytosis. Failure to carry out these functions results in infertility. Although the interactions between the ZP and the plasma mem-brane overlying the sperm acrosome have been considered important for sperm-egg recognition and signalling, recent results have prompted a reassessment of current paradigms concerning these interactions. In this review, we're going to discuss about the roles of the acrosomal matrix, the particulate component of the acrosomal contents, in fertilization. The general hypothesis is that acrosomal exocytosis leads to the exposure of acrosomal matrix proteins that become de-facto extracellular matrix (ECM) on the surface of the sperm head, and that the dynamic interactions of this newly-exposed sperm ECM with the egg ECM (the ZP) govern sperm-egg recognition and sperm penetration of the ZP Informations from these experiments may provide new ways to address the poor ZP binding of sperm from some human infertility patients and may offer new avenues for contraception through the disruption of purposeful sperm-ZP binding.

• Proceedings of the KSAR Conference
• /
• 2001.03a
• /
• pp.12-12
• /
• 2001

To get insight into the nature of foreign mitochondria and syngamy during mammalian fertilization we compared fertilization processes in porcine oocytes following microinjection of porcine or mouse spermatozoa. Pronuclear movement, sperm mitochondria, and DNA synthesis were imaged with propidium iodide, mitotracker, and BrdU under confocal laser scanning microscope. Intracytoplasmic injection of either porcine or mouse spermatzoon activated porcine oocytes without additional parthengenetic stimulation. Foreign mitochondria in either mouse or porcine sperm midpiece were introduced into porcine oocytes following sperm injection, but rapidly disappeared from the actively developing porcine oocytes. BrdU experiment showed new DNA synthesis in porcine oocytes following injection of mouse spermatozoon or sperm head. At 24 h after injection of mouse isolated sperm head or a spermatozoon, mitoic metaphase was seen in oocyte, but they did not go to normal cell division (Table). These results suggest that pronuclear formation, foreign mitochondria disruption, DNA synthesis and syngamy formation during fertilization are not species specific processes.(Table Omitted).

• Journal of Aquaculture
• /
• v.12 no.1
• /
• pp.57-62
• /
• 1999

Experiments were performed to study the activity and fertility of grey mullet (Mugil cephalus) sperm after the courses of cold storage and cryopreservation. The head of spermatozoon showing spherical shape was sized $1.26{\pm}0.08 \{mu}textrm{m}$ in diameter and its nucleus contained numerous granular chromatins. Flagellum of tail showed typical 9＋2 structure. Preservation of grey mullet sperm was the most effective when it was stored with serum of the same species at $0^{\circ}C$ and sperm activity index was similar in egg-tris, 0.1 M, 0.3 M and 0.5 M glucose. When grey mullet sperm were cryopreserved in MFRS as diluent with 10% dimethyl sulfoxide was effective compared with other diluents. Some of post-thawed spermatozoa showed the enlarged head and ruptured plasma membrane compared with unfrozen spermatozoa.

• Applied Microscopy
• /
• v.41 no.2
• /
• pp.117-122
• /
• 2011

Sperm morphology of Pipistrellus savii velox was examined by transmission electron microscope. The sperm head of P. savii velox was bullet in shape. The sperm head was 3.1 ${\mu}m$ in length, whose posterior 3.0 ${\mu}m$ was occupied by a nucleus with 1.8 ${\mu}m$ in width. The segmented columns were about 14~15 in number. The total number of mitochondrial gyres was 57. Number of 1, 5, and 6 of the outer dense fibers were larger than the others. A fibrous sheath and longitudinal column of the principal piece were evidence, but the fibrous sheath was not seen at the end piece. In the present study, the length of the sperm head of P. savii velox were very shorter than those of other bats.