• The Korean Journal of Malacology
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• v.27 no.2
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• pp.149-157
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• 2011

Some characteristics of the formations of acrosomal vesicles during the late stage of spermatids during spermiogenesis and taxonomical charateristics of sperm morphology in male two species (Saxidomus purpurata and Meretrix petechialis) in the family Veneridae were investigated by electron microscope observations. In two species, the morphologies of the spermatozoa have the primitive type and are similar to those of other bivalves in that it contains a short midpiece with five mitochondria surrounding the centrioles. The morphologies of the sperm nuclear types of S. purpurata and M. petechialis in Veneridae have the curved cylindrical and cylinderical type, respectively. And the acrosome shapes of two species are the same cap-shape type. In particular, the axial filament is not found in the lumen of the acrosome of two species, however, subacrosomal material are observed in the subacrosomal spaces between the anterior nuclear fossa and the acrosomal vesicle of two species. The spermatozoon of S. purpurata is approximately 46-$52{\mu}m$ in length, including a curved sperm nucleus (about $3.75{\mu}m$ in length), a long acrosome (about $0.40{\mu}m$ in length),and a tail flagellum (about 45-$47{\mu}m$ long). And the spermatozoon of M. petechialis is approximately 47-$50{\mu}m$ in length including a slightly curved sperm nucleus (about $1.50{\mu}m$ in length), an acrosome (about $0.56{\mu}m$ in length) and tail flagellum (44-$48{\mu}m$ in length). In two species, the axoneme of the sperm tail flagellum of each species consists of nine pairs of microtubules at the periphery and a pair of cental doublets at the center. Therefore, the axoneme of the sperm tail flagellum shows a 9 + 2 structure. In particular, taxonomically important some charateristics of sperm morphologies of two species in the family Veneridae are acrosomal morphology of the sperm, The axial filament is not found in the acrosome as seen in a few species of the family Veneridae in the subclass Heterodonta. The acrosomal vesicle is composed of right, left basal rings and the apex part of the acrosomal vesicle. In particular, right and left basal rings show electron opaque part (region), while the apex part of the acrosomal vesicle shows electron lucent part (region). These charateristics belong to the subclass Heterodonta, unlikely a characteristic of the subclass Pteriomorphia showing all part of the acrosome being composed of electron opaque part (region). Therefore, it is easy to distinguish the families or the subclasses by the acrosomal structures. The number of mitochondria in the midpiece of the sperm of S. purpurata and M. petechialis in Veneridae are five. However, the number of mitochondria in the midpiece of the sperm in most species of Veneridae in the subclass Heterodonta are four. Therefore, the number of mitochondria of the sperm midpiece of two species are exceptionally 5, and it is only exceptional case in the species in Veneridae in the subclass Heterodonta. Except these cases, the number of mitochondria in the sperm midpiece in all families in the subclass Heterodontaare are 4, and now widely used in taxonomic analyses.

• Clinical and Experimental Reproductive Medicine
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• v.13 no.2
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• pp.175-180
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• 1986

It has been reported that oral kallikrein therapy exerts a favourable effect on sperm motility in asthenozoospermic patients. In order to evaluate the efficacy of kallikrein on asthenozoospermia, a total of 20 subfertile male patients with varicocele, whose sperm counts were less than $40{\times}10^6/ml$ and sperm motility was less than 30%, was subjected to this clinical study (Table 1). They were divided into 2 study groups: 1) Varicocelectomy group consisted of 10 patients with varicocele (grade II-III) who underwent varicocelectomy. 2) Kallikrein group was composed of 10 patients with varicocele (grade I) who were given kallikrein orally 600 KU (kallikrein unit) daily divided 3 times after meal for 3 to 9 months. Semen analyses were repeated twice before the study, once a month during the study and twice after the study. Effective results designate that sperm parameters improved more than 30% from the basical levels after varicocelectomy or kallikrein exposure. Sperm counts increased from $32.5{\times}10^5/ml$ to $45.5{\times}10^6/ml$ after varicocelectomy in 3 patients and sperm motility increased from 25% to 38.5% after varicocelectomy in 3 patients. Pregnancy occurred in 2 patients of 3 responders and 1 patient of 7 non-responders 3 to 6 months after varicocelectomy in Varicocelectomy group. Sperm motility increased from 28% to 40.2% after kallikrein treatment in 3 patients. Pregnancy occurred in 2 patients of the 3 responders in Kallikrein group (Tables 2-3). There were no significant changes in volume and morphology in Varicocelectomy group before after varicocelectomy and no significant changes in volume, counts, and morphology before and after kallikrein exposure. No remarkable side effects were noted with kallikrein treatment.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.3
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• pp.185-191
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• 2023

Objective: Although intracytoplasmic sperm injection (ICSI) is a way to deal with in vitro fertilization failure, 3% of couples still experience repeated fertilization failure after attempted ICSI, despite having sperm within normal parameters. These patients are a challenging group whose sperm cannot fertilize the egg during ICSI. Unfortunately, no test can predict the risk of fertilization failure. Phospholipase C zeta (PLCζ) and transition nuclear proteins (TNPs) are essential factors for chromatin packaging during sperm maturation. This study aimed to assess PLCζ1 and TNP1 expression in the sperm of patients with fertilization failure and the correlations among the DNA fragmentation index, PLCζ1 and TNP1 gene and protein expression, and the risk of fertilization failure. Methods: In this study, 12 infertile couples with low fertilization rates (<25%) and complete failure of fertilization in their prior ICSI cycles despite normal sperm parameters were chosen as the case group. Fifteen individuals who underwent ICSI for the first time served as the control group. After sperm analysis and DNA fragmentation assays, quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and Western blot analyses were performed to compare the gene and protein expression of PLCζ and TNP1 in both groups. Results: DNA fragmentation was significantly higher in the fertilization failure group. The qRT-PCR and Western blot results demonstrated significantly lower PLCζ and TNP1 gene and protein expression in these patients than in controls. Conclusion: The present study showed that fertilization failure in normozoospermic men was probably due to deficient DNA packaging and expression of TNP1.

• Development and Reproduction
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• v.16 no.1
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• pp.19-29
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• 2012

The ultrastructural characteristics of germ cell differentiations during spermatogenesis and mature sperm morphology in male $Atrina$ ($Servatrina$) $pectinata$ were evaluated via transmission electron microscopic observation. The accessory cells, which contained a large quantity of glycogen particles and lipid droplets in the cytoplasm, are assumed to be involved in nutrient supply for germ cell development. Morphologically, the sperm nucleus and acrosome of this species are ovoid and conical in shape, respectively. The acrosomal vesicle, which is formed by two kinds of electron-dense or lucent materials, appears from the base to the tip: a thick and slender elliptical line, which is composed of electron-dense opaque material, appears along the outer part (region) of the acrosomal vesicle from the base to the tip, whereas the inner part (region) of the acrosomal vesicle is composed of electron-lucent material in the acrosomal vesicle. Two special characteristics, which are found in the acrosomal vesicle of A. ($S$) $pectinata$ in Pinnidae (subclass Pteriomorphia), can be employed for phylogenetic and taxonomic analyses as a taxonomic key or a significant tool. The spermatozoa were approximately $45-50{\mu}m$ in length, including a sperm nucleus (about $1.43{\mu}m$ in length), an acrosome (about $0.51{\mu}m$ in length), and a tail flagellum (about $46-47{\mu}m$). The axoneme of the sperm tail evidences a 9+2 structure.

• Clinical and Experimental Reproductive Medicine
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• v.51 no.3
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• pp.205-212
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• 2024

Objective: Cyclophosphamide (CP) is an alkylating agent commonly used in cancer treatment. It is known to have detrimental effects on the reproductive system, including the potential to cause infertility. Recently, herbal remedies have gained traction as a complementary approach to addressing these side effects. In this study, our goal was to investigate whether the aqueous-alcoholic extract of Withania somnifera (WS) could mitigate the adverse impacts of CP on testicular tissue. Methods: Animals were randomly assigned to one of the following groups: control, WS (500 mg/kg), CP (100 mg/kg), CP+WS pre-treatment, and CP+WS post-treatment. WS was administered orally through gavage for 1 month. We assessed sperm parameters, testicular histopathology, and the expression of the Bax and Bcl2 genes in the experimental groups. Results: Sperm parameters (including count, viability, and motility), the number of spermatogonia, the seminiferous tubule diameter, and Bcl2 gene expression, significantly decreased after CP injection (p<0.05). Conversely, the number of immotile sperm and Bax gene expression significantly increased (p<0.05). Treatment with WS, especially when administered as a pre-treatment, ameliorated the sperm parameters, histological alterations, and the expression of apoptosis-related genes (p<0.05). Conclusion: The data suggest that WS may mitigate the detrimental effects of CP on testicular tissue by reducing apoptosis. Consequently, WS has the potential to be used as an adjunctive therapy to reduce the complications associated with CP treatment.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.43-43
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• 2001

This study evaluated the effect of fertilization-promoting peptide (FPP) on fertilizing ability and glycosidase activity in vitro of spermatozoa frozen-thawed in pig, Using chlortetracycline fluorescence analysis, the various glycosidase analyses and the oocyte penetration test, we have obtained evidence that FPP can promote the fertilizing ability and glycosidase activity of frozen-thawed spermatozoa in vitro. When frozen-thawed spermatozoa was washed with different concentrations of FPP, there were significantly (P<0.05) more acrosome-reacted in medium with 100 nM than 0, 50, 200 and 400 nM. The penetration rates were also highest in medium containing with 100 nM FPP (P<0.05). On the other hand, the $\beta$-N-acetylglucosaminidase activity was at least twofold higher than other glycosidase. In same glycosidase, however, there were no difference in medium with different concentrations of FPP In another experiment, spermatozoa preincubated in medium with or without FPP for 0, 1, 2, 3 and 4 h were inseminated with oocytes matured in vitro. The percentages of spermatozoa that reached acrosome reaction were affected by preincubation and were higher in medium with that than without FPP. When oocytes were inseminated with spermatozoa preincubated in medium with and without FPP during the different periods, however, penetration rates were decreased with preincubation periods of spermatozoa. On the other hand, when the sperm-oocyte were cultured for 4, 8, 12, 16, 20 and 24 h, the penetration rates were higher in spermatozoa preincubated with that than without FPP and had a tendency to increase as time of culture periods. However, The activities of $\alpha$-fucosidase, $\alpha$ -mannosidase, $\beta$-galactosidase and N-acetyl- $\beta$-D-glucosaminidase were higher in medium with that than without FPP regardless of periods of sperm preincubation and sperm-oocyte culture. These results suggest that FPP may play a positive role in promoting of sperm function and glycosidase activity in vitro in pig.

• Animal Bioscience
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• v.36 no.12
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• pp.1796-1805
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• 2023

Objective: This study aims to identify heat shock protein70-2 (HSP70-2) and protamine-1 (PRM1) mRNA and protein in Madura bull sperm and demonstrate their relation as bull fertility biomarkers. Methods: The Madura bull fertility rates were grouped based on the percentage of first service conception rate (%FSCR) as high fertility (HF) (79.04%; n = 4), and low fertility (LF) (65.84%; n = 4). mRNA of HSP70-2 and PRM1 with peptidylprolyl isomerase A (PPIA) as a housekeeping gene were determined by quantitative real-time polymerase chain reaction, while enzyme-linked immunoassay was used to measure protein abundance. In the post-thawed semen samples, sperm motility, viability, acrosome integrity, and sperm DNA fragmentation index were analyzed. Data analysis was performed on the measured parameters of semen quality, relative mRNA expression, and protein abundance of HSP70-2 and PRM1, among the bulls with various fertility levels (HF and LF) in a one-way analysis of variance analysis. The Pearson correlation was used to analyze the relationship between semen quality, mRNA, proteins, and fertility rate. Results: Relative mRNA expression and protein abundance of HSP70-2 and PRM1 were detected and were found to be highly expressed in bulls with HF (p<0.05) and were associated with several parameters of semen quality. Conclusion: HSP70-2 and PRM1 mRNA and protein molecules have great potential to serve as molecular markers for determining bull fertility.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.2
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• pp.135-139
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• 2003

Objective: We investigated whether serum testosterone to estradiol ratio was decreased in infertile men and whether this condition can be corrected with oral aromatase inhibitor. Method: The serum testosterone to estradiol ratio of 26 men with testicular failure were compared with those of normal semen analysis parameter, 89 control reference group. All of 26 testicular failure group were diagnosed with the previous testicular biopsy. Then 46 men with oligospermia and/or asthenospermia were selected and treated with 1 mg of the aromatase inhibitor anastrozole ($Arimidex^{(R)}$) orally once daily for 3 months. Testosterone to estradiol ratio and semen analyses were evaluated during anastrozole therapy. Results: The testosterone level of testicular failure group was significantly lower and the testosterone to estradiol ratio was more decreased than normal semen parameter group. Forty six on-anastrozole group had significantly lower testosterone (4.6 versus 5.7 ng/ml, p<0.01) and higher estradiol (15.9 versus 23.4 pg/ml, p<0.01) than pre-anastrozole group, resulting in a decreased testosterone to estradiol ratio ($0.21{\pm}0.07$ versus $0.39{\pm}0.15$, p<0.01). Semen analyses before and during anastrozole treatment revealed significant increases in sperm count (35.5 versus 52.2 million sperm per ml, p<0.01) and motility (22.9% versus 29.3%, p<0.01). Conclusions: We identified infertile men with testicular failure had hormonal changes characterized by a decreased serum testosterone to estradiol ratio. The ratio can be corrected with aromatase inhibitor, resulting in a significant improvement in semen parameters.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.3
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• pp.439-447
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• 1992

Eight healthy Holstein bulls, 4-6 years old were used to study the effect of season of the year on the biophysical characteristics of semen. Semen was collected twice a week by AV (artificial vagina) over one-year period. The analyses revealed that all the basic seminal traits studied were differed significantly due to season, except the ejaculate volume and consistency and the percentage of swollen spermatozoa in a hypo-osmotic fructose-citrate solution. Ejaculates collected during hot summer season had significantly lower sperm motility, concentration and total counts, and higher percentage of dead spermatozoa than those collected during winter time. Warm spring had moderate semen quality. The temperature-humidity index was calculated and it was associated (p < 0.01) negatively with the ejaculate pH, sperm concentration and total counts, and positively with the % of dead sperms. Ejaculate volume, percentage of swollen spermatozoa, individual motilities did not correlate significantly with the change in temperature-humidity index values. The total live, motile spermatozoa per ejaculate during both the winter and spring seasons showed significant increase of about 37% and 32% respectively over the summer season. Also, rectal temperatures of the bulls were elevated during the hot summer season, while the values of blood hemoglobin and packed-cell volume were decreased.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.3
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• pp.159-166
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• 2007

Objective: To investigate whether semen parameters in infertile couples who undergone intrauterine insemination (IUI) change in the subsequent IUI cycle and the subsequent in vitro fertilization (IVF) cycle. Methods: Fifty-three infertile couples who had failed to become pregnant after the first IUI cycle with computer-assisted semen analysis (CASA) were included. After the first IUI, thirty-eight couples underwent the second IUI (Group 1), and fifteen underwent IVF-ET procedure (Group 2). All semen parameters including semen volume, concentration, motility and total motile sperm count were analyzed in the second IUI or IVF-ET procedure for comparison with the result of first IUI. Results: There were no significant differences in husband age, interval between the first and second procedure and cause of infertility. In Group 1, only sperm motility at the time of the latter IUI was significantly decreased when compared to the former IUI irrespective of the first semen parameters. In Group 2, sperm concentration, motility and total motile sperm count at the time of subsequent IVF were lower than the former IUI. By sub-analyses of Group 2, in the group of optimal semen parameter at IUI cycle, sperm concentration and total motile sperm count at the time of subsequent IVF were lower than the former IUI, while in the group of suboptimal semen parameter at IUI cycle, only sperm motility at the time of subsequent IVF were lower than the former IUI. Conclusion: The semen parameters in couples converted to IVF cycle were more adversely affected than those remained in IUI cycle. Further study on psychological stress should be necessary to explain the reason.