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Sperm Analysis (sperm + analysis)
Selected AbstractsSome characteristics of sperm motility in European hake (Merluccius merluccius, L., 1758)JOURNAL OF APPLIED ICHTHYOLOGY, Issue 5 2010A.-L. Groison Summary The objective of this paper is to characterize some of the sperm motility parameters in European hake (Merluccius merluccius), which is considered to be a species with aquaculture potential. The total ATP, ADP and AMP concentrations were determined using high-performance liquid chromatography on hake sperm samples collected during the winter-early spring in the Bay of Biscay (France) (n = 22) and on hake sperm samples collected during the summer-early autumn in waters off Western Norway (n = 5). The Adenylate Energy Charge (AEC) was deduced from these data. Computer Assisted Sperm Analysis (CASA) was used to measure a series of parameters characterizing the motility and the sperm swimming performances. Changes in salinity of the swimming medium affected all the measured motility parameters. The sperm velocity and the straightness of the movement were at maximum when sperm was activated with 100% filtrated sea water (100 SW) but decreased sharply later. When sperm was activated in filtrated sea water (50% diluted with distilled water: 50 SW) the values of these parameters increased (with a lower percentage of active cells) during the first 2.5 min and thereafter decreased slowly. In 50 SW, the initial velocity was lowered but the swimming period lasting 4.5 times longer than in 100 SW (but with a lower percentage of actively swimming cells). Initial sperm motility (percentage of swimming cells) in 100 SW was affected by sperm storage duration. Undiluted sperm could be stored at 4°C for 5 days and still show 13 ± 7% motility; the velocity and straightness of the movement were at maximum at the earliest period of measurement (0.5,1 day of storage) and then decreased gradually to reach their minima after 4 days of storage. Further, both the AEC and ATP content decreased with storage time, with the AEC decreasing from 0.78 ± 0.07 (mean ± SD) at stripping time to 0.20 ± 0.09 after 2 days of storage. Over the same period ATP content decreased from 85 ± 80 to 5 ± 4 nanomoles 10,9 spermatozoa, these data presenting a high variability. [source] Combined repeated dose and reproductive/developmental toxicity screening test of the nitrophenolic herbicide dinoseb, 2- sec -butyl-4,6-dinitrophenol, in ratsENVIRONMENTAL TOXICOLOGY, Issue 2 2008Mariko Matsumoto Abstract In a combined repeated dose toxicity study with reproduction/developmental toxicity screening test, Crj:CD(SD)IGS rats were dosed with dinoseb, 2- sec -butyl-4,6-dinitrophenol, by gavage at 0 (vehicle), 0.78, 2.33, or 7.0 mg/kg bw/day. Six males per group were dosed for a total of 42 days beginning 14 days before mating. Twelve females per group were dosed for a total of 44,48 days beginning 14 days before mating to day 6 of lactation throughout the mating and gestation period. Recovery groups of six males per group and nonpregnant six females per group were dosed for 42 days followed by a 14-day recovery period. No deaths were observed in males of any dose group or in females of the recovery groups. At 7.0 mg/kg bw/day, eight females died and two animals were moribund during late pregnancy, and a significant decrease in body weight gain was found in both sexes. Hematocrit was significantly higher at 0.78 mg/kg bw/day and above in the main group males at the end of administration period. Reduction in extramedullary hematopoiesis in the spleen was significant at 2.33 mg/kg bw/day in the main group females. Sperm analysis revealed a decrease in sperm motility and an increase in the rates of abnormal sperm, abnormal tail, and abnormal head at 7.0 mg/kg bw/day. A number of dams delivered their pups and of dams with live pups at delivery was significantly lowered in the 7.0 mg/kg bw/day group. Based on these findings, the LOAEL for males and NOAEL for females were 0.78 mg/kg bw/day, and the NOAEL for reproductive/developmental toxicity was considered to be 2.33 mg/kg bw/day. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2008. [source] Expression of aquaporins in the efferent ductules, sperm counts, and sperm motility in estrogen receptor-, deficient mice fed lab chow versus caseinMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 2 2006Ricardo Ruz Abstract Estrogens play an important role in the male reproductive tract, and this is especially so for the efferent ductules, where ,-estrogen receptors (ER,) have been localized. Mice deficient in ER, (,ERKO mice) are infertile, and the effect appears to be due in part to retention of water at the level of the efferent ductules. In the present study, we examined the consequences of ER, deletion on the distribution of certain aquaporins (AQPs), water protein channels, in the efferent ductules and on sperm numbers and motility. In addition, the effects of feeding mice a regular lab chow diet, which contains phytoestrogens, known to affect male reproductive tract functions, and a casein diet, which lacks phytoestrogens, were also assessed. Light microscope immunolocalizations of AQP-1 and AQP-9 revealed dramatic reduction and patchier staining in ,ERKO mice with distal areas of the efferent ductules being more affected than proximal areas. No other changes in immunolocalizations were noted as a consequence of diet. Computer-assisted sperm analyses demonstrated a 62% reduction in cauda epididymal sperm/ml in ,ERKO mice fed lab chow, whereas 87% fewer sperm/ml were observed in ,ERKO mice fed casein, suggesting an enhanced role for sperm production and concentration in a diet containing phytoestrogens. All sperm motility parameters were altered to some degree in ,ERKO mice fed lab chow. Alterations in sperm motility parameters were also detected, but were less dramatic in ,ERKO mice fed casein. These data suggest that the decrease in AQP expression in the efferent ductules of ,ERKO mice contributes in part to water retention in this tissue, eventually leading to backflow of water into the testis, with subsequent decreases in sperm concentration and motility. The data also suggest that phytoestrogens, which are present in regular lab chow, can influence the male reproductive tract with and without the presence of ER,, promoting efferent ductule and epididymal functions when ER, is expressed, but inhibiting these same functions when ER, is missing. Taken together the data underscore the importance of estrogens and ER, in maintaining sperm maturation and preventing male infertility. Mol. Reprod. Dev. © 2005 Wiley-Liss, Inc. [source] OC1 Effects of Cryopreservation on the Expression of Glut-3, Glut-5 and As-A Proteins in Iberian Boar Sperm MembranesREPRODUCTION IN DOMESTIC ANIMALS, Issue 2006S Sancho In order to determine the injure produced in boar spermatozoa through cryopreservation process, we analyzed the expression of the hexose transporters Glut-3 and Glut-5 and the zona pellucida binding protein As-A (P68) in three different steps of the freezing-thawed protocol: at 17°C (fresh BTS-diluted semen, 1 : 2 v/v, step 1), at 5°C (after glycerol addition; step 2), and post-thawing (step 3). All sperm analyses were carried out with immunogold techniques under electronic microscopy. For this study eight healthy post-pubertal Iberian boars were submitted to a collection of twice per week through 3 months, evaluating two ejaculates from each boar. Glut-3 maintains the expression in the acrosome region post-thawing but not along the tail where is reduced. The expression of Glut-5 and As-A is majority located at the post-acrosome region of the spermatozoa at step 1, but in step 2 and step 3 this expression is relocated to sperm tail area. In conclusion, while cryopreservation affects the localization and the expression of Glut-3 and Glut-5, its fertilizing capacity is not significantly reduced. The stabilization of boar semen at 5°C was found to be the most crucial step for sperm survival. [source] Integrity of mitochondrial membrane potential reflects human sperm qualityANDROLOGIA, Issue 1 2009J. A. Espinoza Summary The aim of this work was to evaluate intracellular reactive oxygen species (ROS) levels, phosphatidylserine (PS) externalisation and mitochondrial membrane potential integrity in the spermatozoa of healthy donors and outpatients who consulted for infertility and to correlate the results with the classic sperm parameters. For the evaluation of intracellular ROS levels, PS externalisation and mitochondrial membrane potential integrity, the fluorescent compounds dihydroethidium, annexin V-FITC and JC-1, respectively, were used and analysed by using flow cytometry. Conventional seminal analysis, including motility, viability, morphology, sperm count and volume, was performed according to the WHO criteria. The mitochondrial membrane potential and ROS results showed significant differences between the spermatozoa of individuals with a normal semen analysis and those of the group presenting abnormality in at least one of the sperm parameters. Mitochondrial membrane potential showed a significant and direct correlation with all the sperm parameters analysed. ROS were inversely correlated with motility, viability and morphology. PS externalisation, however, did not show any differences between the two groups, nor was it correlated with the sperm parameters examined. The evaluation of mitochondrial membrane potential integrity is a test that reflects sperm quality, which makes it highly recommendable to be applied as a complement to routine sperm analyses. [source] Automatic annular laser trapping: a system for high-throughput sperm analysis and sortingJOURNAL OF BIOPHOTONICS, Issue 3 2009Linda Shi Abstract An automatic microscope system is designed to study the response of sperm motility to an annular laser trap. A continuous annular laser trap provides a parallel way to analyze and sort sperm based on their motility and to study the effects of laser radiation, optical force and external obstacles. In the described automatic microscope system, the phase contrast images of swimming sperm are digitized to the computer at video rates. The microscope stage is controlled in real-time to relocate the sperm of interest to the annular trap with a normal or tangential entering angle. The sperm is continuously tracked and the swimming behavior is identified. Using this system, parallel sorting on human and gorilla sperm are achieved and threshold power levels separating the "fast" group and the "slow" group are compared for those two species. (© 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Sperm defects in mice lacking a functional Niemann,Pick C1 protein,MOLECULAR REPRODUCTION & DEVELOPMENT, Issue 10 2006Jun Fan Abstract The Niemann,Pick C1 (NPC1) gene encodes for a multiple membrane spanning protein, which regulates the trafficking of low-density lipoprotein-mediated endocytosed cholesterol. Mutation of the human NPC1 gene causes Niemann,Pick type C (NPC) disease. The Npc1NIH mice, a model of human NPC disease, bear a spontaneous mutation of the Npc1 gene, and are infertile. In this study, we have performed sperm analysis to search for the cause of male infertility in the Npc1NIH mouse. The number of cauda sperms in Npc1,/, mice was decreased roughly three-and-half-fold of that in wild-type mice. The decreased sperm number in Npc1,/, mice is due, at least in part, to partial arrest of spermatogenesis in the testes, as revealed by histological analysis. Compared to wild-type sperm, Npc1,/, sperm displayed a high frequency of morphological abnormalities, including tailless heads and aberrant heads. In the in vitro fertilization (IVF) assay using cumulus-intact eggs, Npc1,/, sperm failed to produce two-cell embryos. In the IVF assay where zona-free eggs were used, Npc1,/, sperm bound normally but could not fuse with the egg. Further analysis indicated that Npc1,/, sperms are drastically impaired in the binding to the egg zona pellucida, only 14% of the level of wild-type sperm. Moreover, on Npc1,/, cauda sperm, one-third of the total cyritestin protein was not proteolytically processed, while fertilin , was processed normally. Taken together, these results demonstrate that there are multiple defects in sperms from mice lacking a functional NPC1 protein, and these observed sperm defects may result in sterility. Mol. Reprod. Dev. © 2006 Wiley-Liss, Inc. [source] Effects of FSH receptor deletion on epididymal tubules and sperm morphology, numbers, and motilityMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 2 2005Amit Grover Abstract Follicle stimulating hormone (FSH) interacts with its cognate receptor (R) on Sertoli cells within the testis and plays an important role in the maintenance of spermatogenesis. Male FSH-R knockout (FORKO) mice show fewer Sertoli cells and many that are structurally abnormal and as a consequence fewer germ cells. Lower levels of serum testosterone (T) and androgen binding protein (ABP) also occur, along with reduced fertility. To assess the effects of FSH-R depletion as an outcome of testicular abnormalities, sperm from the cauda epididymidis were counted and examined ultrastructurally. As reduced fertility may also reflect changes to the epididymis, the secondary responses of the epididymis to lower T and ABP levels were also examined by comparing differences in sizes of epididymal tubules in various regions of FORKO and wild type (WT) mice. Sperm motility was evaluated in FORKO mice and compared to that of WT mice by computer assisted sperm analysis (CASA). Quantitatively, the data revealed that epithelial areas of the caput and corpus epididymidis were significantly smaller in FORKO mice compared to WT mice. Cauda epididymal sperm counts in FORKO mice were also much lower than in WT mice. This resulted in changes to 9 out of 14 sperm motility parameters, related mostly to velocity measures, which were significantly lower in the FORKO mice. The greatest change was observed relative to the percent static sperm, which was elevated by 20% in FORKO mice compared to controls. EM analyses revealed major changes to the structure of the heads and tails of cauda luminal sperm in FORKO mice. Taken together these data suggest a key role for the FSH receptor in maintaining Sertoli cells to sustain normal sperm numbers and proper shapes of their heads and tails. In addition, the shrinkage in epididymal epithelial areas observed in FORKO mice likely reflect direct and/or indirect changes in the functions of these cells and their role in promoting sperm motility, which is noticeably altered in FORKO mice. Mol. Reprod. Dev. © 2005 Wiley-Liss, Inc. [source] Long-term ofloxacin testicular toxicity: an experimental studyANDROLOGIA, Issue 2 2010M. A. EL-Harouny Summary The aim of this study was to assess the long-term toxic effect of ofloxacin on the testes and epididymides of 72 adult male albino rats. The rats were divided into group A and group B. Group A, which received ofloxacin for 14 days, was subdivided into two subgroups; LD-14 received low dose 72 mg KBW,1 daily and HD-14 received high dose 216 mg KBW,1 daily. Group B, which received ofloxacin for 28 days, was subdivided into two subgroups; LD-28 received 72 mg KBW,1 and HD-28 received 216 mg KBW,1 daily. Two matched control groups were followed up for 14 and 28 days respectively. The animals were evaluated for body weight, testicular weight, relative testicular weight, serum testosterone (T), epididymal sperm analysis (sperm count, motility, morphology, curvilinear velocity, linear velocity and linearity index) and testicular histopathology. The adverse effects of ofloxacin were correlated with increased treatment duration and/or dose. It is concluded that long-term ofloxacin has a direct detrimental effect on the testicles of albino rats at the studied doses and durations. [source] C3 in seminal plasma has no additional informative value in the diagnosis of infection/inflammation of the male genital tractANDROLOGIA, Issue 2 2003R. Boit Summary. The objective of this study was to determine the clinical significance of complement fraction C3 (C3c) in seminal plasma. Therefore 120 samples from randomly chosen subfertile males without signs of genital tract infection were screened for C3 and for seminal leucocytes as markers for subclinical infection/inflammation. A comprehensive semen evaluation included sperm analysis, sperm migration testing, immunocytochemical round cell differentiation to determine seminal leucocyte counts and the leucocyte ratio and semen cultures, in aliquots of the same ejaculates. C3 concentrations were significantly correlated with leucocyte counts per ml (P < 0.002) and per ejaculate (P < 0.001), and with the leucocyte ratio (P < 0.001). No association of C3 concentrations with semen quality or with the bacterial colonization of semen samples was found. The significant association with seminal leucocytes suggests that C3 might be used as an additional marker for silent male genital tract infection. In comparison with semen leucocytes, C3 screening does not reveal any further information about semen quality or infection/inflammation pathogenesis of the male genital tract. [source] Effect of varicocelectomy on seminal plasma transferrin values: a comparative clinical trialANDROLOGIA, Issue 1 2000A. Kosar Summary. The possible effects of varicocele and of the varicocelectomy procedure on Sertoli cell function were investigated. Transferrin concentrations in seminal plasma in men with varicocele before and 3 months after the operation were evaluated. Concentrations were measured in 10 normozoospermic fertile men as a control group and 32 oligozoospermic men with varicocele. Also, sperm analysis before and 3 months after the operation was performed. The mean transferrin level in seminal plasma was 108.4 ± 17.5 ,g, ml,1 in normoozoospermic men and 58.1 ± 14.4 ,g ml,1 in patients with varicocele before the operation (P < 0.0001). Mean sperm concentration, motility and normal morphology ratio showed significant improvement 3 months after the operation (P < 0.0001). Although the mean transferrin level increased slightly (to 60.8 ± 16.2 , ml,1; P=0.2), there was a statistically significant correlation between the change in transferrin concentration and the change in sperm concentration after the operation (r=0.56, P=0.0008). These results showed that elevated transferrin secretion after the treatment seems to be associated with an increase in sperm concentration after varicocelectomy. The finding of improvements in seminal parameters after the operation but insignificant changes in seminal transferrin levels indicates that varicocelectomy results in a greater improvement in sperm quality than in Sertoli cell function., [source] Effect of different methods for the induction of spermiation on semen quality in European eelAQUACULTURE RESEARCH, Issue 15 2005Juan F Asturiano Abstract Five hormonal treatments with human chorionic gonadotropin (hCG) were tested for the induction of maturation and spermiation in male farmed eels. The main aim was to optimize previously used hormonal treatments to achieve shorter induction treatments, longer spermiation periods and/or higher sperm quality. Fish treated for just 3 weeks (treatment E) or until the onset of spermiation (treatment C) showed the worst results, while the treatment consisting of weekly administration of 1.5 IU hCG g,1 fish (treatment A) induced the highest percentage of spermiating males, the highest number of sperm samples and sperm volumes and densities similar to the rest of the treatments (B: half hormone dosage, or D: biweekly administration). Evaluation of the sperm quality was performed by computer-assisted sperm analysis (CASA), considering the percentage of total motile spermatozoa, the percentage of fast and medium-velocity spermatozoa, as well as different motility parameters. Sperm samples from A-D groups showed between 44% and 54% motile spermatozoa, and between 10% and 15% fast spermatozoa, while samples from E-treated males showed 0% motile cells. No significant differences were found in the spermatozoa straight line velocity (VSL), curvilinear velocity (VCL) or the angular velocity (VAP), neither spermatozoa beating cross frequency (BCF) between A,D groups. [source] | |||||||||||||