The very low cut-off value for sperm morphology of 4% morphologically normal spermatozoa, as proposed in the new edition of the World Health Organization （WHO） manual on semen analysis, is in agreement with recently...The very low cut-off value for sperm morphology of 4% morphologically normal spermatozoa, as proposed in the new edition of the World Health Organization （WHO） manual on semen analysis, is in agreement with recently published values and reflects the trend of a decline in reported mean values for normal sperm morphology. The reduced value for morphologically normal spermatozoa over the years may be due to several factors. The first is the introduction of strict criteria for the evaluation of sperm morphology. Other reasons may include the introduction of additional criteria for sperm morphology abnormalities and the suggested decrease in semen parameters because of increasing negative environmental influences. Although on its own the newly proposed very low normal value may not provide the strong predictive value for a males＇ fertility potential, as originally reported for sperm morphology evaluated according to strict criteria, a good predictive value can still be obtained if the holistic, strict approach for sperm morphology evaluation is followed together with additional sperm morphology parameters now available, because certain morphology patterns and sperm abnormalities are now known to be of strong prognostic value. In addition, better international standardization of the technical methodology, consensus on the interpretation of sperm morphology evaluation criteria and standardized international external quality control （EQC） schemes, are of utmost importance to maintain the good predictive value of sperm morphology.展开更多
The measurement or evaluation and clinical significance of human sperm morphology has always been and still is a controversial aspect of the semen analysis for the determination of a male＇s fertility potential. In th...The measurement or evaluation and clinical significance of human sperm morphology has always been and still is a controversial aspect of the semen analysis for the determination of a male＇s fertility potential. In this review the background of the development of the evaluation criteria for sperm morphology will be discussed. Aspects of criticism on the strict criteria definition and use of the criteria for sperm morphology evaluation will be discussed as well as possible reasons for the decline in normal sperm morphology values and how we can compromise for this phenomenon resulting in the very low normal reference value as published in the 2010 WHO manual for the Examination and Processing of Human Semen. One of the possible solutions may be to give more attention to a limited number of abnormal sperm morphology categories and the inclusion of sperm morphology patterns. It is concluded in this review that if done correctly and with care and with strict application of existing guidelines as outlined in the 2010 WHO manual, sperm morphology measurement still has a very important role to play in the clinical evaluation of male fertility potential.展开更多
The new World Health Organization （WHO） Manual for Semen Analysis contains several improvements. One is that the 20 million spermatozoa per mL paradigm has been ousted in favour of proper calculations of lower refer...The new World Health Organization （WHO） Manual for Semen Analysis contains several improvements. One is that the 20 million spermatozoa per mL paradigm has been ousted in favour of proper calculations of lower reference limits for semen from men, whose partners had a time-to-pregnancy of 12 months or less. The recommendation to grade the progressive motility as described in the third and fourth editions of the WHO manual was not evidence-based, and WHO was therefore motivated to abandon it. However, the new recommendation is not evidence-based either, and it is difficult to understand the rational for the new assessment. It may have been a compromise to avoid returning to the rather robust system recommended in the first edition （1980）. The unconditional recommendation of the ＇Tygerberg strict criteria＇ is not evidence-based, and seems to be the result of an unfortunate bias in the composition of the Committee in favour of individuals known to support the ＇strict criteria＇ method. This recommendation will have negative effects on the develop- ment ofandrology as a scientific field. Given the importance of the WHO manual, it is unfortunate that the recommenda- tions for such important variables, as motility and morphology, lack evidence-based support.展开更多
To provide basic toxicity data for formulating risk characterization benchmarks, the effects of lead on survival, locomotion, and sperm morphology were investigated in the Asian earthworm Pheretima guillelmi. The LC50...To provide basic toxicity data for formulating risk characterization benchmarks, the effects of lead on survival, locomotion, and sperm morphology were investigated in the Asian earthworm Pheretima guillelmi. The LC50 of P. guiUelmi for 7 and 14 d were 4285＋339 mg/kg and 3207＋248 mg/kg, which shows P. guillelmi can tolerate a higher concentration of lead nitrate. The average weight of the surviving earthworms decreased at concentration of 2800 mg Pb/kg soil, and the locomotor ability of earthworms exposed to a range of soil Pb concentrations showed a general decrease with increasing Pb concentrations. We also presented data depicting the sperm morphology of earthworms, which shows potential as a sensitive biomarker for measuring the effects of heavy metal on reproduction.展开更多
For infertility treatment,the selection of in vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)is decided by multiplying indicators(including fallopian tube factors,semen count,and semen motility),exce...For infertility treatment,the selection of in vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)is decided by multiplying indicators(including fallopian tube factors,semen count,and semen motility),except for sperm morphology.In this study,we conducted a retrospective analysis,from implantation to birth,over a period of 5 years.A total of 1873 couples with primary or secondary fallopian tube factors and an increased defective sperm morphology rate(DSMR)were divided into different groups to receive IVF or ICSI cycles.By comparing the outcomes,we found that the F1 group(DSMR<96%,IVF group 1)had higher cleavage rate,biochemical pregnancy rate,clinical pregnancy rate,and live birth rate than the F3 group(DSMR>98%,IVF group 3;P<0.05).In contrast,there was no significant difference in the ICSI subgroups.Furthermore,a comparison of the outcomes between IVF and ICSI showed that the S3 group(DSMR>98%,ICSI group 3)had higher cleavage rate(P<0.001),biochemical pregnancy rate(P<0.05),clinical pregnancy rate(P<0.05)and live birth rate(P<0.05)than the F3 group.However,the ICSI subgroup had a lower two pronuclei fertilization rate than the IVF subgroup(P<0.05).Our data suggest that the sperm morphology should also be considered when selecting IVF or ICSI combined with other semen parameters before the first assisted reproductive technologies(ART)cycle,especially for males with severe sperm defects.展开更多
Aberrant sperm protamination is linked to sperm dysmorphology and nuclear chromatin condensation.Yet,its effects on sperm cytoplasmic maturation remain largely unexplored.The relationships of protamines,sperm morpholo...Aberrant sperm protamination is linked to sperm dysmorphology and nuclear chromatin condensation.Yet,its effects on sperm cytoplasmic maturation remain largely unexplored.The relationships of protamines,sperm morphology,DNA damage,and cytoplasmic remodeling were illustrated in this study to provide fresh perspectives on the mechanisms of male infertility.A total of 205 infertile males were allocated into 5 groups according to the percentage of spermatozoa exhibiting abnormal morphology within their samples.Sperm concentration,motility,abnormal sperm morphology,cytoplasmic droplets(CDs),and excess residual cytoplasm(ERC)were analyzed according to the World Health Organization manual(2010).Sperm nuclear vacuoles(NVs)were determined by propidium iodide(PI)staining.Sperm protamine expressions(P1 and P2)were detected by western blot.DNA damage was measured by acridine orange test(AOT)to calculate the proportion of sperm with single-strand DNA breaks(SSBs).Our data showed that sperm concentration and motility in infertile males significantly decreased with the severity of abnormal sperm morphology(both P<0.01).P1 level,P1/P2 ratio,and SSB rate increased with the severity of sperm dysmorphology,whilst the P2 level decreased(all P<O.01).NVs,CDs,and ERC were more common in males with sperm dysmorphology and positively correlated with the SSB rate(all P<O.01).The relationships between the SSB rate and the P1/P2 ratio were also significant(P<0.01).Aberrant protamination may cause sperm dysmorphology and compromise male fertility by impairing sperm's nucleus and cytoplasm maturation,with the P1/P2 ratio potentially serving as a valuable indicator of sperm quality and male fertility.展开更多
Background:Collecting and studying live sperm is central to many important fields of biology.Yet,a simple method to collect live sperm is lacking in wild seabird species.Here,we describe a non?invasive method to colle...Background:Collecting and studying live sperm is central to many important fields of biology.Yet,a simple method to collect live sperm is lacking in wild seabird species.Here,we describe a non?invasive method to collect viable sperm samples based on a simple massage technique applied to male Black?legged Kittiwakes(Rissa tridactyla).Methods:We studied a colony breeding at Kongsfjorden,Svalbard and successfully obtained sperm samples from 32 males.With a subset of samples(n = 12 males),we compared the suitability of several extenders(0.9% NaCl,PBS,Earle's balance salt solution,Dulbecco's modified Eagle medium) in maintaining sperm alive long enough for analyses.With another 18 ejaculates,we conducted computer assisted sperm analyses using the CASA plugin for ImageJ.We provide details about the settings to be used for such analyses.Lastly,droplets from 20 ejaculates were smeared on glass slides and preserved with formalin to characterize sperm morphology in terms of total sperm length,sperm head length,midpiece length and flagellum length,and percentage of abnormal sperm.Results:With this method and under field conditions,we were able to obtain sufficient amounts of live sperm to assess traits related to sperm quality(e.g.sperm morphology,percentage of motile sperm,sperm velocity).We found that two extenders,Earle's balanced salt solution and Dulbecco modified Eagle's medium,yielded similarly good results.Additionally,we investigated whether specific behaviours were associated with successful sperm collection and whether sperm collection success depended on how long before laying sperm collection was attempted.Finally,we provide mean values for sperm morphology,sperm swimming ability and percentage of motile sperm,which may prove useful for future comparative analyses,and we report high levels of sperm abnormality and within?ejaculate variation in sperm morphology.Conclusions:We discuss the high percentage of abnormal sperm and high within?ejaculate variation in sperm morphology in light of sperm competition theory and conclude that these figures are likely due to relaxed post?cop?ulatory sexual selection,kittiwakes being strictly monogamous.Finally,we suggest that this method could be applied to other seabird species sharing similar ecology.展开更多
The study investigated the intervention of caffeine on the effect of Nigerian Bonnylight crude oil (NBLCO) on sperm motility and morphology of diabetic Wistar rats. Eighty adult male rats (180 - 200 g body weight) wer...The study investigated the intervention of caffeine on the effect of Nigerian Bonnylight crude oil (NBLCO) on sperm motility and morphology of diabetic Wistar rats. Eighty adult male rats (180 - 200 g body weight) were randomly divided into eight groups of 10 animals each. The control group received 3 mL/Kg body weight of distilled water, ND Caf. group received 20 mL/kg body weight of caffeine, NDCO group received 3 mL/Kg body weight of NBLCO, ND Caf. + CO group received 20 and 3 mL/Kg body weight of caffeine and NBLCO respectively, diabetic group received 3 mL/Kg body weight of distilled water, D Caf. group received 20 mL/Kg body weight of caffeine, D CO received 3 mL/Kg body weight of NBLCO and D Caf. + CO group received 20 and 3 mL/Kg body weight of caffeine and NBLCO respectively, by oral gavaging for 28 days. The results showed that independent administration of caffeine and NBLCO to both non-diabetic and diabetic rats significantly (p < 0.05) altered sperm concentration, morphology and motility. Administration of NBLCO aggravated the worsening condition by significantly (p < 0.05) reducing sperm concentration, motility and increasing abnormal sperms in diabetic rats. Although caffeine significantly (p < 0.05) reduced motility in non-diabetic rats, it did cause any significant alteration in diabetic rats. Diabetes mellitus and NBLCO significantly (p < 0.05) reduced all indices that promote sperm motility while significantly (p < 0.05) increasing indices that do not promote motility. It can be concluded that diabetes mellitus and NBLCO administration have shown the tendency to promote male infertility by reducing sperm motility and adversely altering the morphology of sperm in male Wistar rats, which was ameliorated by caffeine intervention.展开更多
In this study, we compared conventional sperm selection with high-magnification morphology based on the motile sperm organellar morphology examination （MSOME） criteria, and hyaluronic acid （HA） binding for sperm c...In this study, we compared conventional sperm selection with high-magnification morphology based on the motile sperm organellar morphology examination （MSOME） criteria, and hyaluronic acid （HA） binding for sperm chromosome aneuploidy and DNA fragmentation rates. Semen from 50 severe male factor cases was processed through density gradient centrifugation, and subjected to sperm selection by using the conventional method （control）, high magnification at x6650 or HA binding. Aneuploidy was detected by fluorescence in situ hybridization with probes for chromosomes 13, 18, 21, X and Y, and DNA fragmentation by the terminal deoxynucleotidyl transferase dUTP nick end labelling （TUNEL） method. Spermatozoa selected under high-magnification had a lower DNA fragmentation rate （2.6% vs. 1.7%; P=0.032）, with no significant difference in aneuploidy rate （0.8% vs0.7%; P=0.583）, than those selected by the HA binding method. Spermatozoa selected by both methods had much lower aneuploidy and DNA fragmentation rate than the controls （7% aneuploidy and 26.8% DNA fragmentation rates, respectively）. In the high-magnification group, the aneuploidy rate was lower when the best spermatozoa were selected than when only the second-best spermatozoa were available for selection, but the DNA fragmentation rate was not different. In conclusion, sperm selection under high magnification was more effective than under HA binding in selecting spermatozoa with low DNA fragmentation rate, but the small difference （0.9%） might not be clinically meaningful. Both methods were better than the conventional method of sperm selection.展开更多
Multiple morphological abnormalities of sperm flagella(MMAF)is a severe form of asthenoteratozoospermia,characterized by morphological abnormalities and reduced motility of sperm,causing male infertility.Although appr...Multiple morphological abnormalities of sperm flagella(MMAF)is a severe form of asthenoteratozoospermia,characterized by morphological abnormalities and reduced motility of sperm,causing male infertility.Although approximately 60%of MMAF cases can be explained genetically,the etiology of the remaining cases is unclear.Here,we identified two novel compound heterozygous variants in the gene,dynein axonemal heavy chain 10(DNAH10),in three patients from two unrelated Pakistani families using whole-exome sequencing(WES),including one compound heterozygous mutation(DNAH10:c.9409C>A[p.P3137T];c.12946G>C[p.D4316H])in family 1 and another compound heterozygous mutation(DNAH10:c.8849G>A[p.G2950D];c.11509C>T[p.R3687W])in family 2.All the identified variants are absent or rare in public genome databases and are predicted to have deleterious effects according to multiple bioinformatic tools.Sanger sequencing revealed that these variants follow an autosomal recessive mode of inheritance.Hematoxylin and eosin(H&E)staining revealed MMAF,including sperm head abnormalities,in the patients.In addition,immunofluorescence staining revealed loss of DNAH10 protein signals along sperm flagella.These findings broaden the spectrum of DNAH10 variants and expand understanding of the genetic basis of male infertility associated with the MMAF phenotype.展开更多
Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both...Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both developing and developed countries. Other optional tests or sophisticated technologies have not been widely applied. The current review addresses important changes in the analysis of semen as described in the new World Health Organization （WHO） manual for semen analysis. The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality. Apart from the above mentioned changes, the initial evaluation and handling methods remain, in most instances, the same as in previous editions. Furthermore, the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology. WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995. The external qualitY control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training. This review reports on current sperm functional tests, such as the induced acrosome reaction, and sperm-zona pellucida binding assays, as well as the impact of sperm quality in terms of DNA integrity, and the relationship of sperm function tests to sperm morphology.展开更多
Objective:To study the effect of mobile emitted radio frequency electromagnetic radiation(RF-EMR)on sperm parameters(sperm count,viability,motility,progressivity,and morphology)and ameliorative effect of pomegranate j...Objective:To study the effect of mobile emitted radio frequency electromagnetic radiation(RF-EMR)on sperm parameters(sperm count,viability,motility,progressivity,and morphology)and ameliorative effect of pomegranate juice in rats.Methods:Thirty male Sprague Dawley rats were divided into five groups(n=6 in each group).GroupⅠwas neither exposed to RF-EMR nor given pomegranate juice,groupⅡ,ⅢandⅣwere exposed to mobile emitted RF-EMR for 60 min/day for 90 days.After 90-day exposure to RF-EMR,groupⅢwas supplemented with pomegranate juice for 90 days(1 mL/day)and groupⅣwas allowed to recover for 90 days without supplementation of pomegranate juice.GroupⅤwas supplemented with pomegranate juice for 90 days without exposure to RF-EMR.At the end of intervention,epididymal sperm parameters(sperm count,viability,motility,progressivity,and morphology)were measured.Results:The microscopic examination of sperm parameters such as sperm count,sperm viability,sperm motility,progressivity were significantly decreased in groupⅡ(the mobile RF-EMR group)compared to groupⅠ(the control group)(P<0.05).In addition,sperm morphology was also significantly altered(abnormal)in groupⅡcompared to groupⅠ(P<0.05).However,the sperm parameters including sperm morphology were significantly altered in groupⅢ(mobile RF-EMR+pomegranate juice)compared to groupⅡ(P<0.05).The sperm parameters including sperm morphology were not significantly altered in groupⅣ(the mobile RF-EMR recovery group)compared to groupⅡ(P>0.05).The sperm parameters were non-significantly increased in group V(the pomegranate juice group)compared to groupⅠ(P>0.05).Conclusions:Mobile RF-EMR exposure reduces the sperm parameters,which,however,can be reversed by treatment with pomegranate juice,indicating that pomegranate juice can be used as a nutritional supplement to improve sperm quality.展开更多
Objective:To determine the relationship between teratozoospermia and sperm DNA fragmentation(SDF)in the human ejaculate.Methods:This retrospective study included 100 normozoospermic men as a control cohort(abnormal fo...Objective:To determine the relationship between teratozoospermia and sperm DNA fragmentation(SDF)in the human ejaculate.Methods:This retrospective study included 100 normozoospermic men as a control cohort(abnormal forms>14%),210 patients with a high level of abnormal forms(≤4%)and 65 patients presenting with a moderate level of abnormal forms(>4%to≤14%)based on the World Health Organization definitions.Sperm morphology was assessed using bright field microscopy.Sperm DNA fragmentation was assessed using the sperm chromatin dispersion assay.Non-parametric analyses were conducted to examine the relationship between abnormal sperm morphology and sperm DNA fragmentation;receiver operating characteristic(ROC)analyses were conducted to assess sensitivity and specificity of this relationship.Results:A correlation analysis revealed that the higher the proportion of abnormal spermatozoa in the ejaculate,the higher the level of SDF(Spearman's Rho=-0.230;P<0.001).Significant differences in the proportion of SDF were found when all cohorts were compared(P<0.001);these significant differences were also retained when the different cohorts were compared pairwise.ROC analysis showed a moderate but significant predictive value for SDF to differentiate patients with different levels of teratozoospemia.Conclusions:Although analysis of a more continuous range of values for teratozoospermia would help further clarify any causal relationship with SDF,there is clearly a synergistic or coincident affiliation between these variables that needs to be acknowledged by the clinician when interpreting the spermiogram.展开更多
Aim： To investigate the reversibility of the effect of nandrolone decanoate （ND） on sperm parameters after discontinuing the drug. Methods： Three groups of rats received peanut oil （control）, low and high doses ...Aim： To investigate the reversibility of the effect of nandrolone decanoate （ND） on sperm parameters after discontinuing the drug. Methods： Three groups of rats received peanut oil （control）, low and high doses of ND for 14 weeks, respectively. Each group was divided into subgroups A and B, in which rats were killed 14 and 28 weeks after initiating the injection, respectively. Results： Sperm count and motile sperm fraction were decreased in the subgroups A and B that received low and high doses of ND in comparison with the controls A and B, respectively. The sperm count and motile sperm fraction increased in the subgroups B that received low and high doses of ND in comparison with their corresponding subgroups A. The number of normal morphology sperm was decreased significantly in subgroups A receiving low and high doses of ND in comparison with the control subgroup A. However, this parameter was not decreased in subgroups B receiving low and high doses in comparison with the control subgroup B. The normal sperm morphology did not show any significant differences in the subgroups B in comparison with their corresponding subgroups A. Conclusion： The 14-week injection of low and high doses of ND decreases sperm quality and quantity in rats. These parameters were improved after discontinuing ND, but not recovered completely even when they are left untreated for 14 weeks.展开更多
Objective To investigate the age-long claim by the locales that the food thickener, M. urens seed, has antispermatogenic, hence, antifertility effects in man. Methods Eight-week old male Albino rats were used as the m...Objective To investigate the age-long claim by the locales that the food thickener, M. urens seed, has antispermatogenic, hence, antifertility effects in man. Methods Eight-week old male Albino rats were used as the mammalian model for this study. They were assigned to four groups of 6 rats each and treatment with the ethanol extract was for a period of 14 d. The treatment regimes were 70 mg/kg, 140 mg/kg, 210 mg/kg and 0 mg/kg BW in groups A, B, C and D, respectively. Extracts were prepared by Soxhlet extraction using 80% ethanol as the extracting solvent. The stock solution was prepared by dissolving 1 g of the paste extract in 10 ml corn oil （vehicle） to make up 100 mg/ml concentration. At the end of the treatment, sperm from the distal caudal epididymis was collected and analyzed for sperm count, sperm motility and sperm morphology. Results Significant reduction was observed in sperm count and sperm motility （P〈0.05）. The mean sperm count for group A was 6.27±0.02×10^6, for group B was 6.16±0.02×10^6 and group C had 6.0±0.0×10^6 sperm cells The control （group D） had a mean sperm count of 6.50±0.09×10^6 which was higher than that of any treated group. Results of the sperm motility test gave the following mean rates for motile sperm cells after treatment： group A, 57.6±% 2.1; group B, 50.0±4.0; group C, 45.0±4.0. The control had the highest mean motility rate of 72.3±2.1. The observed sperm abnormalities included unusual head with large acrosome, looped tailpiece, mid piece with distal droplet, pin head, pyriform head and long hook.Conclusion The anti-spermatogenic effects of the extract on the sperm in the Albino rat may lead to reduction of fertility.展开更多
Objective To evaluate the effect of the accessory gential gland on the male fertility. Methods The relevant seminal parameters of 32 normal donors and 35 patients with olioasthenospermia were measured by seminal routi...Objective To evaluate the effect of the accessory gential gland on the male fertility. Methods The relevant seminal parameters of 32 normal donors and 35 patients with olioasthenospermia were measured by seminal routine analysis, sperm morphology and accessory, gentital glands markers.Results The percentage of normal spermatozoa, the function of sperm penetration through cervical mucus and the enzyme activity of neutral α-1,4-glucosidase significantly declined as compared with the control. There was a significant negative relationship between the semen volume and the sperm motility for the oligoasthenospermia （r= -0.415, P〈0.05）, and a significant positive relationship between the sperm count and the percentage of normal sperm （r=0.393, P〈0.05）.Conclusion The abnormal function of epididymis exists in patients with oligoasthenospermia. Sperm function,for the oligoasthenospermia declines with an increase of the percentage of the abnormal sperm. The poorer the spermatogenises function of the testis is, the higher the percentage of the abnormal sperm appears.展开更多
Objective To introduce a rapid simple staining method for sperm morphology.Methods Liquifiedsemen was on the glass, fixed by methanol, treated with phosphonic acid buffer, stained by A, B staining solution separately,...Objective To introduce a rapid simple staining method for sperm morphology.Methods Liquifiedsemen was on the glass, fixed by methanol, treated with phosphonic acid buffer, stained by A, B staining solution separately, and rinsed by running water. Results After staining, the whole sper-matozoid was very clear. The cephalic zone of spermatozoid head , acrosome area , was colored pink; its caudal zone colored violet red or violet blue. The body and tail of spermatozoid colored pink or light blue. Normal and abnormal morphology of spermatozoid could be differentiated easily. The sloughed spermatogenic cells and white blood cells could be differentiated also. Conclusion This rapid staining method of semen smear can produce the same effect of other usual staining method , but the staining time is shorter and the procedure is simpler.展开更多
The syndrome of multiple morphological abnormalities of the sperm flagella(MMAF)is a specific kind of asthenoteratozoospermia with a mosaic of flagellar morphological abnormalities(absent,short,bent,coiled,and irregul...The syndrome of multiple morphological abnormalities of the sperm flagella(MMAF)is a specific kind of asthenoteratozoospermia with a mosaic of flagellar morphological abnormalities(absent,short,bent,coiled,and irregular flagella).MMAF was proposed in 2014 and has attracted increasing attention;however,it has not been clearly understood.In this review,we elucidate the definition of MMAF from a systematical view,the difference between MMAF and other conditions with asthenoteratozoospermia or asthenozoospermia(such as primary mitochondrial sheath defects and primary ciliary dyskinesia),the knowledge regarding its etiological mechanism and related genetic findings,and the clinical significance of MMAF for intracytoplasmic sperm injection and genetic coun sell ng.This review provides the basic kno wledge for MMAF and puts forward some suggestions for further investigations.展开更多
Numerous genes have been associated with multiple morphological abnormalities of the sperm flagella(MMAF),which cause severe asthenozoospermia and lead to male infertility,while the causes of approximately 50%of MMAF ...Numerous genes have been associated with multiple morphological abnormalities of the sperm flagella(MMAF),which cause severe asthenozoospermia and lead to male infertility,while the causes of approximately 50%of MMAF cases remain unclear.To reveal the genetic causes of MMAF in an infertile patient,whole-exome sequencing was performed to screen for pathogenic genes,and electron microscope was used to reveal the sperm flagellar ultrastructure.A novel heterozygous missense mutation in the outer dense fiber protein 2(ODF2)gene was detected,which was inherited from the patient’s mother and predicted to be potentially damaging.Transmission electron microscopy revealed that the outer dense fibers were defective in the patient’s sperm tail,which was similar to that of the reported heterozygous Odf2 mutation mouse.Immunostaining of ODF2 showed severe ODF2 expression defects in the patient’s sperm.Therefore,it was concluded that the heterozygous mutation in ODF2 caused MMAF in this case.To evaluate the possibility of assisted reproductive technology(ART)treatment for this patient,intracytoplasmic sperm injection(ICSI)was performed,with the help of a hypo-osmotic swelling test and laser-assisted immotile sperm selection(LAISS)for available sperm screening,and artificial oocyte activation with ionomycin was applied to improve the fertilization rate.Four ICSI cycles were performed,and live birth was achieved in the LAISS-applied cycle,suggesting that LAISS would be valuable in ART treatment for MMAF.展开更多
Multiple morphological abnormalities of the sperm flagella(MMAF)is a specific type of asthenoteratozoospermia,presenting with multiple morphological anomalies in spermatozoa,such as absent,bent,coiled,short,or irregul...Multiple morphological abnormalities of the sperm flagella(MMAF)is a specific type of asthenoteratozoospermia,presenting with multiple morphological anomalies in spermatozoa,such as absent,bent,coiled,short,or irregular caliber flagella.Previous genetic studies revealed pathogenic mutations in genes encoding cilia and flagella-associated proteins(CFAPs;e.g.,CFAP43,CFAP44,CFAP65,CFAP69,CFAP70,and CFAP251)responsible for the MMAF phenotype in infertile men from different ethnic groups.However,none of them have been identified in infertile Pakistani males with MMAF.In the current study,two Pakistani families with MMAF patients were recruited.Whole-exome sequencing(WES)of patients and their parents was performed.WES analysis reflected novel biallelic loss-of-function mutations in CFAP43 in both families(Family 1:ENST00000357060.3,p.Arg300Lysfs*22 and p.Thr526Serfs*43 in a compound heterozygous state;Family 2:ENST00000357060.3,p.Thr526Serfs*43 in a homozygous state).Sanger sequencing further confirmed that these mutations were segregated recessively in the families with the MMAF phenotype.Semiquantitative reverse-transcriptase polymerase chain reaction(qRT-PCR)was carried out to detect the effect of the mutation on mRNA of the affected gene.Previous research demonstrated that biallelic loss-of-function mutations in CFAP43 accounted for the majority of all CFAP43-mutant MMAF patients.To the best of our knowledge,this is the first study to report CFAP43 biallelic loss-of-function mutations in a Pakistani population with the MMAF phenotype.This study will help researchers and clinicians to understand the genetic etiology of MMAF better.展开更多
文摘The very low cut-off value for sperm morphology of 4% morphologically normal spermatozoa, as proposed in the new edition of the World Health Organization （WHO） manual on semen analysis, is in agreement with recently published values and reflects the trend of a decline in reported mean values for normal sperm morphology. The reduced value for morphologically normal spermatozoa over the years may be due to several factors. The first is the introduction of strict criteria for the evaluation of sperm morphology. Other reasons may include the introduction of additional criteria for sperm morphology abnormalities and the suggested decrease in semen parameters because of increasing negative environmental influences. Although on its own the newly proposed very low normal value may not provide the strong predictive value for a males＇ fertility potential, as originally reported for sperm morphology evaluated according to strict criteria, a good predictive value can still be obtained if the holistic, strict approach for sperm morphology evaluation is followed together with additional sperm morphology parameters now available, because certain morphology patterns and sperm abnormalities are now known to be of strong prognostic value. In addition, better international standardization of the technical methodology, consensus on the interpretation of sperm morphology evaluation criteria and standardized international external quality control （EQC） schemes, are of utmost importance to maintain the good predictive value of sperm morphology.
文摘The measurement or evaluation and clinical significance of human sperm morphology has always been and still is a controversial aspect of the semen analysis for the determination of a male＇s fertility potential. In this review the background of the development of the evaluation criteria for sperm morphology will be discussed. Aspects of criticism on the strict criteria definition and use of the criteria for sperm morphology evaluation will be discussed as well as possible reasons for the decline in normal sperm morphology values and how we can compromise for this phenomenon resulting in the very low normal reference value as published in the 2010 WHO manual for the Examination and Processing of Human Semen. One of the possible solutions may be to give more attention to a limited number of abnormal sperm morphology categories and the inclusion of sperm morphology patterns. It is concluded in this review that if done correctly and with care and with strict application of existing guidelines as outlined in the 2010 WHO manual, sperm morphology measurement still has a very important role to play in the clinical evaluation of male fertility potential.
文摘The new World Health Organization （WHO） Manual for Semen Analysis contains several improvements. One is that the 20 million spermatozoa per mL paradigm has been ousted in favour of proper calculations of lower reference limits for semen from men, whose partners had a time-to-pregnancy of 12 months or less. The recommendation to grade the progressive motility as described in the third and fourth editions of the WHO manual was not evidence-based, and WHO was therefore motivated to abandon it. However, the new recommendation is not evidence-based either, and it is difficult to understand the rational for the new assessment. It may have been a compromise to avoid returning to the rather robust system recommended in the first edition （1980）. The unconditional recommendation of the ＇Tygerberg strict criteria＇ is not evidence-based, and seems to be the result of an unfortunate bias in the composition of the Committee in favour of individuals known to support the ＇strict criteria＇ method. This recommendation will have negative effects on the develop- ment ofandrology as a scientific field. Given the importance of the WHO manual, it is unfortunate that the recommenda- tions for such important variables, as motility and morphology, lack evidence-based support.
基金supported by the Natural Science Fund of Zhejiang Province (No. Y506255)
文摘To provide basic toxicity data for formulating risk characterization benchmarks, the effects of lead on survival, locomotion, and sperm morphology were investigated in the Asian earthworm Pheretima guillelmi. The LC50 of P. guiUelmi for 7 and 14 d were 4285＋339 mg/kg and 3207＋248 mg/kg, which shows P. guillelmi can tolerate a higher concentration of lead nitrate. The average weight of the surviving earthworms decreased at concentration of 2800 mg Pb/kg soil, and the locomotor ability of earthworms exposed to a range of soil Pb concentrations showed a general decrease with increasing Pb concentrations. We also presented data depicting the sperm morphology of earthworms, which shows potential as a sensitive biomarker for measuring the effects of heavy metal on reproduction.
基金This project was supported by the Shanghai Municipal Science and Technology Major Project(2017SHZDZX01)State Key Laboratory of Reproductive Medicine(SKLRM-K202002).
文摘For infertility treatment,the selection of in vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)is decided by multiplying indicators(including fallopian tube factors,semen count,and semen motility),except for sperm morphology.In this study,we conducted a retrospective analysis,from implantation to birth,over a period of 5 years.A total of 1873 couples with primary or secondary fallopian tube factors and an increased defective sperm morphology rate(DSMR)were divided into different groups to receive IVF or ICSI cycles.By comparing the outcomes,we found that the F1 group(DSMR<96%,IVF group 1)had higher cleavage rate,biochemical pregnancy rate,clinical pregnancy rate,and live birth rate than the F3 group(DSMR>98%,IVF group 3;P<0.05).In contrast,there was no significant difference in the ICSI subgroups.Furthermore,a comparison of the outcomes between IVF and ICSI showed that the S3 group(DSMR>98%,ICSI group 3)had higher cleavage rate(P<0.001),biochemical pregnancy rate(P<0.05),clinical pregnancy rate(P<0.05)and live birth rate(P<0.05)than the F3 group.However,the ICSI subgroup had a lower two pronuclei fertilization rate than the IVF subgroup(P<0.05).Our data suggest that the sperm morphology should also be considered when selecting IVF or ICSI combined with other semen parameters before the first assisted reproductive technologies(ART)cycle,especially for males with severe sperm defects.
文摘Aberrant sperm protamination is linked to sperm dysmorphology and nuclear chromatin condensation.Yet,its effects on sperm cytoplasmic maturation remain largely unexplored.The relationships of protamines,sperm morphology,DNA damage,and cytoplasmic remodeling were illustrated in this study to provide fresh perspectives on the mechanisms of male infertility.A total of 205 infertile males were allocated into 5 groups according to the percentage of spermatozoa exhibiting abnormal morphology within their samples.Sperm concentration,motility,abnormal sperm morphology,cytoplasmic droplets(CDs),and excess residual cytoplasm(ERC)were analyzed according to the World Health Organization manual(2010).Sperm nuclear vacuoles(NVs)were determined by propidium iodide(PI)staining.Sperm protamine expressions(P1 and P2)were detected by western blot.DNA damage was measured by acridine orange test(AOT)to calculate the proportion of sperm with single-strand DNA breaks(SSBs).Our data showed that sperm concentration and motility in infertile males significantly decreased with the severity of abnormal sperm morphology(both P<0.01).P1 level,P1/P2 ratio,and SSB rate increased with the severity of sperm dysmorphology,whilst the P2 level decreased(all P<O.01).NVs,CDs,and ERC were more common in males with sperm dysmorphology and positively correlated with the SSB rate(all P<O.01).The relationships between the SSB rate and the P1/P2 ratio were also significant(P<0.01).Aberrant protamination may cause sperm dysmorphology and compromise male fertility by impairing sperm's nucleus and cytoplasm maturation,with the P1/P2 ratio potentially serving as a valuable indicator of sperm quality and male fertility.
基金supported by the Institut Polaire Fran?ais,IPEV project 330 to OCPB was funded by an Arctic Field Grant(No.256934)from the Research Council of Norway+1 种基金by a Ph.D grant from University of La RochelleFH was financially supported by a grant from the Swiss National Science Foundation(No.PP00P3_139011)
文摘Background:Collecting and studying live sperm is central to many important fields of biology.Yet,a simple method to collect live sperm is lacking in wild seabird species.Here,we describe a non?invasive method to collect viable sperm samples based on a simple massage technique applied to male Black?legged Kittiwakes(Rissa tridactyla).Methods:We studied a colony breeding at Kongsfjorden,Svalbard and successfully obtained sperm samples from 32 males.With a subset of samples(n = 12 males),we compared the suitability of several extenders(0.9% NaCl,PBS,Earle's balance salt solution,Dulbecco's modified Eagle medium) in maintaining sperm alive long enough for analyses.With another 18 ejaculates,we conducted computer assisted sperm analyses using the CASA plugin for ImageJ.We provide details about the settings to be used for such analyses.Lastly,droplets from 20 ejaculates were smeared on glass slides and preserved with formalin to characterize sperm morphology in terms of total sperm length,sperm head length,midpiece length and flagellum length,and percentage of abnormal sperm.Results:With this method and under field conditions,we were able to obtain sufficient amounts of live sperm to assess traits related to sperm quality(e.g.sperm morphology,percentage of motile sperm,sperm velocity).We found that two extenders,Earle's balanced salt solution and Dulbecco modified Eagle's medium,yielded similarly good results.Additionally,we investigated whether specific behaviours were associated with successful sperm collection and whether sperm collection success depended on how long before laying sperm collection was attempted.Finally,we provide mean values for sperm morphology,sperm swimming ability and percentage of motile sperm,which may prove useful for future comparative analyses,and we report high levels of sperm abnormality and within?ejaculate variation in sperm morphology.Conclusions:We discuss the high percentage of abnormal sperm and high within?ejaculate variation in sperm morphology in light of sperm competition theory and conclude that these figures are likely due to relaxed post?cop?ulatory sexual selection,kittiwakes being strictly monogamous.Finally,we suggest that this method could be applied to other seabird species sharing similar ecology.
文摘The study investigated the intervention of caffeine on the effect of Nigerian Bonnylight crude oil (NBLCO) on sperm motility and morphology of diabetic Wistar rats. Eighty adult male rats (180 - 200 g body weight) were randomly divided into eight groups of 10 animals each. The control group received 3 mL/Kg body weight of distilled water, ND Caf. group received 20 mL/kg body weight of caffeine, NDCO group received 3 mL/Kg body weight of NBLCO, ND Caf. + CO group received 20 and 3 mL/Kg body weight of caffeine and NBLCO respectively, diabetic group received 3 mL/Kg body weight of distilled water, D Caf. group received 20 mL/Kg body weight of caffeine, D CO received 3 mL/Kg body weight of NBLCO and D Caf. + CO group received 20 and 3 mL/Kg body weight of caffeine and NBLCO respectively, by oral gavaging for 28 days. The results showed that independent administration of caffeine and NBLCO to both non-diabetic and diabetic rats significantly (p < 0.05) altered sperm concentration, morphology and motility. Administration of NBLCO aggravated the worsening condition by significantly (p < 0.05) reducing sperm concentration, motility and increasing abnormal sperms in diabetic rats. Although caffeine significantly (p < 0.05) reduced motility in non-diabetic rats, it did cause any significant alteration in diabetic rats. Diabetes mellitus and NBLCO significantly (p < 0.05) reduced all indices that promote sperm motility while significantly (p < 0.05) increasing indices that do not promote motility. It can be concluded that diabetes mellitus and NBLCO administration have shown the tendency to promote male infertility by reducing sperm motility and adversely altering the morphology of sperm in male Wistar rats, which was ameliorated by caffeine intervention.
文摘In this study, we compared conventional sperm selection with high-magnification morphology based on the motile sperm organellar morphology examination （MSOME） criteria, and hyaluronic acid （HA） binding for sperm chromosome aneuploidy and DNA fragmentation rates. Semen from 50 severe male factor cases was processed through density gradient centrifugation, and subjected to sperm selection by using the conventional method （control）, high magnification at x6650 or HA binding. Aneuploidy was detected by fluorescence in situ hybridization with probes for chromosomes 13, 18, 21, X and Y, and DNA fragmentation by the terminal deoxynucleotidyl transferase dUTP nick end labelling （TUNEL） method. Spermatozoa selected under high-magnification had a lower DNA fragmentation rate （2.6% vs. 1.7%; P=0.032）, with no significant difference in aneuploidy rate （0.8% vs0.7%; P=0.583）, than those selected by the HA binding method. Spermatozoa selected by both methods had much lower aneuploidy and DNA fragmentation rate than the controls （7% aneuploidy and 26.8% DNA fragmentation rates, respectively）. In the high-magnification group, the aneuploidy rate was lower when the best spermatozoa were selected than when only the second-best spermatozoa were available for selection, but the DNA fragmentation rate was not different. In conclusion, sperm selection under high magnification was more effective than under HA binding in selecting spermatozoa with low DNA fragmentation rate, but the small difference （0.9%） might not be clinically meaningful. Both methods were better than the conventional method of sperm selection.
基金supported by the National Natural Science Foundation of China(No.32100689)the National Key Research and Development Program of China(No.2021YFC2700202,No.2022YFA0806303,and No.2022YFC2702601)+1 种基金the Global Select Project(No.DJK-LX-2022010)of the Institute of Health and MedicineHefei Comprehensive National Science Center,and the Joint Fund for New Medicine of USTC(No.YD9100002034).
文摘Multiple morphological abnormalities of sperm flagella(MMAF)is a severe form of asthenoteratozoospermia,characterized by morphological abnormalities and reduced motility of sperm,causing male infertility.Although approximately 60%of MMAF cases can be explained genetically,the etiology of the remaining cases is unclear.Here,we identified two novel compound heterozygous variants in the gene,dynein axonemal heavy chain 10(DNAH10),in three patients from two unrelated Pakistani families using whole-exome sequencing(WES),including one compound heterozygous mutation(DNAH10:c.9409C>A[p.P3137T];c.12946G>C[p.D4316H])in family 1 and another compound heterozygous mutation(DNAH10:c.8849G>A[p.G2950D];c.11509C>T[p.R3687W])in family 2.All the identified variants are absent or rare in public genome databases and are predicted to have deleterious effects according to multiple bioinformatic tools.Sanger sequencing revealed that these variants follow an autosomal recessive mode of inheritance.Hematoxylin and eosin(H&E)staining revealed MMAF,including sperm head abnormalities,in the patients.In addition,immunofluorescence staining revealed loss of DNAH10 protein signals along sperm flagella.These findings broaden the spectrum of DNAH10 variants and expand understanding of the genetic basis of male infertility associated with the MMAF phenotype.
文摘Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both developing and developed countries. Other optional tests or sophisticated technologies have not been widely applied. The current review addresses important changes in the analysis of semen as described in the new World Health Organization （WHO） manual for semen analysis. The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality. Apart from the above mentioned changes, the initial evaluation and handling methods remain, in most instances, the same as in previous editions. Furthermore, the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology. WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995. The external qualitY control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training. This review reports on current sperm functional tests, such as the induced acrosome reaction, and sperm-zona pellucida binding assays, as well as the impact of sperm quality in terms of DNA integrity, and the relationship of sperm function tests to sperm morphology.
文摘Objective:To study the effect of mobile emitted radio frequency electromagnetic radiation(RF-EMR)on sperm parameters(sperm count,viability,motility,progressivity,and morphology)and ameliorative effect of pomegranate juice in rats.Methods:Thirty male Sprague Dawley rats were divided into five groups(n=6 in each group).GroupⅠwas neither exposed to RF-EMR nor given pomegranate juice,groupⅡ,ⅢandⅣwere exposed to mobile emitted RF-EMR for 60 min/day for 90 days.After 90-day exposure to RF-EMR,groupⅢwas supplemented with pomegranate juice for 90 days(1 mL/day)and groupⅣwas allowed to recover for 90 days without supplementation of pomegranate juice.GroupⅤwas supplemented with pomegranate juice for 90 days without exposure to RF-EMR.At the end of intervention,epididymal sperm parameters(sperm count,viability,motility,progressivity,and morphology)were measured.Results:The microscopic examination of sperm parameters such as sperm count,sperm viability,sperm motility,progressivity were significantly decreased in groupⅡ(the mobile RF-EMR group)compared to groupⅠ(the control group)(P<0.05).In addition,sperm morphology was also significantly altered(abnormal)in groupⅡcompared to groupⅠ(P<0.05).However,the sperm parameters including sperm morphology were significantly altered in groupⅢ(mobile RF-EMR+pomegranate juice)compared to groupⅡ(P<0.05).The sperm parameters including sperm morphology were not significantly altered in groupⅣ(the mobile RF-EMR recovery group)compared to groupⅡ(P>0.05).The sperm parameters were non-significantly increased in group V(the pomegranate juice group)compared to groupⅠ(P>0.05).Conclusions:Mobile RF-EMR exposure reduces the sperm parameters,which,however,can be reversed by treatment with pomegranate juice,indicating that pomegranate juice can be used as a nutritional supplement to improve sperm quality.
文摘Objective:To determine the relationship between teratozoospermia and sperm DNA fragmentation(SDF)in the human ejaculate.Methods:This retrospective study included 100 normozoospermic men as a control cohort(abnormal forms>14%),210 patients with a high level of abnormal forms(≤4%)and 65 patients presenting with a moderate level of abnormal forms(>4%to≤14%)based on the World Health Organization definitions.Sperm morphology was assessed using bright field microscopy.Sperm DNA fragmentation was assessed using the sperm chromatin dispersion assay.Non-parametric analyses were conducted to examine the relationship between abnormal sperm morphology and sperm DNA fragmentation;receiver operating characteristic(ROC)analyses were conducted to assess sensitivity and specificity of this relationship.Results:A correlation analysis revealed that the higher the proportion of abnormal spermatozoa in the ejaculate,the higher the level of SDF(Spearman's Rho=-0.230;P<0.001).Significant differences in the proportion of SDF were found when all cohorts were compared(P<0.001);these significant differences were also retained when the different cohorts were compared pairwise.ROC analysis showed a moderate but significant predictive value for SDF to differentiate patients with different levels of teratozoospemia.Conclusions:Although analysis of a more continuous range of values for teratozoospermia would help further clarify any causal relationship with SDF,there is clearly a synergistic or coincident affiliation between these variables that needs to be acknowledged by the clinician when interpreting the spermiogram.
文摘Aim： To investigate the reversibility of the effect of nandrolone decanoate （ND） on sperm parameters after discontinuing the drug. Methods： Three groups of rats received peanut oil （control）, low and high doses of ND for 14 weeks, respectively. Each group was divided into subgroups A and B, in which rats were killed 14 and 28 weeks after initiating the injection, respectively. Results： Sperm count and motile sperm fraction were decreased in the subgroups A and B that received low and high doses of ND in comparison with the controls A and B, respectively. The sperm count and motile sperm fraction increased in the subgroups B that received low and high doses of ND in comparison with their corresponding subgroups A. The number of normal morphology sperm was decreased significantly in subgroups A receiving low and high doses of ND in comparison with the control subgroup A. However, this parameter was not decreased in subgroups B receiving low and high doses in comparison with the control subgroup B. The normal sperm morphology did not show any significant differences in the subgroups B in comparison with their corresponding subgroups A. Conclusion： The 14-week injection of low and high doses of ND decreases sperm quality and quantity in rats. These parameters were improved after discontinuing ND, but not recovered completely even when they are left untreated for 14 weeks.
文摘Objective To investigate the age-long claim by the locales that the food thickener, M. urens seed, has antispermatogenic, hence, antifertility effects in man. Methods Eight-week old male Albino rats were used as the mammalian model for this study. They were assigned to four groups of 6 rats each and treatment with the ethanol extract was for a period of 14 d. The treatment regimes were 70 mg/kg, 140 mg/kg, 210 mg/kg and 0 mg/kg BW in groups A, B, C and D, respectively. Extracts were prepared by Soxhlet extraction using 80% ethanol as the extracting solvent. The stock solution was prepared by dissolving 1 g of the paste extract in 10 ml corn oil （vehicle） to make up 100 mg/ml concentration. At the end of the treatment, sperm from the distal caudal epididymis was collected and analyzed for sperm count, sperm motility and sperm morphology. Results Significant reduction was observed in sperm count and sperm motility （P〈0.05）. The mean sperm count for group A was 6.27±0.02×10^6, for group B was 6.16±0.02×10^6 and group C had 6.0±0.0×10^6 sperm cells The control （group D） had a mean sperm count of 6.50±0.09×10^6 which was higher than that of any treated group. Results of the sperm motility test gave the following mean rates for motile sperm cells after treatment： group A, 57.6±% 2.1; group B, 50.0±4.0; group C, 45.0±4.0. The control had the highest mean motility rate of 72.3±2.1. The observed sperm abnormalities included unusual head with large acrosome, looped tailpiece, mid piece with distal droplet, pin head, pyriform head and long hook.Conclusion The anti-spermatogenic effects of the extract on the sperm in the Albino rat may lead to reduction of fertility.
基金supported by Guangdong Science and Technology Department (No.2003B30506No.C31302)
文摘Objective To evaluate the effect of the accessory gential gland on the male fertility. Methods The relevant seminal parameters of 32 normal donors and 35 patients with olioasthenospermia were measured by seminal routine analysis, sperm morphology and accessory, gentital glands markers.Results The percentage of normal spermatozoa, the function of sperm penetration through cervical mucus and the enzyme activity of neutral α-1,4-glucosidase significantly declined as compared with the control. There was a significant negative relationship between the semen volume and the sperm motility for the oligoasthenospermia （r= -0.415, P〈0.05）, and a significant positive relationship between the sperm count and the percentage of normal sperm （r=0.393, P〈0.05）.Conclusion The abnormal function of epididymis exists in patients with oligoasthenospermia. Sperm function,for the oligoasthenospermia declines with an increase of the percentage of the abnormal sperm. The poorer the spermatogenises function of the testis is, the higher the percentage of the abnormal sperm appears.
文摘Objective To introduce a rapid simple staining method for sperm morphology.Methods Liquifiedsemen was on the glass, fixed by methanol, treated with phosphonic acid buffer, stained by A, B staining solution separately, and rinsed by running water. Results After staining, the whole sper-matozoid was very clear. The cephalic zone of spermatozoid head , acrosome area , was colored pink; its caudal zone colored violet red or violet blue. The body and tail of spermatozoid colored pink or light blue. Normal and abnormal morphology of spermatozoid could be differentiated easily. The sloughed spermatogenic cells and white blood cells could be differentiated also. Conclusion This rapid staining method of semen smear can produce the same effect of other usual staining method , but the staining time is shorter and the procedure is simpler.
基金This study was supported by grants from the National Natural Science Foundation of China(81771645 and 81471432 to YQT),and Graduate Research and Innovation Projects of Central South University(Grant 2017zzts071 to CFT).
文摘The syndrome of multiple morphological abnormalities of the sperm flagella(MMAF)is a specific kind of asthenoteratozoospermia with a mosaic of flagellar morphological abnormalities(absent,short,bent,coiled,and irregular flagella).MMAF was proposed in 2014 and has attracted increasing attention;however,it has not been clearly understood.In this review,we elucidate the definition of MMAF from a systematical view,the difference between MMAF and other conditions with asthenoteratozoospermia or asthenozoospermia(such as primary mitochondrial sheath defects and primary ciliary dyskinesia),the knowledge regarding its etiological mechanism and related genetic findings,and the clinical significance of MMAF for intracytoplasmic sperm injection and genetic coun sell ng.This review provides the basic kno wledge for MMAF and puts forward some suggestions for further investigations.
基金supported by grant from the National Key Research and Development Program of China(No.2017YFC1002003).
文摘Numerous genes have been associated with multiple morphological abnormalities of the sperm flagella(MMAF),which cause severe asthenozoospermia and lead to male infertility,while the causes of approximately 50%of MMAF cases remain unclear.To reveal the genetic causes of MMAF in an infertile patient,whole-exome sequencing was performed to screen for pathogenic genes,and electron microscope was used to reveal the sperm flagellar ultrastructure.A novel heterozygous missense mutation in the outer dense fiber protein 2(ODF2)gene was detected,which was inherited from the patient’s mother and predicted to be potentially damaging.Transmission electron microscopy revealed that the outer dense fibers were defective in the patient’s sperm tail,which was similar to that of the reported heterozygous Odf2 mutation mouse.Immunostaining of ODF2 showed severe ODF2 expression defects in the patient’s sperm.Therefore,it was concluded that the heterozygous mutation in ODF2 caused MMAF in this case.To evaluate the possibility of assisted reproductive technology(ART)treatment for this patient,intracytoplasmic sperm injection(ICSI)was performed,with the help of a hypo-osmotic swelling test and laser-assisted immotile sperm selection(LAISS)for available sperm screening,and artificial oocyte activation with ionomycin was applied to improve the fertilization rate.Four ICSI cycles were performed,and live birth was achieved in the LAISS-applied cycle,suggesting that LAISS would be valuable in ART treatment for MMAF.
基金This work was supported by the National Natural Science Foundation of China(No.32070850)the National Natural Science Foundation of China(No.31630050,31890780,and 32061143006)+2 种基金the National Key Research and Developmental Program of China(2018YFC1003900,2019YFA0802600,and 2016YFC1000600)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDB19000000)the Fundamental Research Funds for the Central Universities(No.YD2070002006).
文摘Multiple morphological abnormalities of the sperm flagella(MMAF)is a specific type of asthenoteratozoospermia,presenting with multiple morphological anomalies in spermatozoa,such as absent,bent,coiled,short,or irregular caliber flagella.Previous genetic studies revealed pathogenic mutations in genes encoding cilia and flagella-associated proteins(CFAPs;e.g.,CFAP43,CFAP44,CFAP65,CFAP69,CFAP70,and CFAP251)responsible for the MMAF phenotype in infertile men from different ethnic groups.However,none of them have been identified in infertile Pakistani males with MMAF.In the current study,two Pakistani families with MMAF patients were recruited.Whole-exome sequencing(WES)of patients and their parents was performed.WES analysis reflected novel biallelic loss-of-function mutations in CFAP43 in both families(Family 1:ENST00000357060.3,p.Arg300Lysfs*22 and p.Thr526Serfs*43 in a compound heterozygous state;Family 2:ENST00000357060.3,p.Thr526Serfs*43 in a homozygous state).Sanger sequencing further confirmed that these mutations were segregated recessively in the families with the MMAF phenotype.Semiquantitative reverse-transcriptase polymerase chain reaction(qRT-PCR)was carried out to detect the effect of the mutation on mRNA of the affected gene.Previous research demonstrated that biallelic loss-of-function mutations in CFAP43 accounted for the majority of all CFAP43-mutant MMAF patients.To the best of our knowledge,this is the first study to report CFAP43 biallelic loss-of-function mutations in a Pakistani population with the MMAF phenotype.This study will help researchers and clinicians to understand the genetic etiology of MMAF better.
