CatSper is a unique Ca2＋ channel-like protein family exclusively expressed in the testis and sperm, and plays important roles in sperm motility, capacitation, acrosome reaction and sperm-egg interactions. Here we stu...CatSper is a unique Ca2＋ channel-like protein family exclusively expressed in the testis and sperm, and plays important roles in sperm motility, capacitation, acrosome reaction and sperm-egg interactions. Here we studied the mechanism of regulation of CatSper2-dependent Ca〉 influx, extracellular and intracellular Ca2＋on sperm hyperactivated motility. The siRNA duplexes were transfected into the sperm cells by electroporation （EP） to silence the expression of CatSper2. The results for targeted disruption of CatSper2 showed the highest silence efficiency 77.7% （P〈0.05）, the hyperactivated sperm rate calculated by computer-assisted semen analysis （CASA） analysis of interferenced sperm was significantly lower 11.1% than the control 99.2%. Flow cytometry （FCM） detection of the intracellular Ca2＋ concentration of interferenced sperm was 50 nmol L-t higher than the normal. It was remarkably lower than hyperactivated sperm with 200-1 000 nmol L-1 （P〈0.05）. It was not sufficient to evoke hyperactivation. To trigger hyperactivation, the sperm were incubated in 50 pmol L-t thimerosal or 5 mmol L-1 procaine, it sharply increased the intracellular Ca2＋ via two different channels. CASA and FCM detection indicated that intracellular Ca〉 is required for initiating hyperactivation while extracellular Ca2＋ is essential to maintain the process. We concluded that to mediate sperm hyperactivation, it is essential to inhibit Ca〉peak present with targeted disruption of CatSper2 expression. This provides important prospective for further exploration of signal channel of sperm hyperactivated motility, potential factors for male infertility and provide further reference to the exploration of male contraceptive drug targets of male reproduction.展开更多
Introduction: Men without sperm (azoospermic) make up about 15% of all infertile men in reproductive age of fatherhood. Male infertility is a health and social problem in many communities. Subjects and Methods: This r...Introduction: Men without sperm (azoospermic) make up about 15% of all infertile men in reproductive age of fatherhood. Male infertility is a health and social problem in many communities. Subjects and Methods: This retrospective study was carried from 2010 to 2015. Initially, 907 men were included in the study among whom 109 (12.02%), 346 (38.15%) and 452 (49.83%) were azoospermic, oligospermic and normospermic, respectively. This study only investigated the association between age, BMI, semen volume, liquefaction time and pus cells among normospermic and azoospermic men. Standard semen analysis was performed and subjects were categorized by age into Results: The 562 subjects of the study had means (±sd) age of 42.6 (±7.10) years and BMI of 27.0 (4.1) kg/m2 respectively. In all, 109 (19.4%) were azoospermic. Overweight and obese men were, respectively, more than 1? and about 2? times more likely to be azoospermic compared to normal weight men. Azoospermic men were significantly heavier than men with normal sperm count (t = -0.34;P-value = 0.003). Among those with normal weight, liquefaction time was significantly shorter (t = 5.49, P-value = 0.000001) in azoospermic (28.70 min.) than in men with normal sperm count (31.82 min.). Obese azoospermic men were about 4 times as likely to have high pus cells in semen than normal weight azoospermic men (OR = 3.82;95% CI: 0.39, 37.01). Multivariate regression analysis shows a strong but negative coefficient correlation between sperm concentration and BMI (coef. = -0.48, Std Err. = 0.25, P-value = 0.05, 95% CI: -0.96, 0.10). Conclusion: Our findings suggest that high BMI is associated with azoospermia. Azoospermia also did not seem to negatively impact liquefaction time.展开更多
Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by ...Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by assessing the post-selection cell quality.Semen from six Mazama deer was collected by electroejaculation after chemical restraint.The semen was analyzed in four samples:Fresh,DGC,SEMgood-non-apoptotic fraction,and SEMpoor-apoptotic fraction.The material was analyzed for motility and vigor(light microscopy),concentration(Neubauer chamber),semen morphology(phase contrast),and supravital staining test(eosin/nigrosine).The DGC method used 20 x 106 cells in 90%and 45%percoll®gradient.The MASS used 10 x 106 cells with 20μl of iron nanoparticles attached to Annexin V and filtration in a magnetic separation column.Both processing methods(DGC and MASS)were effective in producing high-quality sperm samples,with a marked reduction in abnormalities from 41.83±10.25(fresh)to 14.83±3.17(DGC)and 12±3.01(SEMgood),with 80.3%±2.06 livings cells.These findings suggest that this nanotechnological method,using nanoparticles,effectively produces high-quality semen samples in cervids for use in assisted reproduction.展开更多
Background:Hormonal treatment and response as a predictor of sperm retrieval prior to microdissection testicular sperm extraction(micro-TESE)are not well established in the current literature.This study aimed to inves...Background:Hormonal treatment and response as a predictor of sperm retrieval prior to microdissection testicular sperm extraction(micro-TESE)are not well established in the current literature.This study aimed to investigate the hormonal response as a predictor of sperm retrieval among men with nonobstructive azoospermia(NOA).Methods:Seventy-seven consecutive patients who had testosterone levels≤14 nmol/L were treated medically with an aromatase inhibitor or recombinant human chorionic gonadotropin(rec-hCG)prior to micro-TESE and were included.Thirty-four(44.2%)had unexplained NOA(UNEX),25(32.5%)had Klinefelter syndrome(KS),8(10.4%)had a history of cryptorchidism(UDT),4(5.2%)had microdeletion of the Azoospermia factor C(AZFc),and 6(7.8%)were treated previously with chemotherapy.Baseline and post-treatment serum hormonal levels were documented.Pre-op testosterone levels were entered into binary logistic regressions with age,Follicle-stimulating hormone(FSH),and Luteinizing hormone(LH)levels to test for significance with sperm retrieval.We then built logistic regression models to identify predictors of successful surgical sperm retrieval(SSR).Results:Forty-five patients(58%)had successful retrieval.In 32 patients(42%),no sperm was retrieved.Both the mean pre-op testosterone and the mean testosterone change between the two groups were significant(p=0.02 and p=0.011,respectively).Receiver operating characteristic(ROC)analysis demonstrated an area under the curve(AUC)of 0.785(95%CI=0.685-0.886,p<0.001).The Youden index coefficient was calculated for KS and UNEX.The cut-off point for KS was established at 0.764(sensitivity=0.875,false positive rate[FPR]=0.111),and 0.215 for UNEX(sensitivity=0.438,FPR=0.222).We also observed a correlation between age and SSR(p=0.05).In KS patients,SSR was determined by pre-op testosterone levels irrespective of age.Conclusion:Pre-operative hormonal response is a predictor for SSR in NOA patients who were treated medically.This data may help during pre-operative counselling.展开更多
Optical tweezers technology has the characteristics of noncontact manipulation in three dimensions and steerable separation in solutions,and could be applied to obtain a separated sperm with high quality for intracyto...Optical tweezers technology has the characteristics of noncontact manipulation in three dimensions and steerable separation in solutions,and could be applied to obtain a separated sperm with high quality for intracytoplasmic sperm injection(ICSI).However,the effects of optical tweezers on sperm motility are still unclear.To elucidate the effects on sperm motility for optical tweezers,we systematically investigated the correlation between motility parameters and the parameters of optical tweezers(wavelength,power,trapping duration,and trapping orientation).Under three systems of optical tweezers with different laser wavelengths(1064,850,and 785 nm),the nine motility parameters of free swimming were mainly affected by trapping orientation(vertical/horizontal)and trapping duration.When 850 nm laser and 1064 nm laser are used,vertical trapping significantly reduces sperm free-swimming capability with prolonged exposure time,whereas horizontal trapping exhibits relatively minor interference on sperm motility.Notably,the 785 nm laser does not induce statistically significant changes in key parameters of sperm motility under any experimental conditions(trapping orientation and duration).For the rolling frequency of trapped sperm,horizontal trapping for three wavelengths has a negligible effect compared with vertical trapping,especially for the 785 nm laser.In conclusion,horizontal trapping can preserve sperm motility under low power(below 140mW at 1064 nm,below 100mW at 850nm,below 60mW at 785 nm)and short duration(below 4 min).This trapping duration is suffcient for the separation procedure of single live sperm in ICSI.This study provides critical parameter optimization guidelines for the safe application of optical tweezers technology in reproductive medicine.展开更多
Rationale:This case report describes a couple with recurrent fertilization failure despite undergoing multiple cycles of intracytoplasmic sperm injection(ICSI).The principal clinical concern was suspected oocyte activ...Rationale:This case report describes a couple with recurrent fertilization failure despite undergoing multiple cycles of intracytoplasmic sperm injection(ICSI).The principal clinical concern was suspected oocyte activation deficiency(OAD),in which fertilization is impeded due to the oocyte’s inability to initiate embryogenesis,commonly attributed to inadequate intracellular calcium(Ca^(2+))release following sperm injection.Patient concerns:The couple repeatedly experienced complete or near-complete fertilization failure in previous ICSI cycles,raising suspicion of an underlying oocyte activation defect.Diagnosis:Based on the repeated absence of fertilization post-ICSI and clinical history,a diagnosis of suspected OAD leading to recurrent ICSI fertilization failure was considered.Interventions:Artificial oocyte activation(AOA)using the calcium ionophore A23187 was performed.After ICSI,unfertilized oocytes were exposed to the ionophore to induce Ca^(2+)influx,simulating physiological calcium oscillations essential for oocyte activation.The efficacy of intervention was evaluated through subsequent embryonic development,morphological grading,and chromosomal integrity.Outcomes:Following AOA treatment,successful oocyte activation occurred,resulting in the formation of high-grade embryos with normal developmental progression.Chromosomal analysis revealed no detectable abnormalities,indicating genomic stability.Lessons:Calcium ionophore–mediated AOA may serve as an effective adjunct in cases of recurrent ICSI failure attributed to OAD.This case highlights the importance of individualized therapeutic strategies in assisted reproduction;however,further research is needed to refine protocols,validate broader clinical efficacy,and assess long-term safety,including potential epigenetic risks.展开更多
The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoingin vitro fertilization (IVF)has been a debated hot topic among reproductive specialists. Each approach presents distinc...The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoingin vitro fertilization (IVF)has been a debated hot topic among reproductive specialists. Each approach presents distinct advantages and disadvantages,with fresh sperm typically showing superior sperm quality, while frozen sperm offers logistical flexibility and a reliable backup forrepeated cycles. This review summarizes the latest advancements in sperm retrieval and cryopreservation techniques, providingpractitioners with a comprehensive analysis of each option’s strengths and limitations. Comparative studies indicate that, althoughfresh sperm often has better quality metrics, cryopreservation methods such as vitrification have significantly improved postthawoutcomes, making frozen sperm a viable choice in assisted reproductive technologies (ART). The findings show comparablerates for fertilization, implantation, clinical pregnancy, and live birth between fresh and frozen microdissection testicular spermextraction (micro-TESE) sperm in many cases, although patient-specific factors such as timing, cost-effectiveness, and proceduralconvenience should guide the final decision. Ultimately, the choice of using fresh or frozen sperm should align with the individualneeds and conditions of patients. This tailored approach, supported by the latest advancements, can optimize ART outcomes andprovide personalized reproductive care.展开更多
Azoospermia is characterized by the absence of sperm in the ejaculate and is categorized into obstructive azoospermia(OA)and nonobstructive azoospermia(NOA).For men with NOA,testicular sperm extraction(TESE)is the onl...Azoospermia is characterized by the absence of sperm in the ejaculate and is categorized into obstructive azoospermia(OA)and nonobstructive azoospermia(NOA).For men with NOA,testicular sperm extraction(TESE)is the only method to obtain sperm for assisted reproductive technology(ART).Given the rarity of these sperm and the unpredictable success of subsequent retrieval attempts,cryopreservation of microdissection-TESE-obtained sperm is essential.Effective cryopreservation prevents the need for repeated surgical procedures and supports future ART attempts.After first delving into the physiological and molecular aspects of sperm cryopreservation,this review aims to examine the current methods and devices for preserving small numbers of sperm.It presents conventional freezing and vitrification techniques,evaluating their respective strengths and limitations in effectively preserving rare sperm,and compares the efficacy of using fresh versus cryopreserved testicular sperm.展开更多
Primary ciliary dyskinesia(PCD)is a clinically rare,genetically and phenotypically heterogeneous condition characterized by chronic respiratory tract infections,male infertility,tympanitis,and laterality abnormalities...Primary ciliary dyskinesia(PCD)is a clinically rare,genetically and phenotypically heterogeneous condition characterized by chronic respiratory tract infections,male infertility,tympanitis,and laterality abnormalities.PCD is typically resulted from variants in genes encoding assembly or structural proteins that are indispensable for the movement of motile cilia.Here,we identified a novel nonsense mutation,c.466G>T,in cilia-and flagella-associated protein 300(CFAP300)resulting in a stop codon(p.Glu156*)through whole-exome sequencing(WES).The proband had a PCD phenotype with laterality defects and immotile sperm flagella displaying a combined loss of the inner dynein arm(IDA)and outer dynein arm(ODA).Bioinformatic programs predicted that the mutation is deleterious.Successful pregnancy was achieved through intracytoplasmic sperm injection(ICSI).Our results expand the spectrum of CFAP300 variants in PCD and provide reproductive guidance for infertile couples suffering from PCD caused by them.展开更多
Background Freezing-induced sperm damage,often associated with oxidative stress,can result in regulated cell death.Given that oxidative stress can trigger various forms of regulated cell death,the prevailing form duri...Background Freezing-induced sperm damage,often associated with oxidative stress,can result in regulated cell death.Given that oxidative stress can trigger various forms of regulated cell death,the prevailing form during sperm cryopreservation remains unknown.Our study aimed to investigate this issue using cashmere goats as a model.Results We found a significant increase in lyso-phospholipids in frozen-thawed sperm suggested ferroptosis.Assessment of cryopreserved sperm,with or without prior treatment with ferroptosis or apoptosis inhibitors,demonstrated the significant efficacy of ferroptosis inhibitors in reducing freezing damage.This implicates ferroptosis as the primary form of regulated cell death induced during sperm cryopreservation.Additionally,the positive rate of transferrin receptor protein 1 was significantly lower in fresh live sperm(47.8%)than in thawed live sperm(71.5%),and the latter rate was lower than that in dead sperm(82.5%).By contrast,cleaved caspase-3 positivity showed no significant difference between fresh live sperm and thawed live sperm but was notably lower in thawed live sperm than in dead sperm.Conclusions Our findings establish ferroptosis as the dominant regulated cell death form during goat sperm cryopreservation,providing novel insights into freezing-induced sperm damage mechanisms.These findings have significant implications for optimizing cryopreservation protocols and enhancing sperm viability after freezing-thawing.展开更多
Total or severe teratospermia affects the prognosis of fertility and causes serious problems for patients undergoing assisted reproduction[1].The pathophysiological mechanism of teratospermia is unclear.It has been sh...Total or severe teratospermia affects the prognosis of fertility and causes serious problems for patients undergoing assisted reproduction[1].The pathophysiological mechanism of teratospermia is unclear.It has been shown that patients with sperm parameters abnormalities and abnormal morphology have a high rate of fragmentation and sperm DNA decondensation[2,3],and that sperm DNA fragmentation analysis could be used as a predictor factor of fertility potential[4].展开更多
The analysis of the ejaculate,better known as spermiogram,represents the first and main step to identify whether a series of sperm quality parameters are within the norm and therefore are consistent with normal sperm ...The analysis of the ejaculate,better known as spermiogram,represents the first and main step to identify whether a series of sperm quality parameters are within the norm and therefore are consistent with normal sperm fertilizing capacity.Among these,sperm concentration and motility are the first parameters to be evaluated through an estimation carried out by expert examiners.展开更多
Depression currently affects about 280 million people worldwide and its prevalence has been increasing dramatically,especially among the young and people of reproductive age,which consequently leads to an increase in ...Depression currently affects about 280 million people worldwide and its prevalence has been increasing dramatically,especially among the young and people of reproductive age,which consequently leads to an increase in antidepressant consumption.Antidepressants are associated with sexual dysfunction in both men and women;however,their role in male fertility has been scarcely studied.Fluoxetine and sertraline,two serotonin reuptake inhibitors(SSRIs),are among the most prescribed antidepressants worldwide.To determine their possible effects,human sperm cells were exposed to either sertraline or fluoxetine at concentrations previously found in blood and seminal fluid of patients undergoing treatment.Spermatozoa were incubated for up to 24 h at 37℃ and 5%CO_(2),and important functional parameters such as sperm motility,viability,mitochondrial membrane potential,cellular reactive oxygen species(ROS)production,chromatin/DNA integrity,acrosome status,and tyrosine phosphorylation were assessed.At low levels,fluoxetine consistently decreased progressive motility throughout time while promoting fluctuations in ROS levels and sperm capacitation.Nevertheless,it did not affect viability,mitochondrial membrane potential,acrosome reaction nor chromatin/DNA integrity.Sertraline,on the other hand,had little to nonsignificant impact at low doses,but affected almost all tested parameters at supratherapeutic concentrations.Altogether,our results suggest that both antidepressants may impair sperm function,possibly through different mechanisms of action,but fluoxetine is the only exhibiting mild negative effects at doses found in vivo.展开更多
The advent of intracytoplasmic sperm injection,along with the realization that many men with azoospermia due to primary testicular failure may have a few spermatozoa in their testes,has resulted in the revolutionary p...The advent of intracytoplasmic sperm injection,along with the realization that many men with azoospermia due to primary testicular failure may have a few spermatozoa in their testes,has resulted in the revolutionary possibility of azoospermic men fathering their own genetic offspring.展开更多
High levels of sperm DNA fragmentation(SDF)are associated with reduced assisted reproductive technology(ART)outcomes.Currently,SDF is not included in routine clinical assessment of male partners of infertile couples,b...High levels of sperm DNA fragmentation(SDF)are associated with reduced assisted reproductive technology(ART)outcomes.Currently,SDF is not included in routine clinical assessment of male partners of infertile couples,but the 6th edition of the World Health Organization(WHO)manual for semen analysis included the SDF assessment in the chapter on extended semen examinations.展开更多
Reactive oxygen species(Ros)play a dual role in mammalian spermatozoa.At high levels,they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein,lipids,and sperm D...Reactive oxygen species(Ros)play a dual role in mammalian spermatozoa.At high levels,they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein,lipids,and sperm DNA.This oxidative damage is associated with male infertility.On the other hand,when RoS are produced at low levels,they participate in the redox signaling necessary for sperm capacitation.Capacitation-associated RoS are produced by the sperm oxidase,whose identity is still elusive,located in the plasma membrane of the spermatozoon.Ros,such as superoxide anion,hydrogen peroxide,nitric oxide,and peroxynitrite,activate protein kinases and inactivate protein phosphatases with the net increase of specific phosphorylation events.Peroxiredoxins(PRDXs),antioxidant enzymes that fight against oxidative stress,regulate redox signaling during capacitation.Among them,PRDX6,which possesses peroxidase and calcium-independent phospholipase A,(iPLA,)activities,is the primary regulator of redox signaling and the antioxidant response in human spermatozoa.The lysophosphatidic acid signaling is essential to maintain sperm viability by activating the phosphatidylinositol 3-kinase/protein kinase(PI3K/AKT)pathway,and it is regulated by PRDX6 iPLA2,protein kinase C(PKC),and receptor-type protein tyrosine kinase.The understanding of redox signaling is crucial to pave theway fornovel diagnostic tools and treatments of male infertility.展开更多
Kamil Gill,PhD,Kamil Gill is a biologist who graduated from the University of Szczecin(Poland)with a Master degree.Dr.Gill earned his scientific degree from the Pomeranian Medical University in Szczecin(PUM).His docto...Kamil Gill,PhD,Kamil Gill is a biologist who graduated from the University of Szczecin(Poland)with a Master degree.Dr.Gill earned his scientific degree from the Pomeranian Medical University in Szczecin(PUM).His doctoral thesis was entitled:"Molecular markers of male infertility-evaluation of selected human sperm chromatin parameters and its influence on the zygote and embryo development in assisted reproduction conditions".展开更多
Except in cases of hypogonadotropic hypogonadism,the use of medical therapy before microsurgical testicular sperm extraction(micro-TESE)is controversial.In some studies,hormone therapy has been shown to improve the po...Except in cases of hypogonadotropic hypogonadism,the use of medical therapy before microsurgical testicular sperm extraction(micro-TESE)is controversial.In some studies,hormone therapy has been shown to improve the possibility of sperm retrieval during micro-TESE and even lead to the presence of sperm in the ejaculate in some cases,thereby obviating the need for micro-TESE.However,their routine use before micro-TESE in cases of nonobstructive azoospermia(NOA)being associated with hypergonadotropic hypogonadism and eugonadism(normogonadotropic condition)has not been supported with robust evidence.In this review,we discuss different types of medical therapy used before micro-TESE for NOA,their risks and benefits,and the available evidence surrounding their use in this setting.展开更多
基金supported by the National Key Technology R&D Program of China (2006BAD04A12)
文摘CatSper is a unique Ca2＋ channel-like protein family exclusively expressed in the testis and sperm, and plays important roles in sperm motility, capacitation, acrosome reaction and sperm-egg interactions. Here we studied the mechanism of regulation of CatSper2-dependent Ca〉 influx, extracellular and intracellular Ca2＋on sperm hyperactivated motility. The siRNA duplexes were transfected into the sperm cells by electroporation （EP） to silence the expression of CatSper2. The results for targeted disruption of CatSper2 showed the highest silence efficiency 77.7% （P〈0.05）, the hyperactivated sperm rate calculated by computer-assisted semen analysis （CASA） analysis of interferenced sperm was significantly lower 11.1% than the control 99.2%. Flow cytometry （FCM） detection of the intracellular Ca2＋ concentration of interferenced sperm was 50 nmol L-t higher than the normal. It was remarkably lower than hyperactivated sperm with 200-1 000 nmol L-1 （P〈0.05）. It was not sufficient to evoke hyperactivation. To trigger hyperactivation, the sperm were incubated in 50 pmol L-t thimerosal or 5 mmol L-1 procaine, it sharply increased the intracellular Ca2＋ via two different channels. CASA and FCM detection indicated that intracellular Ca〉 is required for initiating hyperactivation while extracellular Ca2＋ is essential to maintain the process. We concluded that to mediate sperm hyperactivation, it is essential to inhibit Ca〉peak present with targeted disruption of CatSper2 expression. This provides important prospective for further exploration of signal channel of sperm hyperactivated motility, potential factors for male infertility and provide further reference to the exploration of male contraceptive drug targets of male reproduction.
文摘Introduction: Men without sperm (azoospermic) make up about 15% of all infertile men in reproductive age of fatherhood. Male infertility is a health and social problem in many communities. Subjects and Methods: This retrospective study was carried from 2010 to 2015. Initially, 907 men were included in the study among whom 109 (12.02%), 346 (38.15%) and 452 (49.83%) were azoospermic, oligospermic and normospermic, respectively. This study only investigated the association between age, BMI, semen volume, liquefaction time and pus cells among normospermic and azoospermic men. Standard semen analysis was performed and subjects were categorized by age into Results: The 562 subjects of the study had means (±sd) age of 42.6 (±7.10) years and BMI of 27.0 (4.1) kg/m2 respectively. In all, 109 (19.4%) were azoospermic. Overweight and obese men were, respectively, more than 1? and about 2? times more likely to be azoospermic compared to normal weight men. Azoospermic men were significantly heavier than men with normal sperm count (t = -0.34;P-value = 0.003). Among those with normal weight, liquefaction time was significantly shorter (t = 5.49, P-value = 0.000001) in azoospermic (28.70 min.) than in men with normal sperm count (31.82 min.). Obese azoospermic men were about 4 times as likely to have high pus cells in semen than normal weight azoospermic men (OR = 3.82;95% CI: 0.39, 37.01). Multivariate regression analysis shows a strong but negative coefficient correlation between sperm concentration and BMI (coef. = -0.48, Std Err. = 0.25, P-value = 0.05, 95% CI: -0.96, 0.10). Conclusion: Our findings suggest that high BMI is associated with azoospermia. Azoospermia also did not seem to negatively impact liquefaction time.
文摘Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by assessing the post-selection cell quality.Semen from six Mazama deer was collected by electroejaculation after chemical restraint.The semen was analyzed in four samples:Fresh,DGC,SEMgood-non-apoptotic fraction,and SEMpoor-apoptotic fraction.The material was analyzed for motility and vigor(light microscopy),concentration(Neubauer chamber),semen morphology(phase contrast),and supravital staining test(eosin/nigrosine).The DGC method used 20 x 106 cells in 90%and 45%percoll®gradient.The MASS used 10 x 106 cells with 20μl of iron nanoparticles attached to Annexin V and filtration in a magnetic separation column.Both processing methods(DGC and MASS)were effective in producing high-quality sperm samples,with a marked reduction in abnormalities from 41.83±10.25(fresh)to 14.83±3.17(DGC)and 12±3.01(SEMgood),with 80.3%±2.06 livings cells.These findings suggest that this nanotechnological method,using nanoparticles,effectively produces high-quality semen samples in cervids for use in assisted reproduction.
文摘Background:Hormonal treatment and response as a predictor of sperm retrieval prior to microdissection testicular sperm extraction(micro-TESE)are not well established in the current literature.This study aimed to investigate the hormonal response as a predictor of sperm retrieval among men with nonobstructive azoospermia(NOA).Methods:Seventy-seven consecutive patients who had testosterone levels≤14 nmol/L were treated medically with an aromatase inhibitor or recombinant human chorionic gonadotropin(rec-hCG)prior to micro-TESE and were included.Thirty-four(44.2%)had unexplained NOA(UNEX),25(32.5%)had Klinefelter syndrome(KS),8(10.4%)had a history of cryptorchidism(UDT),4(5.2%)had microdeletion of the Azoospermia factor C(AZFc),and 6(7.8%)were treated previously with chemotherapy.Baseline and post-treatment serum hormonal levels were documented.Pre-op testosterone levels were entered into binary logistic regressions with age,Follicle-stimulating hormone(FSH),and Luteinizing hormone(LH)levels to test for significance with sperm retrieval.We then built logistic regression models to identify predictors of successful surgical sperm retrieval(SSR).Results:Forty-five patients(58%)had successful retrieval.In 32 patients(42%),no sperm was retrieved.Both the mean pre-op testosterone and the mean testosterone change between the two groups were significant(p=0.02 and p=0.011,respectively).Receiver operating characteristic(ROC)analysis demonstrated an area under the curve(AUC)of 0.785(95%CI=0.685-0.886,p<0.001).The Youden index coefficient was calculated for KS and UNEX.The cut-off point for KS was established at 0.764(sensitivity=0.875,false positive rate[FPR]=0.111),and 0.215 for UNEX(sensitivity=0.438,FPR=0.222).We also observed a correlation between age and SSR(p=0.05).In KS patients,SSR was determined by pre-op testosterone levels irrespective of age.Conclusion:Pre-operative hormonal response is a predictor for SSR in NOA patients who were treated medically.This data may help during pre-operative counselling.
基金supported by the Natural Science Foundation of Anhui Province in China(2508085MF166)Research Fund of Anhui Institute of Translational Medicine(2024zh-03)+2 种基金Key Scientific Research Foundation of Education Department of Anhui Province(2023AH040083)National Natural Science Foundation of China(12404353)Cultivation Project of Training Young and Middle-aged Teachers in Universities of Anhui Province(DTR2023013).
文摘Optical tweezers technology has the characteristics of noncontact manipulation in three dimensions and steerable separation in solutions,and could be applied to obtain a separated sperm with high quality for intracytoplasmic sperm injection(ICSI).However,the effects of optical tweezers on sperm motility are still unclear.To elucidate the effects on sperm motility for optical tweezers,we systematically investigated the correlation between motility parameters and the parameters of optical tweezers(wavelength,power,trapping duration,and trapping orientation).Under three systems of optical tweezers with different laser wavelengths(1064,850,and 785 nm),the nine motility parameters of free swimming were mainly affected by trapping orientation(vertical/horizontal)and trapping duration.When 850 nm laser and 1064 nm laser are used,vertical trapping significantly reduces sperm free-swimming capability with prolonged exposure time,whereas horizontal trapping exhibits relatively minor interference on sperm motility.Notably,the 785 nm laser does not induce statistically significant changes in key parameters of sperm motility under any experimental conditions(trapping orientation and duration).For the rolling frequency of trapped sperm,horizontal trapping for three wavelengths has a negligible effect compared with vertical trapping,especially for the 785 nm laser.In conclusion,horizontal trapping can preserve sperm motility under low power(below 140mW at 1064 nm,below 100mW at 850nm,below 60mW at 785 nm)and short duration(below 4 min).This trapping duration is suffcient for the separation procedure of single live sperm in ICSI.This study provides critical parameter optimization guidelines for the safe application of optical tweezers technology in reproductive medicine.
文摘Rationale:This case report describes a couple with recurrent fertilization failure despite undergoing multiple cycles of intracytoplasmic sperm injection(ICSI).The principal clinical concern was suspected oocyte activation deficiency(OAD),in which fertilization is impeded due to the oocyte’s inability to initiate embryogenesis,commonly attributed to inadequate intracellular calcium(Ca^(2+))release following sperm injection.Patient concerns:The couple repeatedly experienced complete or near-complete fertilization failure in previous ICSI cycles,raising suspicion of an underlying oocyte activation defect.Diagnosis:Based on the repeated absence of fertilization post-ICSI and clinical history,a diagnosis of suspected OAD leading to recurrent ICSI fertilization failure was considered.Interventions:Artificial oocyte activation(AOA)using the calcium ionophore A23187 was performed.After ICSI,unfertilized oocytes were exposed to the ionophore to induce Ca^(2+)influx,simulating physiological calcium oscillations essential for oocyte activation.The efficacy of intervention was evaluated through subsequent embryonic development,morphological grading,and chromosomal integrity.Outcomes:Following AOA treatment,successful oocyte activation occurred,resulting in the formation of high-grade embryos with normal developmental progression.Chromosomal analysis revealed no detectable abnormalities,indicating genomic stability.Lessons:Calcium ionophore–mediated AOA may serve as an effective adjunct in cases of recurrent ICSI failure attributed to OAD.This case highlights the importance of individualized therapeutic strategies in assisted reproduction;however,further research is needed to refine protocols,validate broader clinical efficacy,and assess long-term safety,including potential epigenetic risks.
文摘The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoingin vitro fertilization (IVF)has been a debated hot topic among reproductive specialists. Each approach presents distinct advantages and disadvantages,with fresh sperm typically showing superior sperm quality, while frozen sperm offers logistical flexibility and a reliable backup forrepeated cycles. This review summarizes the latest advancements in sperm retrieval and cryopreservation techniques, providingpractitioners with a comprehensive analysis of each option’s strengths and limitations. Comparative studies indicate that, althoughfresh sperm often has better quality metrics, cryopreservation methods such as vitrification have significantly improved postthawoutcomes, making frozen sperm a viable choice in assisted reproductive technologies (ART). The findings show comparablerates for fertilization, implantation, clinical pregnancy, and live birth between fresh and frozen microdissection testicular spermextraction (micro-TESE) sperm in many cases, although patient-specific factors such as timing, cost-effectiveness, and proceduralconvenience should guide the final decision. Ultimately, the choice of using fresh or frozen sperm should align with the individualneeds and conditions of patients. This tailored approach, supported by the latest advancements, can optimize ART outcomes andprovide personalized reproductive care.
文摘Azoospermia is characterized by the absence of sperm in the ejaculate and is categorized into obstructive azoospermia(OA)and nonobstructive azoospermia(NOA).For men with NOA,testicular sperm extraction(TESE)is the only method to obtain sperm for assisted reproductive technology(ART).Given the rarity of these sperm and the unpredictable success of subsequent retrieval attempts,cryopreservation of microdissection-TESE-obtained sperm is essential.Effective cryopreservation prevents the need for repeated surgical procedures and supports future ART attempts.After first delving into the physiological and molecular aspects of sperm cryopreservation,this review aims to examine the current methods and devices for preserving small numbers of sperm.It presents conventional freezing and vitrification techniques,evaluating their respective strengths and limitations in effectively preserving rare sperm,and compares the efficacy of using fresh versus cryopreserved testicular sperm.
基金supported by the China Postdoctoral Science Foundation Grant(2023M734294)Jiangsu Provincial Medical Key Discipline Cultivation Unit(JSDW202215)the National Natural Science Foundation of China(No.82001618).
文摘Primary ciliary dyskinesia(PCD)is a clinically rare,genetically and phenotypically heterogeneous condition characterized by chronic respiratory tract infections,male infertility,tympanitis,and laterality abnormalities.PCD is typically resulted from variants in genes encoding assembly or structural proteins that are indispensable for the movement of motile cilia.Here,we identified a novel nonsense mutation,c.466G>T,in cilia-and flagella-associated protein 300(CFAP300)resulting in a stop codon(p.Glu156*)through whole-exome sequencing(WES).The proband had a PCD phenotype with laterality defects and immotile sperm flagella displaying a combined loss of the inner dynein arm(IDA)and outer dynein arm(ODA).Bioinformatic programs predicted that the mutation is deleterious.Successful pregnancy was achieved through intracytoplasmic sperm injection(ICSI).Our results expand the spectrum of CFAP300 variants in PCD and provide reproductive guidance for infertile couples suffering from PCD caused by them.
基金funded by grants from the Biological Breeding-National Science and Technology Major Projects(grant number 2023ZD0405104)Inner Mongolia Education Department Special Research Project For First Class Disciplines(grant number YLXKZX-NND-007)+1 种基金Natural Science Foundation of Inner Mongolia Autonomous Region(grant number 2023MS03001)the 12th Inner Mongolia"Grassland Talent"High-level Talent Training Project(2023).
文摘Background Freezing-induced sperm damage,often associated with oxidative stress,can result in regulated cell death.Given that oxidative stress can trigger various forms of regulated cell death,the prevailing form during sperm cryopreservation remains unknown.Our study aimed to investigate this issue using cashmere goats as a model.Results We found a significant increase in lyso-phospholipids in frozen-thawed sperm suggested ferroptosis.Assessment of cryopreserved sperm,with or without prior treatment with ferroptosis or apoptosis inhibitors,demonstrated the significant efficacy of ferroptosis inhibitors in reducing freezing damage.This implicates ferroptosis as the primary form of regulated cell death induced during sperm cryopreservation.Additionally,the positive rate of transferrin receptor protein 1 was significantly lower in fresh live sperm(47.8%)than in thawed live sperm(71.5%),and the latter rate was lower than that in dead sperm(82.5%).By contrast,cleaved caspase-3 positivity showed no significant difference between fresh live sperm and thawed live sperm but was notably lower in thawed live sperm than in dead sperm.Conclusions Our findings establish ferroptosis as the dominant regulated cell death form during goat sperm cryopreservation,providing novel insights into freezing-induced sperm damage mechanisms.These findings have significant implications for optimizing cryopreservation protocols and enhancing sperm viability after freezing-thawing.
文摘Total or severe teratospermia affects the prognosis of fertility and causes serious problems for patients undergoing assisted reproduction[1].The pathophysiological mechanism of teratospermia is unclear.It has been shown that patients with sperm parameters abnormalities and abnormal morphology have a high rate of fragmentation and sperm DNA decondensation[2,3],and that sperm DNA fragmentation analysis could be used as a predictor factor of fertility potential[4].
文摘The analysis of the ejaculate,better known as spermiogram,represents the first and main step to identify whether a series of sperm quality parameters are within the norm and therefore are consistent with normal sperm fertilizing capacity.Among these,sperm concentration and motility are the first parameters to be evaluated through an estimation carried out by expert examiners.
文摘Depression currently affects about 280 million people worldwide and its prevalence has been increasing dramatically,especially among the young and people of reproductive age,which consequently leads to an increase in antidepressant consumption.Antidepressants are associated with sexual dysfunction in both men and women;however,their role in male fertility has been scarcely studied.Fluoxetine and sertraline,two serotonin reuptake inhibitors(SSRIs),are among the most prescribed antidepressants worldwide.To determine their possible effects,human sperm cells were exposed to either sertraline or fluoxetine at concentrations previously found in blood and seminal fluid of patients undergoing treatment.Spermatozoa were incubated for up to 24 h at 37℃ and 5%CO_(2),and important functional parameters such as sperm motility,viability,mitochondrial membrane potential,cellular reactive oxygen species(ROS)production,chromatin/DNA integrity,acrosome status,and tyrosine phosphorylation were assessed.At low levels,fluoxetine consistently decreased progressive motility throughout time while promoting fluctuations in ROS levels and sperm capacitation.Nevertheless,it did not affect viability,mitochondrial membrane potential,acrosome reaction nor chromatin/DNA integrity.Sertraline,on the other hand,had little to nonsignificant impact at low doses,but affected almost all tested parameters at supratherapeutic concentrations.Altogether,our results suggest that both antidepressants may impair sperm function,possibly through different mechanisms of action,but fluoxetine is the only exhibiting mild negative effects at doses found in vivo.
文摘The advent of intracytoplasmic sperm injection,along with the realization that many men with azoospermia due to primary testicular failure may have a few spermatozoa in their testes,has resulted in the revolutionary possibility of azoospermic men fathering their own genetic offspring.
文摘High levels of sperm DNA fragmentation(SDF)are associated with reduced assisted reproductive technology(ART)outcomes.Currently,SDF is not included in routine clinical assessment of male partners of infertile couples,but the 6th edition of the World Health Organization(WHO)manual for semen analysis included the SDF assessment in the chapter on extended semen examinations.
基金supported by The Canadian Institutes of Health Research(PJT165962).
文摘Reactive oxygen species(Ros)play a dual role in mammalian spermatozoa.At high levels,they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein,lipids,and sperm DNA.This oxidative damage is associated with male infertility.On the other hand,when RoS are produced at low levels,they participate in the redox signaling necessary for sperm capacitation.Capacitation-associated RoS are produced by the sperm oxidase,whose identity is still elusive,located in the plasma membrane of the spermatozoon.Ros,such as superoxide anion,hydrogen peroxide,nitric oxide,and peroxynitrite,activate protein kinases and inactivate protein phosphatases with the net increase of specific phosphorylation events.Peroxiredoxins(PRDXs),antioxidant enzymes that fight against oxidative stress,regulate redox signaling during capacitation.Among them,PRDX6,which possesses peroxidase and calcium-independent phospholipase A,(iPLA,)activities,is the primary regulator of redox signaling and the antioxidant response in human spermatozoa.The lysophosphatidic acid signaling is essential to maintain sperm viability by activating the phosphatidylinositol 3-kinase/protein kinase(PI3K/AKT)pathway,and it is regulated by PRDX6 iPLA2,protein kinase C(PKC),and receptor-type protein tyrosine kinase.The understanding of redox signaling is crucial to pave theway fornovel diagnostic tools and treatments of male infertility.
文摘Kamil Gill,PhD,Kamil Gill is a biologist who graduated from the University of Szczecin(Poland)with a Master degree.Dr.Gill earned his scientific degree from the Pomeranian Medical University in Szczecin(PUM).His doctoral thesis was entitled:"Molecular markers of male infertility-evaluation of selected human sperm chromatin parameters and its influence on the zygote and embryo development in assisted reproduction conditions".
文摘Except in cases of hypogonadotropic hypogonadism,the use of medical therapy before microsurgical testicular sperm extraction(micro-TESE)is controversial.In some studies,hormone therapy has been shown to improve the possibility of sperm retrieval during micro-TESE and even lead to the presence of sperm in the ejaculate in some cases,thereby obviating the need for micro-TESE.However,their routine use before micro-TESE in cases of nonobstructive azoospermia(NOA)being associated with hypergonadotropic hypogonadism and eugonadism(normogonadotropic condition)has not been supported with robust evidence.In this review,we discuss different types of medical therapy used before micro-TESE for NOA,their risks and benefits,and the available evidence surrounding their use in this setting.
