scholarly journals Sperm Cohort-Specific Zinc Signature Acquisition and Capacitation-Induced Zinc Flux Regulate Sperm-Oviduct and Sperm-Zona Pellucida Interactions

2020 ◽  
Vol 21 (6) ◽  
pp. 2121 ◽  
Author(s):  
Karl Kerns ◽  
Momal Sharif ◽  
Michal Zigo ◽  
Wei Xu ◽  
Lauren E. Hamilton ◽  
...  
Keyword(s):  
Zona Pellucida ◽  
Semen Analysis ◽  
Inhibitory Effect ◽  
26S Proteasome ◽  
Matrix Metalloproteinase 2 ◽  
Zinc Ions ◽  
New Paradigm ◽  
Male Factor ◽  
Sperm Penetration

Building on our recent discovery of the zinc signature phenomenon present in boar, bull, and human spermatozoa, we have further characterized the role of zinc ions in the spermatozoa’s pathway to fertilization. In boar, the zinc signature differed between the three major boar ejaculate fractions, the initial pre-rich, the sperm-rich, and the post-sperm-rich fraction. These differences set in the sperm ejaculatory sequence establish two major sperm cohorts with marked differences in their sperm capacitation progress. On the subcellular level, we show that the capacitation-induced Zn-ion efflux allows for sperm release from oviductal glycans as analyzed with the oviductal epithelium mimicking glycan binding assay. Sperm zinc efflux also activates zinc-containing enzymes and proteases involved in sperm penetration of the zona pellucida, such as the inner acrosomal membrane matrix metalloproteinase 2 (MMP2). Both MMP2 and the 26S proteasome showed severely reduced activity in the presence of zinc ions, through studies using by gel zymography and the fluorogenic substrates, respectively. In the context of the fertilization-induced oocyte zinc spark and the ensuing oocyte-issued polyspermy-blocking zinc shield, the inhibitory effect of zinc on sperm-borne enzymes may contribute to the fast block of polyspermy. Altogether, our findings establish a new paradigm on the role of zinc ions in sperm function and pave the way for the optimization of animal semen analysis, artificial insemination (AI), and human male-factor infertility diagnostics.

scholarly journals Role of Laparoscopy in Infertility : Review Article

2012 ◽  
Vol 2 (2) ◽  
pp. 99-103 ◽  
Author(s):  
S Jahan
Keyword(s):  
Fiber Optics ◽  
Semen Analysis ◽  
Uterine Fibroids ◽  
Scar Tissue ◽  
Direct Visualization ◽  
Male Factor ◽  
Operative Laparoscopy ◽  
Pelvic Organs ◽  
Carbon Dioxide Co2

Infertility is defined as failure to conceive during one year of unprotected frequent intercourse. Leading causes of infertility include tubal disease, ovulatory disorders, uterine or cervical factors, endometriosis and male factor infertility. A laparoscope is a thin fiber optic telescope that is inserted into the abdomen usually through the belly button. The fiber optics allow a light to used to see inside the abdomen. Carbon dioxide (CO2) gas is placed into the abdomen prior to inserting the laparoscope. Generally, laparoscopy should be reserved for couples who have already completed a more basic infertility evaluation including assessing for ovulation, ovarian reserve, ultrasound and hysterosalpingogram for the female and semen analysis for the male. Laparoscopy can help physicians diagnose many gynecological problems including endometriosis, uterine fibroids and other structural abnormalities, ovarian cysts, adhesions (scar tissue), and ectopic pregnancy. Robotic assisted laparoscopic surgery (RAL) is a more recent development and a form of operative laparoscopy. In RAL, the instruments and telescope are very similar to conventional laparoscopy, but they are attached to a robot which in turn is controlled by the surgeon who is seated at a viewing console. Women who have been diagnosed with endometriosis are more likely to experience infertility, and observational studies have shown that the monthly probability of pregnancy in women with endometriosis is about half of the probability in normal women. In spite of this well-documented association, a true cause and effect relationship has not been established. Laparoscopy is used world-wide to investigate infertility. It is an essential part of full assessment and treatment of infertility. It provides direct visualization of the pelvic organs, ovarian and tubal status and can elucidate the site of tubal obstruction. It has got an advantage of direct visualization of the pelvic organs and the peri-tubal status resulting in greater information as compared to hysterosalpingography and ultrasonography. The advance in instrument technology has made this procedure more productive and less hazardous. Laparoscopy is the most dependable tool to investigate pelvic pathology. The role of laparoscopy in diagnosis of infertility both primary as well as secondary is established beyond any doubt.DOI: http://dx.doi.org/10.3329/birdem.v2i2.12324 (Birdem Med J 2012; 2(2): 99-103)

163 HYPERACTIVATION OF STALLION SPERM IN FOLLICULAR FLUID FOR IN VITRO FERTILIZATION OF EQUINE OOCYTES

2012 ◽  
Vol 24 (1) ◽  
pp. 193 ◽  
Author(s):  
A. Lange-Consiglio ◽  
F. Cremonesi
Keyword(s):  
In Vitro Fertilization ◽  
Sperm Motility ◽  
Follicular Fluid ◽  
Zona Pellucida ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Vitro Fertilization ◽  
Sperm Penetration ◽  
Low Incidence

In vitro fertilization has remained elusive in the horse, as evidenced by low sperm penetration rates when IVF has been attempted with in vivo- or in vitro-matured oocytes. It is likely that the low sperm penetration rates observed in IVF studies are due to the inability to appropriately capacitate or hyperactivate, or both, stallion sperm in the laboratory. The acquisition of hyperactivated sperm motility has been observed within the oviducts of mammals at the time of fertilization and is required for zona pellucida penetration in conjunction with the acrosome reaction (AR). Although the zona pellucida is considered the prime physiological inducer of AR, previous studies have shown a low incidence of AR in zona pellucida-bound stallion spermatozoa after 1 h of in vitro binding. This low incidence suggests that, besides the zona pellucida glycoproteins, another major factor might be responsible for AR. Protein-bound progesterone, present in equine follicular fluid (FF), has been demonstrated to induce AR in stallion spermatozoa. In this context, the aims of this study were (1) to hyperactivate stallion sperm in FF and (2) to verify whether this hyperactivation supports equine IVF. Pooled FF, aspirated from the preovulatory follicles of oestrous mares, was used and its progesterone concentration was determined by immune enzymatic assay. Spermatozoa from fertile stallions selected by a swim-up procedure were pre-incubated for 6 h in capacitating medium (modifed Whittens's medium (WM) supplemented with 25 mM NaHCO3 and 7 mg mL–1 of BSA) and then incubated for 6 h at 37°C in either FF or capacitating WM. Sperm motility was assayed by computer-assisted semen analysis, rates of AR were assessed by fluorescein isothiocyanate-PNA staining and rate of apoptosis was assessed by an annexin V test. For IVF, spermatozoa were incubated at 10 × 106 sperm mL–1 in capacitating WM for 6 h and then diluted to 1 × 106 sperm mL–1 in capacitating WM with or without 10% of FF. Five mature mare oocytes were transferred into droplets (100 μL) of the sperm suspensions covered with mineral oil and then incubated for 18 h at 38.5°C in 5% CO2 in humidified air. After that, oocytes were transferred to an embryo culture medium (DMEM/F-12) for an additional 3 days. Data were analysed by ANOVA. Treatment of sperm with FF resulted in a significant (P ≤ 0.05) decrease of 3 motility variables indicative of hyperactivation: straight line velocity, straightness and linearity. The highest rate of AR (29.44%) and a lower rate of apoptosis (16.93%) were obtained after 4 h of incubation in follicular fluid. By coupling capacitating conditions with the induction of hyperactivation using follicular fluid, we have obtained reproducible percentages of 8-cell-stage embryos (18.56%) in our IVF experiments. Conversely, sperm incubated in capacitating conditions but not treated with FF did not fertilize (0%). It is concluded that mare FF does not impair sperm viability, stimulates equine sperm hyperactivation in vitro, induces the AR and supports equine IVF.

scholarly journals Study on Transvaginal Sonographic Diagnosis of Infertility of Infertility: A Comparative Outcome With Clinical Correlations

2012 ◽  
Vol 18 (Number 1) ◽  
pp. 9-19
Author(s):  
T Begum ◽  
F Begum ◽  
Md. N Islam
Keyword(s):  
Semen Analysis ◽  
Cyst Formation ◽  
Normal Range ◽  
Male Factor ◽  
Cross Sectional ◽  
Obstetrics And Gynaecology ◽  
Clinical Correlations ◽  
Medical University ◽  
Renal Abnormalities

A cross sectional comparative study was doneon ran.cvaginal sonographic (TVS) findings to diagnose infertility in 110 patients attended theta t.patient department of Obstetrics and Gynaecology at Bangabandhu Sheikh Mt jib Medical University (BSMM1.1). The study was done with an object ro asses the role of traavvaginal ultra sonography as a sensitive detector for the diagnosis of poly cystic ovarian disease. endonteriomas. leontyomas.retroversion of itterus.pelvic inflammatory disease. adnegal mass or pelvic tumour. congenital abnormalities of paramesonephric (niillarion)ducts and also its association with renal abnormalities. The age of the study patients ranged from 19-40 yrs. They were grouped into three categories. Amongst them 58 cases was represented with primary infertility which is 47.27 % Mike total cases. In this study 37 patients was diagnosed by TVS as having PCOD amongst them II patients presented with oligomenorrhoea. Comparison between clinical correlation and TVS findings were done in which 15 patients clinically seemed to have with endometrioma with cyst formation which correlates 10 cases of TVS findings. TVS findings were normal in 21 patients. But they had come only with primary or secondary infertility. This study included only those patients Otiose husband's semen analysis is within normal range and no other male factor. These are the unexplained infertility.

14 Opening the Black Box of Fertility Prediction

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 141-142
Author(s):  
Karl Kerns
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
Precision Agriculture ◽  
Semen Analysis ◽  
Black Box ◽  
Sperm Capacitation ◽  
New Paradigm ◽  
Male Factor ◽  
Technological Advances ◽  
Standard Quality

Abstract Analysis of the U.S. swine herd shows variation in pregnancy rate is more attributable to male-factor subfertility than the dam. To date, a limited degree of correlations has been observed between conventional semen analysis parameters and actual fertility after standard quality cutoffs are met. Thus, a clear ability to predict male-factor fertility is lacking. Knowledge of what makes fertilization competent spermatozoa has been long sought after for centuries. It was only in the last half-century that we understood spermatozoa undergo a biological process after ejaculation to acquire the capacity to fertilize. Since then, work has been done to elucidate the molecular pathways involved in sperm capacitation. Recent technological advances in flow cytometry, namely image-based flow cytometry, allows for high-throughput, single-cell phenotyping. Single-cell phenotyping with biomarkers reflecting significant sperm capacitation events, mitochondrial status, cell health, and more, allows multi-million bioimage data sets to be easily attained. These datasets can then be analyzed utilizing machine and deep learning analytic methods and correlated with single sire field fertility data to open the black box of boar fertility prediction. Our findings establish a new paradigm in sperm function and pave the way for accurate fertility prediction in future precision agriculture applications. This work was supported by the National Institute of Food and Agriculture (NIFA), U.S. Department of Agriculture (USDA) award number 2019-67012-29714.

scholarly journals Human sperm bind to the N-terminal domain of ZP2 in humanized zonae pellucidae in transgenic mice

2012 ◽  
Vol 197 (7) ◽  
pp. 897-905 ◽  
Author(s):  
Boris Baibakov ◽  
Nathan A. Boggs ◽  
Belinda Yauger ◽  
Galina Baibakov ◽  
Jurrien Dean
Keyword(s):  
Zona Pellucida ◽  
Human Sperm ◽  
Binding Assays ◽  
Terminal Domain ◽  
Sperm Binding ◽  
Gamete Recognition ◽  
Recombinant Peptides ◽  
Sperm Penetration ◽  
Mouse Eggs

Fertilization requires taxon-specific gamete recognition, and human sperm do not bind to zonae pellucidae (ZP1–3) surrounding mouse eggs. Using transgenesis to replace endogenous mouse proteins with human homologues, gain-of-function sperm-binding assays were established to evaluate human gamete recognition. Human sperm bound only to zonae pellucidae containing human ZP2, either alone or coexpressed with other human zona proteins. Binding to the humanized matrix was a dominant effect that resulted in human sperm penetration of the zona pellucida and accumulation in the perivitelline space, where they were unable to fuse with mouse eggs. Using recombinant peptides, the site of gamete recognition was located to a defined domain in the N terminus of ZP2. These results provide experimental evidence for the role of ZP2 in mediating sperm binding to the zona pellucida and support a model in which human sperm–egg recognition is dependent on an N-terminal domain of ZP2, which is degraded after fertilization to provide a definitive block to polyspermy.

scholarly journals Zinc ion fluxes on the pathway to mammalian sperm fertilization competency

2019 ◽  
Author(s):  
◽  
Karl Kerns
Keyword(s):  
Semen Analysis ◽  
Round Spermatid ◽  
Zinc Ion ◽  
Sperm Capacitation ◽  
New Paradigm ◽  
Vitro Fertilization ◽  
Semen Storage ◽  
Mammalian Spermatozoa

For decades, the role of the divalent ion calcium was the focus of understanding mechanisms leading to sperm fertilization competency and management of semen storage. Little focus has been placed on other divalent ions, including zinc ion (Zn2+). Further, the ultimate maturation event preparing mammalian spermatozoa for fertilization, sperm capacitation, was first described in 1951, yet its regulatory mechanisms remain poorly understood. Here, we document a novel biological phenomenon of a unique Zn2+ distribution (further zinc signature) associated with mammalian spermatozoa from the round spermatid stage of spermiogenesis, to epididymal maturation, ejaculation, and up to 72 hours of liquid semen storage. Using image-based flow cytometry (IBFC), we identified four distinct sperm zinc signatures present in boar, bull, and human spermatozoa. The zinc signature was altered after sperm capacitation, reduced by proteasomal inhibitors, removed by zinc chelation, and maintained with addition of external ZnCl2. The zinc signature differed between the three major boar ejaculate fractions. These differences set in sperm ejaculatory sequence likely establish two major sperm cohorts; one destined for populating the sperm oviductal reservoir and the other that is capable of fertilizing mature, ovulated oocytes at the time of mating/insemination. Management of the sperm zinc signature prevented spontaneous, pathological capacitation, by day 3 of extended liquid boar semen storage and could allow for use of fewer sperm per artificial insemination (AI) dose to increase the usage of high genetic value sires. A newly formulated semen extender was able to mimic qualities of the pre-sperm rich fraction. On a subcellular level, the capacitation induced Zn2+ efflux allows for release from oviductal glycans studied with the oviductal epithelium mimicking glycan binding assay. Sperm Zn2+ efflux also activates zinc-containing enzymes involved in sperm penetration of the zona pellucida, such as the inner acrosomal membrane metalloproteinase MMP2 that had a severely reduced activity in the presence of Zn2+ by gel zymography. In context of the fertilization-induced oocyte zinc spark and the ensuing, oocyteissued polyspermy-blocking zinc shield, the inhibitory effect of Zn2+ on spermborne enzymes may contribute to the fast block of polyspermy. Altogether, our findings establish a new paradigm on the role of Zn2+ in sperm function, paving the way for improved semen analysis, in vitro fertilization (IVF) and the optimization of AI and semen distribution.

Effect of Dipyridamole on Spontaneous Platelet Aggregation in Whole Blood Decreases with the Time After Venepuncture: Evidence for the Role of ADP

1987 ◽  
Vol 58 (02) ◽  
pp. 744-748 ◽  
Author(s):  
A R Saniabadi ◽  
G D O Lowe ◽  
J C Barbenel ◽  
C D Forbes
Keyword(s):  
Platelet Aggregation ◽  
Correlation Coefficient ◽  
Whole Blood ◽  
Blood Platelet ◽  
Inhibitory Effect ◽  
Time Interval ◽  
Adp Receptor ◽  
Spontaneous Platelet Aggregation

SummarySpontaneous platelet aggregation (SPA) was studied in human whole blood at 3, 5, 10, 20, 30, 40 and 60 minutes after venepuncture. Using a whole blood platelet counter, SPA was quantified by measuring the fall in single platelet count upon rollermixing aliquots of citrated blood at 37° C. The extent of SPA increased with the time after venepuncture, with a correlation coefficient of 0.819. The inhibitory effect of dipyridamole (Dipy) on SPA was studied: (a) 10 μM at each time interval; (b) 0.5-100 μM at 3 and 30 minutes and (c) 15 μM in combination with 100 μM adenosine, 8 μM 2-chloroadenosine (2ClAd, an ADP receptor blocker) and 50 μM aspirin. There was a rapid decrease in the inhibitory effect of Dipy with the time after venepuncture; the correlation coefficient was -0.533. At all the concentrations studied, Dipy was more effective at 3 minutes than at 30 minutes after venepuncture. A combination of Dipy with adenosine, 2ClAd or aspirin was a more effective inhibitor of SPA than either drug alone. However, when 15 μM Dipy and 10 μM Ad were added together, the inhibitory effect of Dipy was not increased significantly, suggesting that Dipy inhibits platelet aggregation independent of Ad. The increase in SPA with the time after venepuncture was abolished when blood was taken directly into the anticoagulant containing 5 μM 2ClAd. It is suggested that ADP released from the red blood cells is responsible for the increased platelet aggregability with the time after venepuncture and makes a serious contribution to the artifacts of in vitro platelet function studies.

INOVASI PEMBELAJARAN AKIDAH AKHLAK MENGGUNAKAN METODE ROLE PLAY DALAM MENINGKATKAN PRESTASI BELAJAR SISWA

2018 ◽  
Vol 2 (1) ◽  
pp. 52-63
Author(s):  
Ansori Ansori
Keyword(s):  
Data Reduction ◽  
Role Play ◽  
Learning Goals ◽  
New Paradigm ◽  
Learning Materials ◽  
Analysis Technique ◽  
Flexible Schedule ◽  
Using Data ◽  
Real Action

The use of various methods will greatly help students in achieving learning goals. As role play method is one way mastery of learning materials through the development of imagination and appreciation of students on learning materials. Data collection techniques in this study are observation, interviews, and documentation. To analyze the data in this research using data analysis technique of Miles and Huberman model that is data reduction (Data Reduction), data presentation (Data Display) and conclusion (Conclution Drawing / verification) The findings in this research is innovation of role play method can change paradigm to the new paradigm so that the role of the teacher is more as a facilitator, counselor, consultant, and comrade study Flexible schedule, open as needed Learning directed by students themselves Problem-based, project, real world, real action, and reflection Design and investigation. Computers as tools, and dynamic media presentations.

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