scholarly journals Molecular Profiling of Spermatozoa Reveals Correlations between Morphology and Gene Expression: A Novel Biomarker Panel for Male Infertility

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Nino Guy Cassuto ◽  
David Piquemal ◽  
Florence Boitrelle ◽  
Lionel Larue ◽  
Nathalie Lédée ◽  
...  
Keyword(s):  
Gene Expression ◽  
Male Infertility ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Poor Quality ◽  
Semen Parameters ◽  
Functional Evaluation ◽  
Sperm Analysis ◽  
Biomarker Panel ◽  
Network Analyses

Choosing spermatozoa with an optimum fertilizing potential is one of the major challenges in assisted reproductive technologies (ART). This selection is mainly based on semen parameters, but the addition of molecular approaches could allow a more functional evaluation. To this aim, we used sixteen fresh sperm samples from patients undergoing ART for male infertility and classified them in the high- and poor-quality groups, on the basis of their morphology at high magnification. Then, using a DNA sequencing method, we analyzed the spermatozoa methylome to identify genes that were differentially methylated. By Gene Ontology and protein–protein interaction network analyses, we defined candidate genes mainly implicated in cell motility, calcium reabsorption, and signaling pathways as well as transmembrane transport. RT-qPCR of high- and poor-quality sperm samples allowed showing that the expression of some genes, such as AURKA, HDAC4, CFAP46, SPATA18, CACNA1C, CACNA1H, CARHSP1, CCDC60, DNAH2, and CDC88B, have different expression levels according to sperm morphology. In conclusion, the present study shows a strong correlation between morphology and gene expression in the spermatozoa and provides a biomarker panel for sperm analysis during ART and a new tool to explore male infertility.

P–050 The effectiveness of the platelet-rich plasma treatment of men with severe oligoasthenoteratozoospermia

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
O Somova ◽  
H Ivanova ◽  
N Sotnyk ◽  
K Kovalenko ◽  
I Feskova
Keyword(s):  
Male Infertility ◽  
Sperm Motility ◽  
Testosterone Level ◽  
Assisted Reproductive Technologies ◽  
Platelet Rich Plasma ◽  
Sperm Concentration ◽  
Free Testosterone ◽  
Reproductive Technologies ◽  
Positive Effects ◽  
Free Testosterone Level

Abstract Study question To evaluate the effect of platelet-rich plasma (PRP) testicular injections on spermogram parameters of men with severe oligoasthenoteratozoospermia (OAT). Summary answer The PRP testicular injections have beneficial effects on spermatogenesis and enhance sperm concentration and motility in infertile men with OAT. What is known already The use of PRP therapy in assisted reproductive technologies is debatable. Despite the recent evidence of its positive effects in promoting endometrial and follicular growth, data from clinical studies are limited. There are only a few papers on the effectiveness of PRP therapy in the treatment of male infertility and sexual dysfunction. In more detail, the influence of PRP on spermatogenesis was carried out only on experimental animals. Although the mechanisms of its action have not yet been clarified, it is assumed that PRP, containing many biologically active molecules, realizes its effect through the tissue regeneration and cell proliferation. Study design, size, duration This prospective study included 68 men (34.6±5.2) years old with severe OAT (≤4 million/ml, motility ≤30%, normal sperm morphology ≤1%) receiving hormonal and antioxidant (AO) therapy during 6 months before in vitro fertilization cycles. 33 of them were injected once with autologous PRP (0.5 ml in each testicle). Spermogram and testosterone level were analyzed before the treatment and in 3, 4 and 6 months after it. Participants/materials, setting, methods: Sperm concentration, motility and morphology in ejaculate of 33 men of PRP group were compared with those in the group of 35 men without PRP within 6 months of starting the treatment. Total and free testosterone level were measured in blood serum. PRP was prepared by centrifuging the patient’s own blood in the anticoagulant-containing tubes. The final concentration of platelets in the obtained sample was 950.000 – 1.250 000 cells in 1 ml. Main results and the role of chance 4 months after the PRP injection, sperm concentration and motility increased in 18 of 33 men of the PRP group compared with the baseline (before the treatment) – 4.2 (1.0; 6.9) vs 1.4 (0.1; 3.4) mln/ml (p < 0.05) and 36.7 (30.6; 45.8) vs 17.7 (6.7; 28.2)% respectively (p < 0.05).The maximum increase in sperm motility (but not in sperm concentration!) was observed in 24 men in 6 months – 49.6 (39.6; 56.4)% (p < 0.05). Percent of morphologically normal spermatozoa in ejaculate slightly increased only in 12 men in that time period from 0–1% to 1–2%. The total testosterone level was 2.4 times higher than the baseline (31.6±7.2 vs 13.2±4.3 nmol/l, p < 0.05), the free testosterone level was 1.8 times higher (14.5±3.5 vs 7.9±3.0 pgl/ml, p < 0.05). Unlike the PRP group, in the group of men without PRP treatment, the sperm parameters did not changed compared with the baseline in 4 months after the starting hormonal and AO treatment. A significant increase of sperin concentration was observed only in 17 of 35 patients in 6 months. Sperm motility and percent of morphologically normal spermatozoa after the treatment did not differ from the baseline. Changes in the testosterone levels were similar to changes in PRP group. Limitations, reasons for caution Only young and middle-aged men were considered in the study. Large randomized controlled studies are required to confirm the PRP therapy efficacy and safety of f various fertility disorders. There are also no standardized protocols for PRP preparation. Wider implications of the findings: PRP therapy may have great potential for the treatment of male infertility and improving spermatogenesis. Optimization of methods of PRP preparation and dosage of testicular injections can enhance reproductive outcomes in assisted reproductive technologies. Trial registration number Not applicable

154 A method of oviductal semen deposition for use in the goat

2020 ◽  
Vol 32 (2) ◽  
pp. 203
Author(s):  
N. Buzzell ◽  
S. Blash ◽  
K. Miner ◽  
M. Schofield ◽  
J. Pollock ◽  
...  
Keyword(s):  
Assisted Reproductive Technologies ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
The United States ◽  
Reproductive Technologies ◽  
Poor Quality ◽  
Implant Removal ◽  
Suitable Alternative ◽  
Midline Laparotomy ◽  
Post Surgery

The objective of this study was to investigate a method of oviducal semen deposition as a strategy for producing offspring from poor-quality cryopreserved goat sperm. Invitro fertilisation (IVF) and AI are common assisted reproductive technologies used in small ruminants, but they have varied results in the goat. The use of poor-quality cryopreserved-thawed sperm (<50% live/dead ratio at post-thaw) can decrease the rate of success. These procedures were performed in the month of November in Central Massachusetts in the United States (42° N). Seven 10-year-old dairy goats (Saanen, Toggenburg, and Alpine breeds) were synchronised and superovulated using a progesterone implant on Day 0, a prostaglandin injection at Day 7, two daily injections of 36mg of FSH ~12h apart on Days 12-15, and progesterone implant removal on Day 14 followed by an injection of 50µg of gonadotrophin-releasing hormone. Sperm deposition was performed on Day 17 (72 h after implant removal). The animals were anaesthetised using a standardised protocol, intubated, and maintained using isoflurane, and sterile prep was performed before a midline laparotomy procedure. Straws from a single ejaculate from a transgenic founder that was cryopreserved using a commercial two-step glycerol-egg yolk-based extender were used. A straw from this collection was post-thawed 30 days after collection and, using a commercial live/dead stain, 67% live sperm was determined. The optimal type of sperm prep and sperm concentration is unknown and may be dependent on sperm quality. Therefore, different gradient preps using Vitrolife SpermGrad at three volumes (1.5 (used on two animals), 1.0, and 0.5mL) as well as two volumes of IVF Bioscience Bovine BO-SemenPrep (4.0mL (used on two animals) and 2.0mL) were used. All five pellets were diluted in 1.0mL of IVF Bioscience Bovine BO-IVF media. Sperm concentrations ranging from 75×106 to 27×106 spermmL−1 were deposited into one oviduct; then, a 10:1 dilution was performed and 7.5×106 to 2.7×10 spermmL−1 were deposited into the contralateral oviduct. The depositions were performed just proximal to the uterotubal junction in a volume of 0.1mL of diluent via a tuberculin syringe attached to a 20-gauge needle. Two days following the procedure, oviducts were flushed postmortem from three of the seven randomly selected goats. All three had fertilised embryos, and nineteen 8-cell embryos were retrieved. Three of these embryos were surgically transferred to the distal uterine horn of a suitable recipient. The recipient became pregnant and produced a single offspring. The remaining four of seven goats were killed 41 days post-surgery. Two of the four goats were pregnant, with one carrying one fetus and the other carrying five fetuses. Further studies are needed to optimise this method, but these initial results indicate that oviducal semen deposition directly into the oviduct proximal to the uterotubal junction may be a suitable alternative for producing offspring from suboptimal cryopreserved-thawed goat sperm.

scholarly journals From egg to embryo: a peripatetic journey

Reproduction ◽  
2005 ◽  
Vol 130 (6) ◽  
pp. 825-828 ◽  
Author(s):  
Richard M Schultz
Keyword(s):  
Gene Expression ◽  
Assisted Reproductive Technologies ◽  
Egg Quality ◽  
Reproductive Technologies ◽  
Molecular Signature ◽  
High Quality ◽  
Long Term Consequences ◽  
The Uk ◽  
Effect Gene

The recent surge of interest in oocyte development has been spurred in large part by the increasing implementation of assisted reproductive technologies (ART) to treat human infertility. What is becoming apparent is that ‘egg quality’ is a primary factor in the success of ART (Sauer 1998), and yet we know virtually nothing about the molecular signature of a ‘high quality’ oocyte, i.e., an oocyte that is capable of maturing, being fertilized and supporting development to term. We are gaining marked insights, however, into how sperm activate eggs and the changes in gene expression that accompany preimplantation development. Nevertheless, embryo culture is known to effect gene expression (Rinaudo & Schultz 2004), the long-term consequences of which are only recently being unmasked. This review will briefly highlight these topics that were presented during the Biennial Joint Meeting of the UK Fertility Societies at Warwick University in April 2005.

Ex vivo early embryo development and effects on gene expression and imprinting

2005 ◽  
Vol 17 (3) ◽  
pp. 361 ◽  
Author(s):  
David K. Gardner ◽  
Michelle Lane
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
Assisted Reproductive Technologies ◽  
Ex Vivo ◽  
Reproductive Technologies ◽  
Developmental Competence ◽  
Laboratory Animals ◽  
Cell Physiology ◽  
Mammalian Embryo

The environment to which the mammalian embryo is exposed during the preimplantation period of development has a profound effect on the physiology and viability of the conceptus. It has been demonstrated that conditions that alter gene expression, and in some instances the imprinting status of specific genes, have all previously been shown to adversely affect cell physiology. Thus, questions are raised regarding the aetiology of abnormal gene expression and altered imprinting patterns, and whether problems can be averted by using more physiological culture conditions. It is also of note that the sensitivity of the embryo to its surroundings decreases as development proceeds. Post compaction, environmental conditions have a lesser effect on gene function. This, therefore, has implications regarding the conditions used for IVF and the culture of the cleavage stage embryo. The developmental competence of the oocyte also impacts gene expression in the embryo, and therefore superovulation has been implicated in abnormal methylation and imprinting in the resultant embryo. Furthermore, the genetics and dietary status of the mother have a profound impact on embryo development and gene expression. The significance of specific animal models for human assisted reproductive technologies (ART) is questioned, given that most cattle data have been obtained from in vitro-matured oocytes and that genes imprinted in domestic and laboratory animals are not necessarily imprinted in the human. Patients treated with ART have fertility problems, which in turn may predispose their gametes or embryos to greater sensitivities to the process of ART. Whether this is from the drugs involved in the ovulation induction or from the IVF, intracytoplasmic sperm injection or culture procedures themselves remains to be determined. Alternatively, it may be that epigenetic alterations are associated with infertility and symptoms are subsequently revealed through ART. Whatever the aetiology, continued long-term monitoring of the children conceived through ART is warranted.

P-050 The effectiveness of the platelet-rich plasma treatment of men with severe oligoasthenoteratozoospermia

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
O Somova ◽  
H Ivanova ◽  
N Sotnyk ◽  
K Kovalenko ◽  
I Feskova
Keyword(s):  
Male Infertility ◽  
Sperm Motility ◽  
Testosterone Level ◽  
Assisted Reproductive Technologies ◽  
Platelet Rich Plasma ◽  
Sperm Concentration ◽  
Free Testosterone ◽  
Reproductive Technologies ◽  
Positive Effects ◽  
Free Testosterone Level

Abstract Study question To evaluate the effect of platelet-rich plasma (PRP) testicular injections on spermogram parameters of men with severe oligoasthenoteratozoospermia (OAT). Summary answer The PRP testicular injections have beneficial effects on spermatogenesis and enhance sperm concentration and motility in infertile men with OAT. What is known already The use of PRP therapy in assisted reproductive technologies is debatable. Despite the recent evidence of its positive effects in promoting endometrial and follicular growth, data from clinical studies are limited. There are only a few papers on the effectiveness of PRP therapy in the treatment of male infertility and sexual dysfunction. In more detail, the influence of PRP on spermatogenesis was carried out only on experimental animals. Although the mechanisms of its action have not yet been clarified, it is assumed that PRP, containing many biologically active molecules, realizes its effect through the tissue regeneration and cell proliferation. Study design, size, duration This prospective study included 68 men (34.6±5.2) years old with severe OAT (≤4 million/ml, motility ≤30%, normal sperm morphology ≤1%) receiving hormonal and antioxidant (AO) therapy during 6 months before in vitro fertilization cycles. 33 of them were injected once with autologous PRP (0.5 ml in each testicle). Spermogram and testosterone level were analyzed before the treatment and in 3, 4 and 6 months after it. Participants/materials, setting, methods Sperm concentration, motility and morphology in ejaculate of 33 men of PRP group were compared with those in the group of 35 men without PRP within 6 months of starting the treatment. Total and free testosterone level were measured in blood serum. PRP was prepared by centrifuging the patient’s own blood in the anticoagulant-containing tubes. The final concentration of platelets in the obtained sample was 950.000 – 1.250 000 cells in 1 ml. Main results and the role of chance 4 months after the PRP injection, sperm concentration and motility increased in 18 of 33 men of the PRP group compared with the baseline (before the treatment) – 4.2 (1.0;6.9) vs 1.4 (0.1;3.4) mln/ml (p < 0.05) and 36.7 (30.6;45.8) vs 17.7 (6.7;28.2) % respectively (p < 0.05).The maximum increase in sperm motility (but not in sperm concentration!) was observed in 24 men in 6 months – 49.6 (39.6;56.4) % (p < 0.05). Percent of morphologically normal spermatozoa in ejaculate slightly increased only in 12 men in that time period from 0-1 % to 1-2%. The total testosterone level was 2.4 times higher than the baseline (31.6±7.2 vs 13.2±4.3 nmol/l, p < 0.05), the free testosterone level was 1.8 times higher (14.5±3.5 vs 7.9±3.0 pgl/ml, p < 0.05). Unlike the PRP group, in the group of men without PRP treatment, the sperm parameters did not changed compared with the baseline in 4 months after the starting hormonal and AO treatment. A significant increase of sperin concentration was observed only in 17 of 35 patients in 6 months. Sperm motility and percent of morphologically normal spermatozoa after the treatment did not differ from the baseline. Changes in the testosterone levels were similar to changes in PRP group. Limitations, reasons for caution Only young and middle-aged men were considered in the study. Large randomized controlled studies are required to confirm the PRP therapy efficacy and safety of f various fertility disorders. There are also no standardized protocols for PRP preparation. Wider implications of the findings PRP therapy may have great potential for the treatment of male infertility and improving spermatogenesis. Optimization of methods of PRP preparation and dosage of testicular injections can enhance reproductive outcomes in assisted reproductive technologies. Trial registration number not applicable

The effect of the breeding season, cryopreservation and physiological extender on selected sperm and semen parameters of four ferret species: implications for captive breeding in the endangered black-footed ferret

2009 ◽  
Vol 21 (2) ◽  
pp. 351 ◽  
Author(s):  
G. van der Horst ◽  
R. M. Kitchin ◽  
M. van der Horst ◽  
R. W. Atherton
Keyword(s):  
Breeding Season ◽  
Sperm Motility ◽  
Captive Breeding ◽  
Assisted Reproductive Technologies ◽  
Sperm Quality ◽  
Reproductive Technologies ◽  
Semen Parameters ◽  
Breeding Cycle ◽  
Sperm Characteristics ◽  
Motion Characteristics

In the present investigation, comparative baseline information on selected sperm characteristics of ejaculate spermatozoa of the domestic (Mustela putorius furo), fitch (Mustela sp.) and black-footed ferrets (Mustela nigripes) and the Siberian polecat (Mustela eversmanni) are presented. The main emphasis was to establish differences and similarities among these species in relation to semen and sperm quality during the breeding season, in cryopreservation success and in supporting sperm motility in different extenders or physiological media. The results confirm that most sperm morphology abnormalities were evident during the beginning of the breeding cycle in all four species. No significant interspecies differences were apparent in the sperm attributes examined, for all sampling months during the breeding season. Moreover, all species exhibited comparable patterns of reproductive seasonality. Cryopreservation suppressed sperm characteristics equally in all species studied. Ejaculate spermatozoa of closely related ferret species shared many similar motion characteristics using computer-aided sperm motility analysis. These results suggest that the basic sperm physiology of the ferret species under examination is very similar. Disparate to the interspecies comparisons, there were significant differences for most sperm motion parameters when spermatozoa of any of the ferrets were compared in different extenders. Assisted reproductive technologies developed for use in domestic ferret, fitch ferret or Siberian polecat may be successfully applied to captive breeding of the black-footed ferret using semen during any of the functional breeding months.

scholarly journals Clinical significance of sperm morphology in male infertility treatment

2021 ◽  
Vol 14 (3) ◽  
pp. 127-132
Author(s):  
Yu.V. Olefir ◽  
◽  
D.M. Monakov ◽  
◽  
Keyword(s):  
Male Infertility ◽  
Clinical Significance ◽  
Sperm Morphology ◽  
Assisted Reproductive Technologies ◽  
Intrauterine Insemination ◽  
Reproductive Technologies ◽  
Vitro Fertilization ◽  
Normal Sperm ◽  
Starting Point

Introduction. Sperm morphology is one of the most common tests in fertility practice. The interlaboratory variability is the main drawback of the method. The clinical significance of sperm morphology in assisted reproductive technologies is controversial. The aim of the review is to address this question. Materials and methods. The search of relevant publications was carried out in PubMed and e-Library databases using the keywords «male infertility», «sperm morphology», «teratozoospermia», «IUI», «IVF», «ICSI». Conference abstracts and dissertations were excluded from analysis and 56 publications were included in this literature review. Results. The small numbers of studies were evaluated to the effect of teratozoospermia on the likelihood of natural pregnancy. The pregnancy rate was higher in the group of couples with normozoospermia. In the couples with severe teratozoospermia pregnancies rate was also detected. The most studies did not reveal a statistically significant effect of tertozoospermia on the frequency of pregnancy during intrauterine insemination. The data about the influence of sperm morphology on in vitro fertilization are contradictory. Early studies showed a positive correlation between normal sperm morphology and frequency of conception, but these results were not confirmed in further studies. The most studies have not been revealed the correlation between normal sperm morphology male fertility status, clinical and live birth rate. Discussion. To date the spermatozoa mofophology studying remains the «starting point» ofa man's examination for infertility. However, the data available do not confirm its role in choosing the method ofassisted reproductive technologies or predicting their results. Conclusions. To date the role of sperm morphology on conceive and pregnancy frequencies in ART use is controversal. The data available does not confirm the value of this test as a proxy of higher pregnancy and birth of healthy child probability. The further studies are required to address this question.

scholarly journals Is there a connection with basic sperm parameters and age?

2020 ◽  
Vol 13 (4) ◽  
pp. 58-64
Author(s):  
A.I. Ryzhkov ◽  
◽  
I.S. Shormanov ◽  
S.Yu. Sokolova ◽  
◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Male Fertility ◽  
Reproductive Technologies ◽  
Semen Parameters ◽  
Sperm Dna Fragmentation ◽  
Who Guidelines ◽  
Negative Relation ◽  
Sperm Analysis ◽  
Mobile Forms ◽  
Sperm Dna

Introduction. TStandard sperm examination, performed according to WHO guidelines, remains the main method of assessment of male fertility. At the same time, this research method has a number of significant limitations, including a low predictive value regarding the outcomes of assisted reproductive technologies (ART) programs and the outcome of naturally occurring pregnancies. The limitations of standard sperm examination dictate the need for additional methods for assessing male fertility. The most promising and widely used test is the assessment of the level of sperm DNA fragmentation. Aim. To evaluate the associations between the levels of sperm DNA fragmentation and the age of the patients along with the following parameters used as a part of standard sperm analysis: semen volume, total number of spermatozoa, percentages of progressively-mobile, non-progressively-mobile and immobile forms, percentages of morphologically normal forms and the forms bearing head defects within the structure of total number of morphological anomalies, as well as semen leukocyte count. Materials and methods. Study materials were the examination results from 121 males aged from 21 to 53 years old (mean age 32.7±4.5 years old), undergoing an examination within the Clinical Institution «Mother and Child – Yaroslavl» during a time period from January 2019 until April 2020. Standard sperm analysis procedures were carried out according to latest edition of WHO Guidelines (2010). The determinations of germ cell DNA fragmentation levels were performed using the TUNEL method. Results. The test results have revealed a weak negative relation between the sperm DNA fragmentation (%) and the percentage of progressively-mobile forms (%) – r = -0.26 (p <0.01). The correlation of sperm DNA fragmentation with age and other parameters was considered statistically insignificant. The second stage of the analysis have demonstrated that the increased degree sperm DNA fragmentation is 1.8-fold more often found in the patients having signs of astenozoospermia (23.6%) comparing to the patients showing normal degree of spermatozoa mobility (13.1%), (p <0.05). Conclusions. The level of sperm DNA fragmentation correlates with the percentage of progressively-mobile forms of spermatozoa (negative relation) and no correlations were found with other semen parameters and with the age. The rates of increased levels of sperm DNA fragmentation are 1.8-fold more often found in astenozoospermia patients comparing to the patients showing normal degrees of spermatozoa mobility.

scholarly journals FEATURES OF LEGAL REGULATION OF GENOMIC RESEARCHES: FAILURES, SECURITY CONCERNS AND PROSPECTS

2020 ◽  
Vol 30 (1) ◽  
pp. 145-152
Author(s):  
A.A. Mokhov ◽  
A.N. Yavorsky ◽  
A.R. Pozdeev
Keyword(s):  
Russian Federation ◽  
Assisted Reproductive Technologies ◽  
Genetic Diseases ◽  
Reproductive Technologies ◽  
Legal Regulation ◽  
Poor Quality ◽  
Federal Law ◽  
The Russian Federation ◽  
Public Health Protection ◽  
Genetic Consultation

The urgency of the article is determined by the need to ensure biological and ecological safety of Russia. The article gives examples of failures of auxiliary reproductive technologies in the Russian Federation and abroad which at present do not have adequate algorithms of legal resolution and are unsafe for ecology and man. Investigating the existing Russian legislation it has been revealed that passing only of obligatory medico-genetic consultation and incomplete medico-genetic inspection with approximate assessment of the genetic status of the donor does not exclude development of the fetus with severe genetic diseases, which is considered by the consumer of the service as poor quality and unsafe. It is noted that in cases of use of biomaterials from abroad, a comparative study of the genome becomes unlikely due to paragraph 7 of Art. 55 of the Federal Law of 21.11.2011 № 323-FZ "On the fundamentals of public health protection in the Russian Federation" and the order of the Ministry of Health of Russia from 30.08.2012. (ed. 01.02.2018) № 107n "On the order of using assisted reproductive technologies, contraindications and restrictions on their use". The conclusion was drawn on the need to develop and fix organizational and legal mechanisms and develop appropriate procedures.

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