Transcriptome profile and association study revealed STAT3 gene as a potential quality marker of bovine gametes

Zygote ◽  
2020 ◽  
Vol 28 (2) ◽  
pp. 116-130
Author(s):  
Nasser Ghanem ◽  
Dessie Salilew-Wondim ◽  
Michael Hoelker ◽  
Karl Schellander ◽  
Dawit Tesfaye

SummaryThe present study was aimed to investigate differences in molecular signatures in oocytes derived from Holstein-Friesian heifers with different genetic merit for fertility, euthanized during day 0 or day 12 of the estrous cycle. Moreover, association between single nucleotide polymorphisms (SNPs) of ODC1 and STAT3 genes and bull fertility traits was investigated. The gene expression patterns were analyzed using cDNA array and validated with quantitative real-time polymerase chain reaction (PCR). The result revealed that several genes have shown not only to be regulated by fertility merit but also by the day of oocyte recovery during the estrous cycle. The STAT3 gene was found to be upregulated in oocytes recovered from animals with high fertility merit at both day 0 and day 12. Some other genes like PTTG1, ODC1 and TUBA1C were downregulated at day 0 and upregulated at day 12 in high, compared with low, fertility merit recovered oocytes. In contrast, the transcript abundance of TPM3 was upregulated at day 0 and downregulated at day 12 in high, compared with low, fertility merit recovered oocytes. In addition, ODC1 and STAT3 were found to be associated (P < 0.05) with sperm quality traits as well as flow cytometry parameters. Therefore, the expression of several candidate genes including ODC1 and STAT3 was related to the genetic merit of the cow. In addition polymorphisms in these two genes were found to be associated with bull semen quality.

2020 ◽  
Vol 21 (20) ◽  
pp. 7489
Author(s):  
Zijun Lu ◽  
Xiaotong Guo ◽  
Zhiyu Huang ◽  
Juan Xia ◽  
Xiang Li ◽  
...  

Autotetraploid rice is a useful rice germplasm for polyploid rice breeding. However, low fertility limits its commercial production. A neo-tetraploid rice with high fertility was developed from the progenies of crossing between autotetraploid lines by our research group. Our previous study showed that a myeloblastosis (MYB) transcription factor, MOF1, might be associated with the pollen development in tetraploid rice. However, little information is available about its role in pollen development in tetraploid rice. Here, we identified a new haplotype of MOF1 from neo-tetraploid rice and marked it as MOF1a. Transcriptome and qRT-PCR analysis demonstrated that MOF1a highly expressed in anthers, and displayed differential expression in neo-tetraploid rice compared to tetraploid rice line with low pollen fertility. The mutant (mof1a) of MOF1a, which was generated by CRISPR/Cas9 knockout in neo-tetraploid rice, showed low pollen fertility, and also exhibited abnormal tapetum and middle layer development, and defective chromosome behaviors during meiosis. A total of 13 tapetal related genes were found to be up-regulated in meiotic anthers of MOF1a compared with wild type plants by RNA-seq analysis, including CYP703A3, PTC1, and OsABCG26, which had been demonstrated to affect tapetal development. Moreover, 335 meiosis-related genes displayed differential expression patterns at same stage, including nine important meiosis-related genes, such as metallothionein OsMT1a. These results demonstrated that MOF1a plays an important role in pollen development and provides a foundation for understanding the molecular mechanism underlying MOF1a in reproduction of tetraploid rice.


2013 ◽  
Vol 25 (1) ◽  
pp. 180 ◽  
Author(s):  
D. Kumar ◽  
P. Kumar ◽  
P. Singh ◽  
S. P. Yadav ◽  
P. S. Yadav

The breeding-soundness examination is conducted to identify and select bulls with an acceptable reproductive efficiency. In buffalo, there is meagre information regarding fertility index in relation to sperm attributes so that a future breeding bull could be selected at the age of maturity. So, to predict the fertility of buffalo breeding bulls, the present study was conducted to assess the motility parameters and integrity of acrosome, plasma membrane, and DNA of cryopreserved semen of high- and low-fertile bulls. The fertility of bulls was classified on the basis of conception rates, where buffalo bulls with conception rate <35% were considered as low-fertility bulls and those with conception rate >55% were considered as high-fertility bulls. A computer-assisted semen analyser was used for motility and viability studies, whereas integrity of acrosome, plasma membrane, and DNA were assessed by Pisum sativum agglutinin–fluorescein isothiocyanate, Annexin-V/PI, and TUNEL assay kit, respectively, under the florescence microscope. At least 200 spermatozoa were evaluated from each group, and results were analysed by ANOVA. The level of significance was observed at P < 0.05. The mean (±SE) values for various motility parameters of sperm for high-fertile bulls were total motility (56.8 ± 3.2%), average path velocity (87.22 ± 1.6 µm s–1), straight linear velocity (68.93 ± 1.9 µm s–1), and curvilinear velocity (156.52 ± 4.3 µm s–1). These values were significantly higher than those of low-fertile bulls (43.8 ± 1.7%, 79.02 ± 2.4 µm s–1, 63.42 ± 1.2 µm s–1, and 142.37 ± 2.8 µm s–1, respectively). The amplitude of lateral head displacement (6.8 ± 0.07 v. 6.5 ± 0.1 µm), beat cross frequency (33.9 ± 0.4 v. 33.43 ± 0.5 Hz), straightness (79.2 ± 0.7 v. 78.7 ± 0.6%), linearity (45.5 ± 0.4 v. 45.5 ± 0.7%), and viability (71.2 ± 0.8 v. 68.9 ± 0.8%) did not differ in both groups. The average percentage of intactness of sperm acrosome of high-fertile bulls was significantly higher (81.82 ± 0.87%) than that of low-fertility bulls (76.86 ± 0.87%). Furthermore, to assess the functionality of plasma membrane of sperm, we analyzed different stages of apoptotic-like events. The percentage of apoptotic sperm differed significantly between high-fertility (15.59 ± 0.75%) and low-fertility (25.94 ± 0.5%) bulls. The percentages of early necrotic, necrotic, and viable sperm did not differ in 2 groups. The DNA integrity in high-fertility (90.24 ± 0.94%) and low-fertility (88.37 ± 0.91%) bulls was not significantly different. In conclusion, the various parameters such as average path velocity, curvilinear velocity, straight linear velocity, and total motility; acrosomal integrity; and percentage of apoptotic sperm are useful for evaluating the semen quality of a bull to reduce the risk of using poor-fertility bulls in an AI program.


Reproduction ◽  
2007 ◽  
Vol 134 (1) ◽  
pp. 19-29 ◽  
Author(s):  
Montserrat Gomendio ◽  
Aurelio F Malo ◽  
Julian Garde ◽  
Eduardo R S Roldan

Male fertility has seldom been studied in natural populations because it has been assumed that strong selection would result in uniformly high values among males, and therefore mating success has been equated with fertilisation success. In contrast, male fertility has received much attention in studies of domestic livestock, where economic benefits rely on improving productivity, and in human infertility studies, where the efficiency of treatments depends on understanding which ejaculate traits explain reproductive failures and predict success at assisted conception. Despite years of efforts, no conclusive results have been obtained, probably because such studies have focused on opposite extremes of the range with little variation: domestic livestock have often been subject to strong artificial selection for high fertility, and human patients requiring treatment have compromised fertility. Recent findings from natural populations of red deer have shown that males differ markedly in their fertility, and have revealed the degree of variation found in different semen traits, both between and within males. Fertility trials have shown that male fertility is determined mainly by sperm swimming speed and the proportion of normal sperm, when sperm numbers are kept constant. Sperm design exerts a strong influence on sperm swimming speed, with faster swimming sperm having elongated heads, shorter midpieces and a longer principal plus terminal pieces in relation to total flagellum length. Thus, the large inter-male variation in sperm design found among natural populations underlies differences in sperm swimming speed which, in turn, determine differences in male fertility rates. Secondary sexual characters are honest indicators of male fertility, so males with large and elaborated antlers have larger testes and faster swimming sperm. Testosterone does not seem to mediate the relationship between antler size and semen quality, since it is associated with sperm production, but not with sperm quality or antler size. Finally, more fertile males produce a greater proportion of sons, who will inherit the semen traits which will enhance their fertility.


Zygote ◽  
2018 ◽  
Vol 26 (5) ◽  
pp. 388-394
Author(s):  
Thanapol Nongbua ◽  
Essraa M Al-Essawe ◽  
Anders Edman ◽  
Anders Johannisson ◽  
Jane M Morrell

SummaryThe aim of this study was to investigate the effect of adding homologous or heterologous bovine seminal plasma (SP) to SP-free sperm samples before freezing on sperm quality after thawing. Ejaculates from bulls of known fertility were used as a source of SP. The SP was removed from further aliquots of the same ejaculates by colloid centrifugation to create SP-free sperm samples; the resuspended sperm pellets were treated with homologous or heterologous SP from high or low fertility bulls at 0%, 1% or 5% before freezing. After thawing, sperm quality was evaluated by computer-assisted sperm analysis and flow cytometry for membrane integrity, reactive oxygen species, chromatin structure, mitochondrial membrane potential and protein tyrosine phosphorylation. Data were analysed using Proc MIXED, SAS®. Post-hoc comparisons were adjusted for multiplicity using Tukey’s method. The addition of SP resulted in significant differences in sperm quality, namely velocity class A, Velocity Straight Line (VSL), Velocity Average Path (VAP), Velocity Curved Line (VCL), Amplitude of Lateral Head Displacement (ALH), Hyperactive (HYP), reactive oxygen species (ROS) production and % DNA fragmentation index (DFI) (P<0.05 for each). Although adding 5% homologous SP from high fertility bulls was beneficial to sperm kinematics, 5% heterologous SP from high fertility bulls had a deleterious effect on chromatin integrity and on sperm velocity. In conclusion, adding SP may have either a beneficial effect or a deleterious effect depending on the individuals involved. It might be feasible to use this method to improve sperm quality in some circumstances.


2020 ◽  
Vol 21 (5) ◽  
pp. 1902 ◽  
Author(s):  
Anna Mańkowska ◽  
Paweł Brym ◽  
Łukasz Paukszto ◽  
Jan P. Jastrzębski ◽  
Leyland Fraser

Genetic markers have been used to assess the freezability of semen. With the advancement in molecular genetic techniques, it is possible to assess the relationships between sperm functions and gene polymorphisms. In this study, variant calling analysis of RNA-Seq datasets was used to identify single nucleotide polymorphisms (SNPs) in boar spermatozoa and to explore the associations between SNPs and post-thaw semen quality. Assessment of post-thaw sperm quality characteristics showed that 21 boars were considered as having good semen freezability (GSF), while 19 boars were classified as having poor semen freezability (PSF). Variant calling demonstrated that most of the polymorphisms (67%) detected in boar spermatozoa were at the 3’-untranslated regions (3’-UTRs). Analysis of SNP abundance in various functional gene categories showed that gene ontology (GO) terms were related to response to stress, motility, metabolism, reproduction, and embryo development. Genomic DNA was isolated from sperm samples of 40 boars. Forty SNPs were selected and genotyped, and several SNPs were significantly associated with motility and membrane integrity of frozen-thawed (FT) spermatozoa. Polymorphism in SCLT1 gene was associated with significantly higher motility and plasma membrane integrity of FT spermatozoa from boars of the GSF group compared with those of the PSF group. Likewise, polymorphisms in MAP3K20, MS4A2, and ROBO1 genes were significantly associated with reduced cryo-induced lipid peroxidation and DNA damage of FT spermatozoa from boars of the GSF group. Candidate genes with significant SNP associations, including APPL1, PLBD1, FBXO16, EML5, RAB3C, OXSR1, PRICKLE1, and MAP3K20 genes, represent potential markers for post-thaw semen quality, and they might be relevant for future improvement in the selection procedure of boars for cryopreservation. The findings of this study provide evidence indicating that polymorphisms in genes expressed in spermatozoa could be considered as factors associated with post-thaw semen quality.


2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 112-113
Author(s):  
André F de Andrade

Abstract Identifying doses of post-thawed semen with high and low fertility potential is the main objective pursued by professionals and companies involved in commercializing frozen semen. In post-thaw semen, we can evaluate the motility characteristics and the integrity of the spermatozoa membranes and their DNA so that we can identify the number of cells that have the minimum characteristics to be considered with the potential to fertilize the oocyte. However, even with these analyzes using fluorescent probes, flow cytometry, and a computerized semen analysis system, it is impossible to predict the dose’s fertilizing potential accurately. The spermatozoa and seminal plasma originate from an individual (e.g., boar, bull, stallion), and even the spermatozoa and seminal plasma from this individual are different between different ejaculates. Factors such as genetics, age, ambiance, nutrition, and semen manipulation can alter the cryotolerance capacity of a given ejaculate, thus affecting its fertility potential. However, recent studies with assessments of proteomics, lipidomics, metabolomics, and miRNAs have associated cryotolerance and semen fertility with markers that can be evaluated before the ejaculate cryopreservation process—in this way, creating the possibility of selecting high fertility doses before the freezing process. Suppose these biological markers will conclusively make it possible to inform whether post-thawed semen dose has high or low fertility potential. In that case, only future research work verifying fertility will allow us to know. In the meantime, it is highly recommended to evaluate the post-thaw semen by assessing characteristics of motility, the integrity of the plasma and acrosomal membranes, and the integrity of DNA. Thus, ensuring that inseminations are carried out with the minimum number of sperm able to provide a high potential for fertility. These minimum numbers are related to the species and type of cryopreserved semen: conventional or sexed.


Animals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 283
Author(s):  
Marta F. Riesco ◽  
Mercedes Alvarez ◽  
Luis Anel-Lopez ◽  
Marta Neila-Montero ◽  
Cristina Palacin-Martinez ◽  
...  

The optimization of sperm cryopreservation protocols in ram is a feasible tool to reinforce artificial insemination technologies considering the desirable application of sperm by vaginal/cervical or transcervical deposition. Cryopreservation provokes different types of damage on spermatozoa and many of these detrimental effects are triggered by redox deregulation. For this reason, the antioxidant supplementation in sperm cryopreservation protocols to decrease reactive oxygen species (ROS) levels and to equilibrate redox status has been widely employed in different species. Despite this, more fertility trials are necessary to provide the definitive tool to ensure the antioxidant effectiveness on sperm quality. For this reason, in this work, we performed a multiparametric analysis of some previously tested antioxidants (crocin, GSH and Trolox) on ram sperm cryopreservation from field trials to sperm quality analyses focused on new strategies to measure redox balance. Attending to fertility trial, Trolox supplementation registered an improvement concerning to fertility (when we considered high fertility males) and multiple lambing frequency and other complementary and descriptive data related to lambing performance such as prolificacy and fecundity. This positive effect was more evident in multiple lambing frequency when we considered low fertility males than in global male analysis. In vitro analyses of sperm quality confirmed in vivo trials registering a positive effect on sperm viability and redox balance. In this study, we provided the definitive evidence that the role of trolox on redox balance maintenance has a direct effect on fertility parameters, such as prolificacy. The effectiveness of antioxidant treatments was tested, for the first time in ovine species, using an integrative and multiparametric approach combining in vivo and in vitro analyses and novel approaches, such as RedoxSYS. These types of strategies should be applied to improve sperm conservation methods and optimize AI technologies upgrading the correlation between in vitro and in vivo analyses.


2019 ◽  
Vol 102 (3) ◽  
pp. 730-739 ◽  
Author(s):  
Katrin Gegenfurtner ◽  
Thomas Fröhlich ◽  
Florian Flenkenthaler ◽  
Miwako Kösters ◽  
Sébastien Fritz ◽  
...  

Abstract Over the last decades, fertility of dairy cows has declined due to selection strategies focusing on milk yield. To study the effect of genetic merit for fertility on the proteome of the bovine uterine luminal fluid, Holstein heifers with low- and two groups of heifers with high-fertility index (high-fertility Holstein and Montbéliarde) were investigated. To focus on the maternal effect, heifers from all groups were synchronized and received on Day 7 high-quality embryos. Uterine luminal fluid from Day 19 pregnant heifers was analyzed in a holistic proteomic approach using nano-LC-MS/MS analysis combined with a label-free quantification approach. In total, 1737 proteins were identified, of which 597 differed significantly in abundance between the three groups. The vast majority of proteome differences was found comparing both high-fertility groups to the low-fertility Holstein group, showing that the genetic predisposition for fertility is prevalent regarding the uterine luminal fluid proteome. Evaluation of this dataset using bioinformatic tools revealed an assignment of higher abundant proteins in low-fertility Holstein to several metabolic processes, such as vitamin metabolic process, which comprises folate receptor alpha (FOLR1) and retinol-binding protein, indicating an involvement of disturbed metabolic processes in decreased fertility. Moreover, immune system-related proteins — lactotransferrin and chromogranin A — were enriched in low-fertility cows together with interferon tau 3 h and interferon tau-2. Our results indicate that the genetic merit for fertility leads to substantial quantitative differences at the level of proteins in uterine fluid of pregnant animals, thus altering the microenvironment for the early conceptus.


Author(s):  
J.S. Clark

Agroforests and woodlots offer Northland hill country farmers investment and diversification opportunities. Agroforests have less effect on the "whole farm" financial position than woodlots, especially where a progressive planting regime is adopted and where no further borrowing is required. Establishment and tending costs for agro-forests are lower, and returns come much sooner. The proven opportunity for continued grazing under trees established in this manner, apart from a short post-planting period, further enhances the agroforesty option. Even where there is reluctance on a farmer's part to plant trees on high fertility land, the expected financial returns from agroforests on low and medium fertility land will increase the overall long-term profitability and flexibility of the whole farming operation. Woodlots may be more appropriate on low fertility areas where weed reversion is likely. Joint ventures may be worth considering where farm finances are a limited factor. Keywords: On-farm forestry development, Northland hill country, agroforestry, woodlots, diversification, joint ventures, progressive planting regimes, grazing availability.


2020 ◽  
Vol 20 (2) ◽  
pp. 198-202 ◽  
Author(s):  
Mohammad Motamedifar ◽  
Yalda Malekzadegan ◽  
Parisa Namdari ◽  
Behzad Dehghani ◽  
Bahia Namavar Jahromi ◽  
...  

Introduction: Infertility considered as a social and public health issue and estimated that most of these infertile couples are residents of developing countries. Infectious diseases including the history of Sexually Transmitted Infections (STIs) may impact on male reproductive function. Therefore, the present study aimed to investigate the prevalence of bacterial contaminants of semen and probable association with sperm quality of infertile men in Iranian population. Methods: The study population consisted of 200 infertile men and 150 fertile men attending an infertility Center in southwestern Iran during the study period in 2015. The assessment of sperm parameters was according to the World Health Organization (WHO) guidelines. The presumptive pathogens were identified using standard microbiology tests and confirmed by specific PCR primers. Results: The prevalence of bacteriospermia in the semen of the infertile group was significantly higher than that in the fertile group (48% vs. 26.7%, P <0.001). The microbiological analysis of samples showed that the most abundant species of bacteria in semen of infertile men were Chlamydia trachomatis (12.5%) followed by Neisseria gonorrhoeae (11%). On the other hand, in the control group, Lactobacillus spp. (17.3%) was the most isolated pathogen. Results showed that the presence of N. gonorrhoeae, C. trachomatis, Mycoplasma genitalium, Haemophilus, and Klebsiella was significantly associated with sperm abnormality. Conclusion: Based on our findings, it seems that bacteriospermia is associated with alterations in the properties of semen which may lead to a decrease in the fertilization potential of sperm. Therefore, immediate and appropriate treatment is necessary before investigating every other possible cause of infertility.


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