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Veterinary World Apr 2024The quality of canine sperm can be influenced by many factors, such as breed, body weight, age, ejaculatory frequency, nutrition, and environment. In the UK, it is...
BACKGROUND AND AIMS
The quality of canine sperm can be influenced by many factors, such as breed, body weight, age, ejaculatory frequency, nutrition, and environment. In the UK, it is common practice for standard Bull Terriers (SBT) and miniature Bull Terriers (MBT) to require male donors during a short breeding period. The aim of this study was to evaluate the effect of semen collection frequency on ejaculate volume and nine sperm parameters in SBT and MBT males, considering age and body condition score (BCS).
MATERIALS AND METHODS
Ejaculates from six adult SBTs and four MBTs were collected 5 times at two consecutive intervals (Time Series [TS]1, 24 h . TS2, 48 h), 1 week apart. Ejaculate volume, concentration, total output, viability (live sperm), subjective total motility, vigor, and total morphological defects, including head, midpiece, and tail defects of sperm, were evaluated. A multivariable mixed linear model for repeated measures was used to analyze the effects of semen collection frequency, age, breed, and BCS on ejaculate volume and sperm parameters.
RESULTS
Semen collection frequency, age, and, to a lesser extent, breed, and BCS significantly affected sperm parameters. Semen collection frequency affected all sperm parameters (p < 0.05) but not ejaculate volume (p > 0.05). Total sperm output, sperm vigor, total motility, and tail defects decreased (p < 0.05) at the end of TS1. However, sperm parameters remained relatively constant (p > 0.05) in TS2 between semen collection sessions. Overall, poorer sperm parameters were observed in older dogs (aged 5-8 years) than in younger dogs (aged 4 years). MBT produced less (p < 0.001) ejaculate volume (3.2 ± 0.2 mL . 4.3 ± 0.2 mL: Least Squares Mean ± Standard Error of Mean), lower total sperm output (221.8 ± 19.2 × 10 vs. 348.6 ± 19.2 × 10) and lower total morphological defects (25.0 ± 1.1% . 31.3 ± 0.9%), and a higher percentage of live sperm (77.0 ± 1.4% . 71.7 ± 1.1%) than SBT. In addition, a BCS of 4 positively influenced (p < 0.05) viability, vigor, and total sperm motility.
CONCLUSION
Despite differences in age, breed, and BCS, better sperm parameter values were observed in all semen collection sessions. However, intensive semen collection (TS1) appears to be less effective in maintaining good sperm quality. For breeding or artificial insemination purposes, a 48-h interval between collection sessions is recommended for both breeds. The results of this study could be used to further optimize assisted reproductive technologies in both breeds.
PubMed: 38798297
DOI: 10.14202/vetworld.2024.820-828 -
The World Journal of Men's Health Apr 2024Infertility is a global health issue and nutrition plays a significant role in fertility outcomes. We aimed to investigate the cross-sectional and prospective...
PURPOSE
Infertility is a global health issue and nutrition plays a significant role in fertility outcomes. We aimed to investigate the cross-sectional and prospective associations of glycemic index (GI) and glycemic load (GL) with semen quality parameters in a cohort of healthy young men.
MATERIALS AND METHODS
The study included 106 men aged 18-35 years from the FERTINUTS trial. Dietary intake was estimated through 3-day dietary records and several semen parameters were assessed. Multivariable linear regression analysis with the Least Absolute Shrinkage and Selection Operator (LASSO) approach was employed.
RESULTS
The cross-sectional analysis revealed positive associations between GI and GL and total sperm count, sperm concentration, and total motility. In the prospective analysis, baseline GI was associated with increases in pH, vitality, immotile sperm or abnormal midpiece and decreases in total sperm count and motility. Conversely, GL was positively associated with changes in vitality and total sperm count.
CONCLUSIONS
While these findings suggest that GI may have adverse effects on several sperm quality parameters, the results were not consistently observed in the cross-sectional analysis. However, GL was consistently associated with better sperm quality in both analyses. The impact of carbohydrate quality and quantity on fertility remains uncertain and larger prospective studies are needed.
PubMed: 38772538
DOI: 10.5534/wjmh.230328 -
Animal Reproduction 2024This study aimed to analyze the characteristics of the HSP70 gene and protein in spermatozoa of Bali bulls of different age groups and to examine its potential as a...
This study aimed to analyze the characteristics of the HSP70 gene and protein in spermatozoa of Bali bulls of different age groups and to examine its potential as a biomarker determining bull fertility. This study used frozen semen produced from six Bali bulls divided into two groups based on age (≤ 9 years and ≥ 12 years). Parameters of frozen semen quality analyzed included sperm motility and kinetics using computer-assisted semen analysis, sperm morphological defects using Diff-Quick staining, acrosome integrity using FITC-PNA staining, and DNA fragmentation using acridine orange staining. HSP70 gene expression characterization was analyzed using qRT-PCR, and HSP70 protein abundance was analyzed using enzyme immunoassays. Fertility field data were obtained by analyzing the percentage conception rate for each bull based on the artificial insemination service data contained in the Indonesian-integrated system of the National Animal Health Information System (iSIKHNAS). The results showed significant differences (P<0.05) in total and progressive motility, morphological defects of the neck and midpiece, and tail of sperm, and acrosome integrity between the age groups of Bali bulls. HSP70 gene expression and protein abundance showed no significant differences (P>0.05) in different age groups. HSP70 gene expression correlated with fertility rate (P<0.05). Age affected several semen quality parameters but did not affect HSP70 gene expression and protein abundance. The HSP70 gene molecule could be a biomarker that determines the fertility of Bali bulls.
PubMed: 38756622
DOI: 10.1590/1984-3143-AR2023-0048 -
Cellular & Molecular Biology Letters May 2024By analyzing a mouse Interspecific Recombinant Congenic Strain (IRCS), we previously identified a quantitative trait locus (QTL), called Mafq1 on mouse chromosome 1,...
By analyzing a mouse Interspecific Recombinant Congenic Strain (IRCS), we previously identified a quantitative trait locus (QTL), called Mafq1 on mouse chromosome 1, that is associated with male hypofertility and ultrastructural sperm abnormalities. Within this locus, we identified a new candidate gene that could be implicated in a reproductive phenotype: Tex44 (Testis-expressed protein 44). We thus performed a CRISPR/Cas9-mediated complete deletion of this gene in mice in order to study its function. Tex44-KO males were severely hypofertile in vivo and in vitro due to a drastic reduction of sperm motility which itself resulted from important morphological sperm abnormalities. Namely, Tex44-KO sperm showed a disorganized junction between the midpiece and the principal piece of the flagellum, leading to a 180° flagellar bending in this region. In addition, the loss of some axonemal microtubule doublets and outer dense fibers in the flagellum's principal piece has been observed. Our results suggest that, in mice, TEX44 is implicated in the correct set-up of the sperm flagellum during spermiogenesis and its absence leads to flagellar abnormalities and consequently to severe male hypofertility.
Topics: Animals; Male; Infertility, Male; Sperm Motility; Sperm Tail; Mice; Mice, Knockout; Spermatozoa; Spermatogenesis; Flagella; Mice, Inbred C57BL; CRISPR-Cas Systems
PubMed: 38750428
DOI: 10.1186/s11658-024-00587-5 -
Theriogenology Jul 2024The freeze-thawing process induces osmotic changes that may affect the membrane domain location of aquaporins' (AQP) in spermatozoa. Recent studies suggest that changes...
The freeze-thawing process induces osmotic changes that may affect the membrane domain location of aquaporins' (AQP) in spermatozoa. Recent studies suggest that changes in AQP3 localization allows better sperm osmo-adaptation, improving the cryoresistance. Ultra-rapid freezing is an alternative cryopreservation technique that requires less equipment than conventional freezing, and it is faster, simpler and can be used in the field. This study aimed to determine the influence of freezing-thawing rates (slow (control) vs. ultra-rapid) on AQP3 expression and location in the spermatozoa from small ruminants (sheep and goats) and its relationship with sperm cryo-damage. Spermatozoa were collected from 10 Merino rams and 10 Murciano-Granadina bucks. The presence and distribution of AQP3 were assessed by Western blotting and immunocytochemistry (ICC), employing a commercial rabbit polyclonal antibody. Sperm motility was CASA system-analyzed, and membrane and acrosome integrity assessed by fluorescence (PI/PNA-FITC). Western blotting did not detect a significant effect of freezing-thawing rate on the amount of AQP3 while ICC found freezing-thawing rate affecting AQP3 location (P < 0.05). In both species, the percentages of spermatozoa showing AQP3 in the post-acrosome region, mid-piece, and principal piece of the tail were greater in samples cryopreserved by slow freezing-thawing (control) than ultra-rapid freezing-thawing rates (P < 0.05). Spermatozoa cryopreserved using ultra-rapid freezing-thawing showed decrease motility, plasma membrane, and acrosome integrity (P < 0.05), which might be related, at least in part, to a lower expression of AQP3. In conclusion, the cooling rate modifies the location of AQP3 in spermatozoa of sheep and goat, which might be associated with sperm cryosurvival.
Topics: Animals; Male; Goats; Aquaporin 3; Spermatozoa; Cryopreservation; Sheep; Semen Preservation; Freezing; Sperm Motility
PubMed: 38663138
DOI: 10.1016/j.theriogenology.2024.04.008 -
Human Reproduction Open 2024Is the mutation causative for male infertility?
STUDY QUESTION
Is the mutation causative for male infertility?
SUMMARY ANSWER
Our collected data underline the complex and devastating effect of the single-gene mutation on the testicular molecular network, leading to male reproductive failure.
WHAT IS KNOWN ALREADY
Recent data have revealed mutations in genes related to axonemal dynein arms as causative for morphology and motility abnormalities in spermatozoa of infertile males, including dysplasia of fibrous sheath (DFS) and multiple morphological abnormalities in the sperm flagella (MMAF). The nexin-dynein regulatory complex (N-DRC) coordinates the dynein arm activity and is built from the DRC1-DRC7 proteins. DRC5 (TCTE1), one of the N-DRC elements, has already been reported as a candidate for abnormal sperm flagella beating; however, only in a restricted manner with no clear explanation of respective observations.
STUDY DESIGN SIZE DURATION
Using the CRISPR/Cas9 genome editing technique, a mouse gene knockout line was created on the basis of the C57Bl/6J strain. The mouse reproductive potential, semen characteristics, testicular gene expression levels, sperm ATP, and testis apoptosis level measurements were then assessed, followed by visualization of N-DRC proteins in sperm, and protein modeling . Also, a pilot genomic sequencing study of samples from human infertile males (n = 248) was applied for screening of variants.
PARTICIPANTS/MATERIALS SETTING METHODS
To check the reproductive potential of KO mice, adult animals were crossed for delivery of three litters per caged pair, but for no longer than for 6 months, in various combinations of zygosity. All experiments were performed for wild-type (WT, control group), heterozygous and homozygous male mice. Gross anatomy was performed on testis and epididymis samples, followed by semen analysis. Sequencing of RNA (RNAseq; Illumina) was done for mice testis tissues. STRING interactions were checked for protein-protein interactions, based on changed expression levels of corresponding genes identified in the mouse testis RNAseq experiments. Immunofluorescence staining was performed to detect the N-DRC complex proteins: Tcte1 (Drc5), Drc7, Fbxl13 (Drc6), and Eps8l1 (Drc3) in mouse spermatozoa. To determine the amount of ATP in spermatozoa, the luminescence level was measured. In addition, immunofluorescence staining was performed to check the level of apoptosis via caspase 3 visualization on mouse testis samples. DNA from whole blood samples of infertile males (n = 137 with non-obstructive azoospermia or cryptozoospermia, n = 111 samples with a spectrum of oligoasthenoteratozoospermia, including n = 47 with asthenozoospermia) was extracted to perform genomic sequencing (WGS, WES, or Sanger). Protein prediction modeling of human-identified variants and the exon 3 structure deleted in the mouse knockout was also performed.
MAIN RESULTS AND THE ROLE OF CHANCE
No progeny at all was found for the homozygous males which were revealed to have oligoasthenoteratozoospermia, while heterozygous animals were fertile but manifested oligozoospermia, suggesting haploinsufficiency. RNA-sequencing of the testicular tissue showed the influence of mutations on the expression pattern of 21 genes responsible for mitochondrial ATP processing or linked with apoptosis or spermatogenesis. In males, the protein was revealed in only residual amounts in the sperm head nucleus and was not transported to the sperm flagella, as were other N-DRC components. Decreased ATP levels (2.4-fold lower) were found in the spermatozoa of homozygous mice, together with disturbed tail:midpiece ratios, leading to abnormal sperm tail beating. Casp3-positive signals (indicating apoptosis) were observed in spermatogonia only, at a similar level in all three mouse genotypes. Mutation screening of human infertile males revealed one novel and five ultra-rare heterogeneous variants (predicted as disease-causing) in 6.05% of the patients studied. Protein prediction modeling of identified variants revealed changes in the protein surface charge potential, leading to disruption in helix flexibility or its dynamics, thus suggesting disrupted interactions of TCTE1 with its binding partners located within the axoneme.
LARGE SCALE DATA
All data generated or analyzed during this study are included in this published article and its supplementary information files. RNAseq data are available in the GEO database (https://www.ncbi.nlm.nih.gov/geo/) under the accession number GSE207805. The results described in the publication are based on whole-genome or exome sequencing data which includes sensitive information in the form of patient-specific germline variants. Information regarding such variants must not be shared publicly following European Union legislation, therefore access to raw data that support the findings of this study are available from the corresponding author upon reasonable request.
LIMITATIONS REASONS FOR CAUTION
In the study, the fertilization performance of sperm from homozygous male mice was not checked.
WIDER IMPLICATIONS OF THE FINDINGS
This study contains novel and comprehensive data concerning the role of in male infertility. The gene is the next one that should be added to the 'male infertility list' because of its crucial role in spermatogenesis and proper sperm functioning.
STUDY FUNDING/COMPETING INTERESTS
This work was supported by National Science Centre in Poland, grants no.: 2015/17/B/NZ2/01157 and 2020/37/B/NZ5/00549 (to M.K.), 2017/26/D/NZ5/00789 (to A.M.), and HD096723, GM127569-03, NIH SAP #4100085736 PA DoH (to A.N.Y.). The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.
PubMed: 38650655
DOI: 10.1093/hropen/hoae020 -
Animals : An Open Access Journal From... Mar 2024Anthropogenic radiofrequency electromagnetic radiation (RF-EMR) from wireless technologies has increased dramatically. The boar semen used for artificial insemination is...
The Distribution of Boars Spermatozoa in Morphometrically Distinct Subpopulations after In Vitro Exposure to Radiofrequency Electromagnetic Radiation at 2500 MHz and Their Motility.
Anthropogenic radiofrequency electromagnetic radiation (RF-EMR) from wireless technologies has increased dramatically. The boar semen used for artificial insemination is essential in sustaining the pig industry, and additionally it is also exposed to the effects of the RF-EMR of wireless technologies. Furthermore, there are no data on the effects of RF-EMR on semen quality, and this is the first analysis of sperm's morphometric parameters for assessing the effect of RF-EMR on the spermatozoa subpopulations of boars. This study investigated the effect of RF-EMR on in vitro exposed breeding boar semen spermatozoa motility and the proportions of spermatozoa subpopulations according to their morphometric head and tail parameters. The semen samples of 12 boars were divided into control and experimental groups. The samples in the experimental group were exposed in a gigahertz transverse electromagnetic chamber at a frequency of 2500 MHz (the frequency band used in 5G technology) and an electric field strength of 10 Vm for two hours. After exposure, the spermatozoa motility was evaluated for both groups. A morphometric analysis of the semen smears was performed using SFORM software (Version 1.0; VAMS, Zagreb, Croatia). The progressive spermatozoa motility was significantly reduced in the experimental group (74.7% vs. 85.7%). PC analysis and cluster analysis revealed two spermatozoa subpopulations: S1, spermatozoa with a more regular head shape and a smaller midpiece outline, and S2, spermatozoa with a more elongated head shape and a larger midpiece outline. The experimental semen samples had a greater proportion of the S1 spermatozoa subpopulation (68.2% vs. 64.4%). The effect of RF-EMR at 2500 MHz on the in vitro exposed boar semen resulted in decreased progressive spermatozoa motility and a lower proportion of the spermatozoa subpopulation with a higher fertilizing potential.
PubMed: 38539926
DOI: 10.3390/ani14060828 -
Journal of Animal Science Jan 2024Fertility is economically important but is hard to quantify and measure in breeding programs which has led extensive breeding programs to ignore fertility in their...
Fertility is economically important but is hard to quantify and measure in breeding programs which has led extensive breeding programs to ignore fertility in their selection criteria. While female fertility traits have been extensively researched, male fertility traits have been largely ignored. It is estimated that 20% to 40% of bulls have sub-fertility, reducing the number of calves born and profits, highlighting the importance of investigating bull fertility. The most practical measure of male fertility is a bull breeding soundness evaluation (BBSE) which assesses structure as well as semen quality and quantity. Generally, traits recorded in a BBSE are neither genetically evaluated nor used for selection in breeding programs. All traits recorded during a BBSE were analyzed through a series of univariate and bivariate linear mixed models using a genomic relationship matrix to estimate genetic parameters. All genotype and phenotype data were obtained from a tropical composite commercial cattle population and imputed to 27,638 single-nucleotide polymorphisms (SNPs) with a total of 2,613 genotyped animals with BBSE records ranging from 616 to 826 animals depending on the trait. The heritabilities of the 27 traits recorded during a BBSE ranged from 0.02 to 0.49. Seven of the male fertility traits were recommended to be included in a breeding program based on their heritability and their phenotypic and genetic correlations. These traits are scrotal circumference, percent normal sperm, proximal droplets, distal midpiece reflex, knobbed acrosomes, vacuoles/teratoids, and sheath score. Using these seven traits in a breeding program would result in higher calving rates, increasing production and profitability.
Topics: Cattle; Male; Animals; Female; Semen Analysis; Semen; Phenotype; Fertility; Reproduction
PubMed: 38477357
DOI: 10.1093/jas/skae069 -
Journal of Translational Medicine Mar 2024Mitochondria produce adenosine triphosphate through respiratory activities to power sperm differentiation and motility, and decreased mitochondrial respiratory activity...
BACKGROUND
Mitochondria produce adenosine triphosphate through respiratory activities to power sperm differentiation and motility, and decreased mitochondrial respiratory activity can result in poor sperm motility and asthenospermia. The mitochondrial sheath is a component of the mid-piece of the sperm flagellum, and dysfunction of the sheath can reduce sperm motility and cause male infertility. The membrane occupation and recognition nexus-motif protein 2 (MORN2) is testis enriched in mice, and the MORN motif was reported to play a role in the regulation of bioelectrical signal homeostasis in cardiomyocytes.
METHODS
We generated Morn2 mice using CRISPR/Cas9 and evaluated the potential functions of MORN2 in spermiogenesis through histological analysis, fertility examination, RT-PCR, CASA, immunofluorescence, TUNEL, electron microscopy analysis, mitochondrial energy metabolism analysis, etc. RESULTS: The Morn2 mice were infertile, and their sperm showed severe motility defects. Morn2 sperm also had abnormal morphology characterized by bent heads, aberrant mitochondrial sheath formation, lower mitochondrial membrane potential, higher levels of reactive oxygen species, and decreased mitochondrial respiratory activity.
CONCLUSIONS
Our study demonstrates that MORN2 is essential for male fertility and indicates that MORN2 functions in mitochondrial sheath formation and regulates mitochondrial respiratory activity.
Topics: Animals; Male; Mice; Energy Metabolism; Fertility; Mitochondria; Semen; Sperm Motility
PubMed: 38443933
DOI: 10.1186/s12967-024-05010-3 -
Cureus Jan 2024Infertility, defined as the inability to conceive after 12 months of unprotected sexual activity, affects millions globally. Approximately 80% of cases have identifiable...
Infertility, defined as the inability to conceive after 12 months of unprotected sexual activity, affects millions globally. Approximately 80% of cases have identifiable causes, including endometriosis, tubal obstruction, ovulatory dysfunction, and male sperm abnormalities. Lifestyle factors, such as smoking and obesity, also impact fertility. Sperm morphology, a key factor in male infertility, often presents as teratozoospermia, with defects in the head, midpiece, or tail. Poor ovarian reserve, indicated by low anti-mullerine hormone (AMH) and antra-follicular count (AFC) values, contributes to female infertility, often exacerbated by age-related factors. Elevated follicle-stimulating hormone (FSH) levels further diminish oocyte quantity and quality. Intracytoplasmic Sperm Injection (ICSI), a micromanipulation technique aiding infertile couples, may face challenges in detecting subtle sperm morphology defects. Advanced methods like Motile Sperm Organelle Morphological Examination (MSOME) and Intracytoplasmic Morphologically Selected Sperm Injection (IMSI) under high magnification enhance sperm selection accuracy. We present the case of a 36-year-old woman and her 42-year-old husband who sought assistance after seven years of infertility. Previous Intrauterine injection (IUI) and ICSI attempts failed due to the wife's low ovarian reserve and elevated FSH, compounded by the husband's teratozoospermia. Their earlier In-Vitro Fertilization (IVF) experience yielded a single poor-quality oocyte, hindering blastocyst formation. Investigations revealed the wife's poor AFC, AMH of 0.033ng/ml, and FSH at 24IU/L. Her medical history included hypertension and gallbladder removal. The husband exhibited 98% defective sperm, devoid of a substance abuse history. The wife's family had a polycystic ovarian syndrome (PCOS) history, and her low AMH and AFC yielded only three poor-quality oocytes during the current assessment. Oocytes were retrieved, and sperm were selected with the help of IMSI. After ICSI, the patient successfully conceived.
PubMed: 38435865
DOI: 10.7759/cureus.53268