boar sperm
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Author(s):  
Pascal D Kroh ◽  
Beate C Braun ◽  
Fan Lui ◽  
Peter Müller ◽  
Karin Müller

Abstract As a major spermadhesin first found in the seminal plasma of boars, AWN is described to fulfil a variety of reproduction related tasks. Although being the best investigated boar spermadhesin, information about its interaction with membranes is inconsistent. In this regard, previous reports locate AWN either inside or on the surface of sperm cells and at different regions, depending on the method and antibody used. Here, we localize native AWN in/on epididymal, ejaculated, capacitated and acrosome-reacted boar sperm using epifluorescence and electron microscopy, as well as an analysis of potential lipid binding partners of native and recombinant AWN. By applying a custom-made anti-AWN antibody, localization of AWN in the equatorial segment of ejaculated, capacitated and acrosome-reacted boar sperm was discovered. Electron microscopy showed that AWN is localized both on the sperm surface and on the cytoplasmic side of the plasma membrane, and in close vicinity to the nuclear and both acrosomal membranes of sperm. Analysis of epididymal sperm indicated migration of AWN from the retral postacrosomal part to the equatorial segment during the epididymal passage. In contrast to hypotheses claiming a specific association of AWN to phosphatidylethanolamine and in line with our previous study describing an interaction with phosphatidic acid, the current results show a rather electrostatically-driven binding mechanism of AWN to negative lipids. In conclusion, this work provides new insights into the arrangement of AWN in the equatorial segment that suggest a possible role in sperm-oocyte fusion.


2022 ◽  
Vol 34 (2) ◽  
pp. 280
Author(s):  
M. R. Ledwaba ◽  
M. L. Mphaphathi ◽  
M. A. Thema ◽  
C. M. Pilane ◽  
T. L. Nedambale

Cryobiology ◽  
2021 ◽  
Vol 103 ◽  
pp. 194-195
Author(s):  
Jordi Ribas-Maynou ◽  
Marc Llavanera ◽  
Yentel Mateo-Otero ◽  
Estela Garcia-Bonavila ◽  
Ariadna Delgado-Bermúdez ◽  
...  

Cryobiology ◽  
2021 ◽  
Vol 103 ◽  
pp. 190
Author(s):  
Norma A. Ramirez-Campos ◽  
Alicia Alcantar-Rodriguez ◽  
Oscar Gutierrez-Perez ◽  
Alfredo Medrano

Author(s):  
Mariana A Torres ◽  
Ana Carolina Pedrosa ◽  
Francisco José Novais ◽  
Diego V Alkmin ◽  
Bruce R Cooper ◽  
...  

Abstract Holding at room temperature is the first step in most boar semen cryopreservation protocols. It is well accepted that a holding time (HT) of 24 h increases sperm cryotolerance. However, the effect of HT on ejaculates with different freezability is not entirely clear. The aim of this study was to understand how HT influences spermatic and seminal plasma metabolite profiles of boar ejaculates and how these possible changes affect freezability. Twenty-seven ejaculates were collected and extended to 1:1 (v: v) with BTS and split into two aliquots. The first aliquot was cryopreserved without holding time (0 h), and the second was held at 17°C for 24 h before cryopreservation. Spermatozoa and seminal plasma were collected by centrifugation at two times, before HT (0 h) and after HT (24 h), and subsequently frozen until metabolite extraction and UPLC–MS analysis. After thawing, the semen samples were evaluated for kinetics, membrane integrity, mitochondrial potential, membrane lipid peroxidation, and fluidity. The ejaculates were then allocated into two phenotypes (good ejaculate freezers [GEF] and poor ejaculate freezers [PEF]) based on the percent reduction in sperm quality (%RSQ) as determined by the difference in total motility and membrane integrity between raw and post-thaw samples cryopreserved after 24 h of HT. The metabolic profile of the seminal plasma did not seem to influence ejaculate freezability, but that of the spermatozoa were markedly different between GEF and PEF. We identified a number of metabolic markers in the sperm cells (including inosine, hypoxanthine, creatine, ADP, niacinamide, spermine, and 2-methylbutyrylcarnitine) that were directly related to the improvement of ejaculate freezability during HT; these were components of metabolic pathways associated with energy production. Furthermore, PEF showed an up-regulation in the arginine and proline as well as the glutathione metabolism pathways. These findings help to better understand the effect of holding time on boar sperm freezability and propose prospective metabolic markers that may predict freezability; this has implications in both basic and applied sciences.


Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1360
Author(s):  
Janne Salin ◽  
Pasi Ohtonen ◽  
Maria A. Andersson ◽  
Hannu Syrjälä

Background: The causes and pathophysiological mechanisms of building-related symptoms (BRS) remain open. Objective: We aimed to investigate the association between teachers’ individual work-related symptoms and intrinsic in vitro toxicity in classrooms. This is a further analysis of a previously published dataset. Methods: Teachers from 15 Finnish schools in Helsinki responded to the symptom survey. The boar sperm motility inhibition assay, a sensitive indicator of mitochondrial dysfunction, was used to measure the toxicity of wiped dust and cultured microbial fallout samples collected from the teachers’ classrooms. Results: 231 teachers whose classroom toxicity data had been collected responded to the questionnaire. Logistic regression analysis adjusted for age, gender, smoking, and atopy showed that classroom dust intrinsic toxicity was statistically significantly associated with the following 12 symptoms reported by teachers (adjusted ORs in parentheses): nose stuffiness (4.1), runny nose (6.9), hoarseness (6.4), globus sensation (9.0), throat mucus (7.6), throat itching (4.4), shortness of breath (12.2), dry cough (4.7), wet eyes (12.7), hypersensitivity to sound (7.9), difficulty falling asleep (7.6), and increased need for sleep (7.7). Toxicity of cultured microbes was found to be associated with nine symptoms (adjusted ORs in parentheses): headache (2.3), nose stuffiness (2.2), nose dryness (2.2), mouth dryness (2.8), hoarseness (2.2), sore throat (2.8), throat mucus (2.3), eye discharge (10.2), and increased need for sleep (3.5). Conclusions: The toxicity of classroom dust and airborne microbes in boar sperm motility inhibition assay significantly increased teachers’ risk of work-related respiratory and ocular symptoms. Potential pathophysiological mechanisms of BRS are discussed.


BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Ziyue Qin ◽  
Wencan Wang ◽  
Malik Ahsan Ali ◽  
Yihan Wang ◽  
Yan Zhang ◽  
...  

Abstract Background Cryopreservation induces transcriptomic and epigenetic modifications that strongly impairs sperm quality and function, and thus decrease reproductive performance. N6-methyladenosine (m6A) RNA methylation varies in response to stress and has been implicated in multiple important biological processes, including post-transcriptional fate of mRNA, metabolism, and apoptosis. This study aimed to explore whether cryopreservation induces m6A modification of mRNAs associated with sperm energy metabolism, cryoinjuries, and freezability. Results The mRNA and protein expression of m6A modification enzymes were significantly dysregulated in sperm after cryopreservation. Furthermore, m6A peaks were mainly enriched in coding regions and near stop codons with classical RRACH motifs. The mRNAs containing highly methylated m6A peaks (fts vs. fs) were significantly associated with metabolism and gene expression, while the genes with less methylated m6A peaks were primarily involved in processes regulating RNA metabolism and transcription. Furthermore, the joint analysis of DMMGs and differentially expressed genes indicated that both of these play a vital role in sperm energy metabolism and apoptosis. Conclusions Our study is the first to reveal the dynamic m6A modification of mRNAs in boar sperm during cryopreservation. These epigenetic modifications may affect mRNA expression and are closely related to sperm motility, apoptosis, and metabolism, which will provide novel insights into understanding of the cryoinjuries or freezability of boar sperm during cryopreservation.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
S Galal ◽  
C Jones ◽  
K Coward

Abstract Study question Do solid silica nanoparticles qualify as a new research tool for the in vitro transfer of compounds into gametes prior to Assisted Reproductive Technology (ART). Summary answer Solid silica nanoparticles (SSNPs) could be used as an intra-gamete delivery system to deliver therapeutic biomolecules into gametes prior to ART. What is known already Sperm-mediated gene transfer (SMGT) results in the production of transgenic embryos; however, the success rate of this technique is low. Nanoparticles are an efficient intra-cellular delivery system in vitro. Naturally cell-secreted nanoparticles are involved in the development of gametes. Mesoporous silica nanoparticles have been shown to carry large amounts of compounds and to interact with gametes without toxic effects, thus providing an alternative to naturally secreted nanoparticles. However, this technique is associated with some limitations, such as the size of these nanoparticles. SSNPs can be synthesised on a smaller nanoscale, thus providing higher potential to penetrate gametes and delivering biomolecules. Study design, size, duration This was an experimental in vitro study that investigated the effects of SSNPs on the motility of boar sperm and the degeneration of hamster oocytes, as determined by ooplasm shrinkage. Participants/materials, setting, methods SSNPs (20 nm) were conjugated with fluorescein diacetate–5-maleimide (FDA5M), a fluorescent protein. FDA5M-labelled SSNPS were incubated with boar sperm (N = 3) at 10 and 30µg/ml/107 sperm for four-hours. Motility parameters were assessed by computer-assisted sperm analysis (CASA). Binding potential was evaluated by fluorescent microscopy. Hamster oocytes (7 oocytes/group) were incubated with FDA5M-labelled SSNPs at 100, 150, and 300µg/ml, for two-hours; ooplasm shrinkage was evaluated. Time/matched control sperm was incubated in phosphate-buffered saline and oocytes in KSOM. Main results and the role of chance Exposure to FDA5M-labelled SSNPs did not affect total or progressive sperm motility (P = 0.6735 and 0.9606, respectively), average-path velocity or straight-line velocity after 4-hours of incubation (P = 0.7459 and 0.8696, respectively) compared to controls. SSNPs at 10 µg/ml significantly increased sperm curvilinear velocity after 1-hour (P = 0.0495) and linearity and straightness after 4-hours (P = 0.0389 and 0.0312, respectively). SSNPs at 30 µg/ml significantly increased sperm linearity after 3- and 4-hours (P = 0.0384 and 0.005, respectively). The proportion of sperm showing green fluorescence was significantly higher in the 30µg/ml dose of SSNPs than the 10µg/ml dose after 4-hours (P < 0.00001). In oocytes, the zona pellucida remained morphologically intact and the ooplasm exhibited green fluorescence. The ooplasm of 42% of the oocytes at 300µg/ml showed ooplasm shrinkage (a sign of degeneration); no oocytes showed shrinkage at doses of 100 and 150µg/ml of SSNPs. The green fluorescence in the sperm head and the ooplasm indicated the ability of SSNPs to spontaneously interact non-invasively with these gametes either by surface association or by cell-internalisation. This could provide a safe and non-invasive intra-gamete delivery system for research purposes and clinical therapy. This system could be used to deliver specific agents into gametes prior to ART to improve outcomes. Limitations, reasons for caution The SSNPs are non-biodegradable; it remains unknown as to how gametes or embryos might react with SSNPs over long time periods. The nanotoxicity of SSNPs has not yet been investigated over the long term. SSNPs have still to be tested with embryos to evaluate their effect on embryonic development. Wider implications of the findings: SSNPs could be functionalised to target the nucleus of mammalian gametes and embryos to act as a carrier for oligonucleotides and genes to correct chromosomal abnormalities and to provide genetic therapy in these gametes and embryos to treat hereditary diseases before intra-uterine transfer. Trial registration number Not applicable


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