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2.
Reprod Domest Anim ; 59(10): e14728, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39350734

RESUMO

Combining cholesterol-loaded methyl-ß-cyclodextrin (CD-CHL) with vitamin E-loaded methyl-ß-cyclodextrin (CD-Vit E) to combat cold shock and oxidative stress during sperm cryopreservation in soybean lecithin extenders remains unexplored. Thus, the current study aimed to investigate the effect of treating bull sperm with CD-CHL and CD-Vit E prior to cryopreservation in a soybean lecithin extender. Sperm collected from eight fertile bulls were pooled and split into six aliquots. Five aliquots were treated, in a Tris-based extender, with CD-CHL (2 mg/120 × 106 cells/mL) and either 0, 0.5, 1.0, 1.5 or 2 mg CD-Vit E/120 × 106 cells/mL. The control aliquot was diluted in a Tris-based extender without further supplementation. After incubation at 22°C for 15 min and addition of a soybean lecithin extender, all aliquots were equilibrated for 2 h at 4°C and then cryopreserved in liquid nitrogen. Computer-assisted sperm analysis (CASA) was used to explore the different sperm motility parameters, hypo-osmotic swelling test to determine membrane functionality and fluorescein isothiocyanate-conjugated Aeachis hypogaea (peanut) agglutinin (FITC-PNA) to quantify acrosome integrity. The effect of oxidative stress on the sperm membrane was assessed through lipid peroxidation measurement. Compared to control, CD-CHL alone improved significantly (p < 0.05) all CASA motility parameters, membrane functionality and acrosome integrity of thawed sperm. The membrane functionality was more significantly (p < 0.05) improved when 0.5 mg CD-Vit E was combined with CD-CHL. Concerning lipid peroxidation, no significant differences (p > 0.05) in malondialdehyde (MDA) levels were registered between groups. In conclusion, the combination of CD-CHL and CD-Vit E demonstrated a significant positive effect on the cryopreservation of bull sperm in a soybean lecithin extender.


Assuntos
Colesterol , Criopreservação , Crioprotetores , Glycine max , Preservação do Sêmen , Motilidade dos Espermatozoides , Espermatozoides , Vitamina E , Masculino , Animais , Criopreservação/veterinária , Criopreservação/métodos , Bovinos , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Vitamina E/farmacologia , Crioprotetores/farmacologia , Colesterol/farmacologia , Espermatozoides/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Glycine max/química , Lecitinas/farmacologia , beta-Ciclodextrinas/farmacologia , Ciclodextrinas/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Acrossomo/efeitos dos fármacos
3.
Arch Ital Urol Androl ; 96(3): 12595, 2024 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-39356024

RESUMO

PURPOSE: To determine the effect of electromagnetic waves of mobile phone stations on several sperm parameters and the male reproductive system. METHODS: This observational study was performed on 216 subjects, aged 18-60 years. Two equal groups of subjects were assigned to group A (study group) if they were living close to cell phone tower stations for at least 6 months and group B (control group) formed from individuals living 100 meters away from cell phone tower stations. Every subject underwent a comprehensive history taking, a clinical assessment, and laboratory testing. RESULTS: Regarding morphology index in the studied groups, the exposed group exhibited a trend of reduced percentage of normal morphology compared to the non-exposed group, with no statistical difference between the two groups. Regarding the total sperm motility (A+B+C) and progressive sperm motility (A+B) in the studied groups, the exposed group showed a trend of decreased total sperm motility and of progressive sperm motility in contrast to the non-exposed group, with no statistical difference between the two groups. CONCLUSIONS: Personal wrong lifestyles with exposure to electromagnetic waves have shown a trend towards a reduced percentage of normal morphology and reduced motility although nonstatistically significant compared with non-exposed populations.


Assuntos
Telefone Celular , Ondas de Rádio , Motilidade dos Espermatozoides , Humanos , Masculino , Adulto , Adolescente , Adulto Jovem , Motilidade dos Espermatozoides/efeitos da radiação , Ondas de Rádio/efeitos adversos , Pessoa de Meia-Idade , Radiação Eletromagnética , Fertilidade/efeitos da radiação , Espermatozoides/efeitos da radiação , Infertilidade Masculina/etiologia , Exposição Ambiental/efeitos adversos
4.
Trop Anim Health Prod ; 56(8): 318, 2024 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-39356339

RESUMO

Seasonal variations significantly impact buffalo bull semen production and quality, particularly during the summer months. Understanding the genetic basis of these changes is important for managing bull fertility and improving sperm quality. The present study focused on characterizing and identifying polymorphisms in chromatin remodeling genes, protamines (PRMs) and Transition Nuclear Proteins (TNPs) in Murrah buffalo bulls with varying semen quality due to seasonal effects. Our findings revealed none of the coding region variation in PRM1, PRM2, TNP1, and TNP2, these genes are highly conserved in buffalo. Two intronic variants were identified, including G16C in PRM1 intron 1 and intronic SNP in PRM2 intron 1 (G96A). The complete CDS of consensus sequence of bubaline PRM1 was 86.3% identical and 94.1% similar to the bovine PRM1. Whereas the complete CDS of consensus sequence of bubaline TNP2 was 78.2% identical and 91.0% similar to bovine TNP2. Further, no statistically significant differences in the fold change of TNP1, TNP2, PRM1, and PRM2 levels between the hot summer SNA and SA groups and the winter SNA and SA groups This study represents the first comprehensive report on the characterization of bubaline PRM1 (complete CDS), PRM2 (partial CDS), TNP1 (partial CDS), and TNP2 (complete CDS) genes in buffalo sperm cells. Results of the study, clearly indicate that the genes associated with protamine (PRM1 and TNP2) are highly conserved in Bubalus bubalis. Understanding these genetic underpinnings can have implications for improving buffalo bull fertility and semen quality.


Assuntos
Búfalos , Montagem e Desmontagem da Cromatina , Espermatozoides , Animais , Búfalos/genética , Masculino , Espermatozoides/fisiologia , Protaminas/genética , Protaminas/metabolismo , Estações do Ano , Análise do Sêmen/veterinária , Polimorfismo Genético , Proteínas Cromossômicas não Histona
5.
Nat Commun ; 15(1): 8128, 2024 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-39362867

RESUMO

Pacific bluefin tuna (Thunnus orientalis) remains heavily depleted due to overexploitation. Aquaculture and stock enhancement based on artificial seedlings could be effective solutions to this problem. However, widespread adoption of seedling production is limited because spawning in captivity of bluefin tuna, a large pelagic top predator, requires much space, time, cost, and labour. To downsize and shorten their generation time, here, we show that xenogeneic germ cell transplantation enables a surrogate production of functional bluefin tuna sperm in hybrid little tuna (genus: Euthynnus), which attain sexual maturity at a smaller body size and shorter time. Intraperitonially transplanted bluefin tuna germ cells migrate towards and are incorporated into the gonads of little tuna larvae. These recipients produce functional donor-derived bluefin tuna sperm at a mere 8 months of age and 1 kg of body size. This result represents an advance toward compact, low-cost, and time-efficient seedling production that will improve the efficiency of bluefin tuna aquaculture.


Assuntos
Espermatozoides , Atum , Animais , Masculino , Atum/fisiologia , Espermatozoides/fisiologia , Aquicultura/métodos , Feminino , Células Germinativas , Tamanho Corporal , Maturidade Sexual/fisiologia
6.
Nat Commun ; 15(1): 8477, 2024 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-39353950

RESUMO

Seminal fluid is rich in sugars, but their role beyond supporting sperm motility is unknown. In this study, we found Drosophila melanogaster males transfer a substantial amount of a phospho-galactoside to females during mating, but only half as much when undernourished. This seminal substance, which we named venerose, induces an increase in germline stem cells (GSCs) and promotes sperm storage in females, especially undernourished ones. Venerose enters the hemolymph and directly activates nutrient-sensing Dh44+ neurons in the brain. Food deprivation directs the nutrient-sensing neurons to secrete more of the neuropeptide Dh44 in response to infused venerose. The secreted Dh44 then enhances the local niche signal, stimulating GSC proliferation. It also extends the retention of ejaculate by females, resulting in greater venerose absorption and increased sperm storage. In this study, we uncovered the role of a sugar-like seminal substance produced by males that coordinates reproductive responses to nutritional challenges in females.


Assuntos
Drosophila melanogaster , Reprodução , Comportamento Sexual Animal , Animais , Masculino , Feminino , Drosophila melanogaster/fisiologia , Drosophila melanogaster/metabolismo , Comportamento Sexual Animal/fisiologia , Reprodução/fisiologia , Espermatozoides/metabolismo , Espermatozoides/fisiologia , Sêmen/metabolismo , Sêmen/química , Proteínas de Drosophila/metabolismo , Neurônios/metabolismo , Neurônios/fisiologia , Açúcares/metabolismo , Neuropeptídeos/metabolismo , Estresse Fisiológico , Hemolinfa/metabolismo , Encéfalo/metabolismo , Motilidade dos Espermatozoides/fisiologia
7.
BMC Biol ; 22(1): 207, 2024 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-39278917

RESUMO

BACKGROUND: The importance of parental diet in relation to eventual offspring health is increasing in prominence due to the increased frequency of parents of reproductive age consuming poor diets. Whilst maternal health and offspring outcome have been studied in some detail, the paternal impacts are not as well understood. A father's poor nutritional status has been shown to have negative consequences on foetal growth and development and ultimately impact the long-term adult health of the offspring. In this study, we examined sperm- and seminal vesicle fluid-mediated mechanisms of preimplantation embryo development alterations in response to sub-optimal paternal diets. RESULTS: Male mice were fed a diet to model either under (low-protein diet (LPD)) or over (high-fat/sugar 'Western' diet (WD)) nutrition, LPD or WD supplemented with methyl donors or a control diet (CD) before mating with age-matched females. Male metabolic health was influenced by WD and MD-WD, with significant changes in multiple serum lipid classes and hepatic 1-carbon metabolites. Sperm RNA sequencing revealed significant changes to mRNA profiles in all groups when compared to CD (LPD: 32, MD-LPD: 17, WD: 53, MD-WD: 35 transcripts). Separate analysis of the seminal vesicle fluid proteome revealed a significant number of differentially expressed proteins in all groups (LPD: 13, MD-LPD: 27, WD: 24, MD-WD: 19) when compared to control. Following mating, in vitro time-lapse imaging of preimplantation embryos revealed a significant increase in the timing of development in all experimental groups when compared to CD embryos. Finally, qPCR analysis of uterine tissue at the time of implantation identified perturbed expression of Cd14 and Ptgs1 following mating with WD-fed males. CONCLUSIONS: Our current study shows that paternal nutritional status has the potential to influence male metabolic and reproductive health, impacting on embryonic development and the maternal reproductive tract. This study highlights potential direct (sperm-mediated) and indirect (seminal vesicle fluid-mediated) pathways in which a father's poor diet could shape the long-term health of his offspring.


Assuntos
Desenvolvimento Embrionário , Hipernutrição , Sêmen , Animais , Masculino , Camundongos , Hipernutrição/fisiopatologia , Feminino , Desnutrição/fisiopatologia , Blastocisto , Espermatozoides
8.
Elife ; 132024 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-39283662

RESUMO

Over the past several decades, a trend toward delayed childbirth has led to increases in parental age at the time of conception. Sperm epigenome undergoes age-dependent changes increasing risks of adverse conditions in offspring conceived by fathers of advanced age. The mechanism(s) linking paternal age with epigenetic changes in sperm remain unknown. The sperm epigenome is shaped in a compartment protected by the blood-testes barrier (BTB) known to deteriorate with age. Permeability of the BTB is regulated by the balance of two mTOR complexes in Sertoli cells where mTOR complex 1 (mTORC1) promotes the opening of the BTB and mTOR complex 2 (mTORC2) promotes its integrity. We hypothesized that this balance is also responsible for age-dependent changes in the sperm epigenome. To test this hypothesis, we analyzed reproductive outcomes, including sperm DNA methylation in transgenic mice with Sertoli cell-specific suppression of mTORC1 (Rptor KO) or mTORC2 (Rictor KO). mTORC2 suppression accelerated aging of the sperm DNA methylome and resulted in a reproductive phenotype concordant with older age, including decreased testes weight and sperm counts, and increased percent of morphologically abnormal spermatozoa and mitochondrial DNA copy number. Suppression of mTORC1 resulted in the shift of DNA methylome in sperm opposite to the shift associated with physiological aging - sperm DNA methylome rejuvenation and mild changes in sperm parameters. These results demonstrate for the first time that the balance of mTOR complexes in Sertoli cells regulates the rate of sperm epigenetic aging. Thus, mTOR pathway in Sertoli cells may be used as a novel target of therapeutic interventions to rejuvenate the sperm epigenome in advanced-age fathers.


Assuntos
Metilação de DNA , Células de Sertoli , Espermatozoides , Masculino , Animais , Células de Sertoli/metabolismo , Camundongos , Espermatozoides/metabolismo , Espermatozoides/fisiologia , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Alvo Mecanístico do Complexo 1 de Rapamicina/genética , Serina-Treonina Quinases TOR/metabolismo , Camundongos Knockout , Alvo Mecanístico do Complexo 2 de Rapamicina/metabolismo , Alvo Mecanístico do Complexo 2 de Rapamicina/genética , Proteína Regulatória Associada a mTOR/metabolismo , Proteína Regulatória Associada a mTOR/genética , Camundongos Transgênicos , Envelhecimento/fisiologia , Envelhecimento/genética , Transdução de Sinais , Proteína Companheira de mTOR Insensível à Rapamicina/metabolismo , Proteína Companheira de mTOR Insensível à Rapamicina/genética , Epigênese Genética
9.
Sci Rep ; 14(1): 21841, 2024 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-39294257

RESUMO

To fertilize eggs, sperm must pass through narrow, complex channels filled with viscoelastic fluids in the female reproductive tract. While it is known that the topography of the surfaces plays a role in guiding sperm movement, sperm have been thought of as swimmers, i.e., their motility comes solely from sperm interaction with the surrounding fluid, and therefore, the surfaces have no direct role in the motility mechanism itself. Here, we examined the role of solid surfaces in the movement of sperm in a highly viscoelastic medium. By visualizing the flagellum interaction with surfaces in a microfluidic device, we found that the flagellum stays close to the surface while the kinetic friction between the flagellum and the surface is in the direction of sperm movement, providing thrust. Additionally, the flow field generated by sperm suggests slippage between the viscoelastic fluid and the solid surface, deviating from the no-slip boundary typically used in standard fluid dynamics models. These observations point to hybrid motility mechanisms in sperm involving direct flagellum-surface interaction in addition to flagellum pushing the fluid. This finding signifies an evolutionary strategy of mammalian sperm crucial for their efficient migration through narrow, mucus-filled passages of the female reproductive tract.


Assuntos
Motilidade dos Espermatozoides , Espermatozoides , Motilidade dos Espermatozoides/fisiologia , Masculino , Animais , Espermatozoides/fisiologia , Viscosidade , Elasticidade , Cauda do Espermatozoide/fisiologia , Feminino
10.
Open Vet J ; 14(8): 2040-2048, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39308734

RESUMO

Background: The preservation of semen quality and kinematic characteristics during cryopreservation is crucial for the reproductive success and genetic management of livestock, particularly in Bali bulls. Aim: This study aimed to investigate the effect of adding purified green tea extract antioxidant Epigallocatechin-3-gallate (EGCG) in tris egg yolk diluent on the quality and kinematic characteristics of frozen semen from Bali bulls. Methods: Fresh and frozen semen samples were obtained from Bali bull and divided into four different treatment groups. P0 contained semen samples + diluent, while P1 to P3 consisted of semen samples + diluent supplemented with EGCG levels of 0.1, 0.15, and 0.2 mg/100 ml, respectively. Data were analyzed using One-way ANOVA and followed by Duncan's test if significant differences were found (p<0.05). Parameters observed included the assessment of fresh semen quality, kinematic analysis, post-thawing sperm viability, and abnormality. Results: The results indicated that the assessment of fresh semen quality showed macroscopic and microscopic semen quality according to SNI 4869-1:2021. Kinematic analysis revealed significant differences in DSL and STR parameters between P0 and P3 (p<0.05). EGCG supplementation also caused significant differences in motility between P0 and P3 (p<0.05). Viability and spermatozoa abnormality with EGCG supplementation did not show significant differences (p>0.05). Conclusion: The best results for motility, kinematics, and sperm morphology variables were found in P1 as it did not exhibit a decrease in motility, kinematics, and sperm morphology. Viability did not show significant differences between P1, P2, and P3, but the best results were found in P2 as it did not exhibit a decrease in viability with mean and standard deviation (66.84 ± 7.88). Abnormality variables also did not show significant differences between P1, P2, and P3, but the best results were found in P2 as it did not exhibit a decrease in abnormality with mean and standard deviation (23.80 ± 7.36).


Assuntos
Antioxidantes , Catequina , Criopreservação , Análise do Sêmen , Preservação do Sêmen , Animais , Masculino , Catequina/análogos & derivados , Catequina/farmacologia , Preservação do Sêmen/veterinária , Criopreservação/veterinária , Antioxidantes/farmacologia , Análise do Sêmen/veterinária , Bovinos , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Chá/química , Fenômenos Biomecânicos/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Sêmen/efeitos dos fármacos , Sêmen/fisiologia , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia
11.
J Biochem Mol Toxicol ; 38(10): e23862, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39318032

RESUMO

Bisphenol A (BPA) is one of the most prevalent endocrine disrupting chemicals (EDCs) and there is widespread concern about the adverse effects of EDCs on human health. However, the exact mechanism of these toxicities has still not been fully deciphered. Additionally, studies have reported the toxicological effects at far low doses to the generally considered no-observed-adverse-effect level (NOAEL) dose. The present study investigates the effects of a sub-acute (28 days) exposure to BPA (10, 50 and 100 mg/kg/day) in adult male mice on various hormones levels, sperm motility, sperm count, functional integrity of sperm plasma membrane, testicular histological changes, oxidative stress markers and DNA damage. The key proteome signatures were quantified by LC-MS/MS analysis using Orbitrap Fusion Lumos Tribrid Mass Spectrometer equipped with nano-LC Easy-nLC 1200. Data suggest that the BPA exposure in all doses (below/above NOAEL dose) have greatly impacted the hormone levels, sperm parameters (sperm count, motility and membrane integrity) and testicular histology. Mass spectrometry-based proteomics data suggested for 1352 differentially expressed proteins (DEPs; 368 upregulated, 984 downregulated) affecting biological process, cellular component, and molecular functions. Specifically searched male reproductive function related proteins suggested a complex network where 46 potential proteins regulating spermatogenesis, sperm structure, activity and membrane integrity while tackling oxidative stress responses were downregulated. These potential biomarkers could shed some more light on our current understanding of the reproductive toxicological effects of BPA and may lead to exploration of novel interventions strategies against these targets for male infertility.


Assuntos
Compostos Benzidrílicos , Fenóis , Proteômica , Testículo , Masculino , Animais , Compostos Benzidrílicos/toxicidade , Fenóis/toxicidade , Camundongos , Testículo/efeitos dos fármacos , Testículo/metabolismo , Testículo/patologia , Proteoma/metabolismo , Proteoma/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Saúde Reprodutiva , Estresse Oxidativo/efeitos dos fármacos
12.
Phys Biol ; 21(6)2024 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-39278237

RESUMO

Rheotaxis is a fundamental mechanism of sperm cells that guides them in navigating towards the oocyte. The present study investigates the phenomenon of sperm rheotaxis in Newtonian and non-Newtonian fluid media, which for the first time explores a viscosity range equivalent to that of the oviductal fluid of the female reproductive tract in rectilinear microfluidic channels. Three parameters, the progressive velocity while performing rheotaxis, the radius of rotation during rheotaxis, and the percentage of rheotactic sperm cells in the bulk and near-wall regions of the microfluidic channel were measured. Numerical simulations of the flow were conducted to estimate the shear rate, flow velocity, and the drag force acting on the sperm head at specific locations where the sperms undergo rheotaxis. Increasing the flow velocity resulted in a change in the position of rheotactic sperm from the bulk center to the near wall region, an increase and subsequent decrease in the sperm's upstream progressive velocity, and a decrease in the radius of rotation. We observed that with an increase in viscosity, rheotactic sperms migrate to the near wall regions at lower flow rates, the upstream progressive velocity of the sperm decreases for Newtonian and increases for non-Newtonian media, and the radius of rotation increases for Newtonian and decreases for non-Newtonian media. These results quantify the effects of fluid properties such as viscosity and flow rate on sperm rheotaxis and navigation, thereby paving the way for manipulating sperm behavior in microfluidic devices, potentially leading to advancements in assisted reproduction techniques.


Assuntos
Motilidade dos Espermatozoides , Espermatozoides , Viscosidade , Espermatozoides/fisiologia , Masculino , Microfluídica/métodos
13.
Narra J ; 4(2): e617, 2024 08.
Artigo em Inglês | MEDLINE | ID: mdl-39280275

RESUMO

Infertility rates have risen significantly, one of which is due to monosodium glutamate (MSG) consumption. Recent studies have shown that flavonoids in black garlic (Allium sativum) act as antioxidants. The aim of this study was to assess the effect of black garlic extract (BGE) on gonadosomatic index, follicle-stimulating hormone (FSH) levels, and spermatozoa quality in rats exposed to MSG. Twenty-five healthy rats, aged ten to twelve weeks, were divided equally into five experimental groups: (1) negative control (NC), no intervention; (2) positive control (PC), fed with MSG 8 mg/kg; and (3) fed with MSG + BGE 200 mg/kg; (4) fed with MSG + BGE 400 mg/kg; and (5) fed with MSG + BGE 600 mg/kg. Oral MSG was administered once a day for two weeks before BGE administration was started for two weeks. The measured endpoints were gonadosomatic index, FSH levels, and spermatozoa concentration and quality (spermatozoa motility and abnormality). Analysis of variance (ANOVA) followed by Duncan's post hoc analysis was used to assess the measurement differences. The result suggested that the administration of BGE did not significantly affect the gonadosomatic index (p=0.513). Significant decreases in FSH levels (p=0.005) and spermatozoa concentration were observed in the PC group compared to other groups (p<0.001). Additionally, spermatozoa motility was significantly lower in the PC group compared to NC, BGE200, BGE400, and BGE600 (p<0.001), with higher motility noted in BGE200, BGE400, and BGE600 compared to PC (p<0.001). Furthermore, PC had significantly higher spermatozoa abnormalities compared to NC, BGE200, BGE400, and BGE600 (p<0.001). In conclusion, administration of BGE had a significant effect on the improvement of FSH levels and the quality of spermatozoa in rats exposed to MSG.


Assuntos
Hormônio Foliculoestimulante , Alho , Extratos Vegetais , Glutamato de Sódio , Espermatozoides , Animais , Masculino , Alho/química , Glutamato de Sódio/farmacologia , Hormônio Foliculoestimulante/sangue , Ratos , Extratos Vegetais/farmacologia , Extratos Vegetais/administração & dosagem , Espermatozoides/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Ratos Wistar , Antioxidantes/farmacologia
14.
Food Funct ; 15(19): 10007-10019, 2024 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-39282919

RESUMO

Background: Perfluorooctane sulfonate (PFOS) is a persistent, widely present environmental pollutant, and its toxicity to male reproduction has gradually attracted attention. Flaxseed oil (FO) is a dietary oil abundant in α-linolenic acid and has been demonstrated to possess multiple health benefits. However, whether FO protects against PFOS-induced testicular injury and its mechanism remain unclear. Methods: C57/BL6 mice were gavaged with different concentrations of FO or PFOS (10 mg kg-1) for 28 days. Blood and testicular tissues were collected for histopathology, proteomics, and biochemical and molecular analyses. Results: Our results showed that FO supplementation significantly attenuated PFOS-induced testicular injury, as indicated by histopathological changes, decreased oxidative stress level, increased sperm count, decreased rate of sperm malformation, and improved functional markers of spermatogenesis. Proteomic analysis showed that differentially expressed proteins were notably enriched in spliceosome pathways. Machine learning algorithms were used to screen the hub gene, and PRPF3 and PUF60 proteins were found to be important for FO to exert protective benefits to testicular injury. Western blot results confirmed that FO supplementation could increase the protein expression of PRPF3 and decrease the protein expression of PUF60 in PFOS-exposed mice. Conclusions: This study revealed that FO can alleviate PFOS-induced testicular dysfunction by regulating RNA alternative splicing. The spliceosome-related proteins PRPF3 and PUF60 may be the potential targets for FO to alleviate PFOS-induced testicular injury. FO supplementation may be an effective dietary intervention to prevent adverse effects of PFOS on testes.


Assuntos
Ácidos Alcanossulfônicos , Processamento Alternativo , Fluorocarbonos , Óleo de Semente do Linho , Camundongos Endogâmicos C57BL , Testículo , Masculino , Animais , Fluorocarbonos/toxicidade , Camundongos , Testículo/efeitos dos fármacos , Testículo/metabolismo , Ácidos Alcanossulfônicos/toxicidade , Processamento Alternativo/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Espermatogênese/efeitos dos fármacos
15.
Sci Rep ; 14(1): 22221, 2024 09 27.
Artigo em Inglês | MEDLINE | ID: mdl-39333615

RESUMO

Gametogenesis produces gametes as a piece of genetic information transmitted to the offspring. While during sexual reproduction, progeny inherits a mix of genetic material from both parents, asexually reproducing organisms transfer a copy of maternal or paternal DNA to the progeny clonally. Parthenogenetic, gynogenetic and hybridogenetic animals have developed various mechanisms of gametogenesis, however, their inheritance is not fully understood. Here, we focused on the inheritance of asexual gametogenesis in hybrid Pelophylax esculentus (RL), emerging after crosses of P. lessonae (LL) and P. ridibundus (RR). To understand the mechanisms of gametogenesis in hybrids, we performed three-generation experiments of sexual P. ridibundus females and hybrids from all-male hybrid populations. Using fluorescent in situ hybridization, micronuclei analysis, flow cytometry and genotyping, we found that most adult hybrid males simultaneously produced two types of clonal sperm. Also, most male tadpole progeny in two successive backcrossed generations simultaneously eliminated L and R parental genomes, while some progeny produced only one type of sperm. We hypothesize that the reproductive variability of males producing two kinds of sperm is an adaptive mechanism to reproduce in mixed populations with P. ridibundus and may explain the extensive distribution of the all-male lineage across the European River Basin.


Assuntos
Hibridização Genética , Reprodução Assexuada , Animais , Masculino , Feminino , Reprodução Assexuada/genética , Padrões de Herança/genética , Espermatozoides/fisiologia , Ranidae/genética , Ranidae/fisiologia , Rana esculenta/genética , Genótipo , Hibridização in Situ Fluorescente
16.
Nat Commun ; 15(1): 8465, 2024 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-39349455

RESUMO

Spermiogenesis, the complex transformation of haploid spermatids into mature spermatozoa, relies on precise spatiotemporal regulation of gene expression at the post-transcriptional level. The mechanisms underlying this critical process remain incompletely understood. Here, we identify centrosomal protein 112 (CEP112) as an essential regulator of mRNA translation during this critical developmental process. Mutations in CEP112 are discovered in oligoasthenoteratospermic patients, and Cep112-deficient male mice recapitulate key phenotypes of human asthenoteratozoospermia. CEP112 localizes to the neck and atypical centrioles of mature sperm and forms RNA granules during spermiogenesis, enriching target mRNAs such as Fsip2, Cfap61, and Cfap74. Through multi-omics analyses and the TRICK reporter assay, we demonstrate that CEP112 orchestrates the translation of target mRNAs. Co-immunoprecipitation and mass spectrometry identify CEP112's interactions with translation-related proteins, including hnRNPA2B1, EEF1A1, and EIF4A1. In vitro, CEP112 undergoes liquid-liquid phase separation, forming condensates that recruit essential proteins and mRNAs. Moreover, variants in patient-derived CEP112 disrupt phase separation and impair translation efficiency. Our results suggest that CEP112 mediates the assembly of RNA granules through liquid-liquid phase separation to control the post-transcriptional expression of fertility-related genes. This study not only clarifies CEP112's role in spermatogenesis but also highlights the role of phase separation in translational regulation, providing insights into male infertility and suggesting potential therapeutic targets.


Assuntos
Biossíntese de Proteínas , Espermatogênese , Masculino , Animais , Espermatogênese/genética , Humanos , Camundongos , RNA Mensageiro/metabolismo , RNA Mensageiro/genética , Fertilidade/genética , Camundongos Knockout , Astenozoospermia/genética , Astenozoospermia/metabolismo , Mutação , Proteínas de Ciclo Celular/metabolismo , Proteínas de Ciclo Celular/genética , Espermatozoides/metabolismo , Espermátides/metabolismo , Oligospermia/genética , Oligospermia/metabolismo , Regulação da Expressão Gênica , Separação de Fases
17.
Nat Commun ; 15(1): 8147, 2024 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-39317727

RESUMO

Modification of guanosine to N7-methylguanosine (m7G) in the variable loop region of tRNA is catalyzed by the METTL1/WDR4 heterodimer and stabilizes target tRNA. Here, we reveal essential functions of Mettl1 in Drosophila fertility. Knockout of Mettl1 (Mettl1-KO) causes no major effect on the development of non-gonadal tissues, but abolishes the production of elongated spermatids and mature sperm, which is fully rescued by expression of a Mettl1-transgene, but not a catalytic-dead Mettl1 transgene. This demonstrates that Mettl1-dependent m7G is required for spermatogenesis. Mettl1-KO results in a loss of m7G modification on a subset of tRNAs and decreased tRNA abundance. Ribosome profiling shows that Mettl1-KO led to ribosomes stalling at codons decoded by tRNAs that were reduced in abundance. Mettl1-KO also significantly reduces the translation efficiency of genes involved in elongated spermatid formation and sperm stability. Germ cell-specific expression of Mettl1 rescues disrupted m7G tRNA modification and tRNA abundance in Mettl1-KO testes but not in non-gonadal tissues. Ribosome stalling is much less detectable in non-gonadal tissues than in Mettl1-KO testes. These findings reveal a developmental role for m7G tRNA modification and indicate that m7G modification-dependent tRNA abundance differs among tissues.


Assuntos
Proteínas de Drosophila , Drosophila melanogaster , Fertilidade , RNA de Transferência , Espermatogênese , Animais , Espermatogênese/genética , Masculino , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , RNA de Transferência/metabolismo , RNA de Transferência/genética , Fertilidade/genética , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/genética , Guanosina/metabolismo , Guanosina/análogos & derivados , Metiltransferases/metabolismo , Metiltransferases/genética , Espermatozoides/metabolismo , Ribossomos/metabolismo , Espermátides/metabolismo , Testículo/metabolismo , Técnicas de Inativação de Genes
18.
Front Endocrinol (Lausanne) ; 15: 1427069, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39324123

RESUMO

Hydrogen sulfide (H2S) is an endogenously produced signaling molecule that belongs to the group of gasotransmitters along with nitric oxide (NO) and carbon monoxide (CO). H2S plays a pivotal role in male reproductive processes. It is produced in various tissues and cells of the male reproductive system, including testicular tissue, Leydig and Sertoli cells, epididymis, seminal plasma, prostate, penile tissues, and sperm cells. This review aims to summarize the knowledge about the presence and effects of H2S in male reproductive tissues and outline possible therapeutic strategies in pathological conditions related to male fertility, e. g. spermatogenetic disorders and erectile dysfunction (ED). For instance, H2S supports spermatogenesis by maintaining the integrity of the blood-testicular barrier (BTB), stimulating testosterone production, and providing cytoprotective effects. In spermatozoa, H2S modulates sperm motility, promotes sperm maturation, capacitation, and acrosome reaction, and has significant cytoprotective effects. Given its vasorelaxant effects, it supports the erection of penile tissue. These findings suggest the importance and therapeutic potential of H2S in male reproduction, paving the way for further research and potential clinical applications.


Assuntos
Sulfeto de Hidrogênio , Reprodução , Espermatogênese , Sulfeto de Hidrogênio/metabolismo , Sulfeto de Hidrogênio/farmacologia , Masculino , Humanos , Animais , Reprodução/efeitos dos fármacos , Reprodução/fisiologia , Espermatogênese/efeitos dos fármacos , Disfunção Erétil/tratamento farmacológico , Disfunção Erétil/metabolismo , Genitália Masculina/metabolismo , Genitália Masculina/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Infertilidade Masculina/metabolismo , Infertilidade Masculina/tratamento farmacológico , Testículo/metabolismo , Testículo/efeitos dos fármacos
19.
Artigo em Inglês | MEDLINE | ID: mdl-39326938

RESUMO

Persons living in industrial environments are exposed to levels of air pollution that can affect their health and fertility. The Czech capital city, Prague, and the Ostrava industrial agglomeration differ in their major sources of air pollution. In Prague, heavy traffic produces high levels of nitrogen oxides throughout the year. In the Ostrava region, an iron industry and local heating are sources of particulate matter (PM) and benzo[a]pyrene (B[a]P), especially in the winter. We evaluated the effects of air pollution on human sperm mitochondrial DNA (mtDNA). Using real-time PCR, we analysed sperm mtDNA copy number and deletion rate in Prague city policemen in two seasons (spring and autumn) and compared the results with those from Ostrava. In Prague, the sperm mtDNA deletion rate was significantly higher in autumn than in spring, which is the opposite of the results from Ostrava. The sperm mtDNA copy number did not show any seasonal differences in either of the cities; it was correlated negatively with sperm concentration, motility, and viability, and with sperm chromatin integrity (assessed with the Sperm Chromatin Structure Assay). The comparison between the two cities showed that the sperm mtDNA deletion rate in spring and the sperm mtDNA copy number in autumn were significantly lower in Prague vs. Ostrava. Our study supports the hypothesis that sperm mtDNA deletion rate is affected by the composition of air pollution. Sperm mtDNA abundance is closely associated with chromatin damage and standard semen characteristics.


Assuntos
Poluição do Ar , Variações do Número de Cópias de DNA , DNA Mitocondrial , Espermatozoides , Humanos , Masculino , DNA Mitocondrial/genética , Espermatozoides/efeitos dos fármacos , República Tcheca , Adulto , Poluição do Ar/efeitos adversos , Estações do Ano , Material Particulado/toxicidade , Material Particulado/efeitos adversos , Poluentes Atmosféricos/toxicidade , Poluentes Atmosféricos/efeitos adversos , Motilidade dos Espermatozoides/efeitos dos fármacos , Polícia
20.
Artigo em Inglês | MEDLINE | ID: mdl-39326943

RESUMO

Diabetes mellitus is known as the "epidemic of the century" due to its global prevalence. Several pre-clinical and clinical studies have shown that male germ cell toxicity is one of the major consequences of diabetes mellitus. Although ß-aminoisobutyric acid (BAIBA) has been shown to be advantageous in the diabetic nephropathy and cardiomyopathy, its specific role in the diabetes-induced testicular toxicity remains unknown. In this study, an attempt was made to elucidate the molecular mechanisms of BAIBA-mediated germ cell protection in diabetic rats. Adult male Sprague-dawley rats were subjected to either no treatment (control) or BAIBA (100 mg/kg; BAIBA control) or Streptozotocin (50 mg/kg; diabetic control) or low (25 mg/kg), medium (50 mg/kg) and high (100 mg/kg) doses of BAIBA in diabetic conditions. Significant alterations in sperm related parameters, oxidative stress and apoptotic biomarkers, pancreatic and testicular histology, DNA damage and changes in expression of proteins in testes were found in the diabetic rats. 100 mg/kg of BAIBA significantly reduced the elevated blood glucose levels (P ≤ 0.05), increased body weight (P ≤ 0.01 in the 4th week), lowered malondialdehyde (P ≤ 0.05) and nitrite levels (P ≤ 0.01), elevated testosterone (P ≤ 0.05) and FSH levels (P ≤ 0.05), increased sperm count and motility (P ≤ 0.01), decreased testicular DNA damage (P ≤ 0.001), improved histological features of pancreas and testes, decreased TUNEL positive cells (P ≤ 0.01), decreased RAGE (P ≤ 0.01) and Bax (P ≤ 0.05) expressions and increased SIRT1 (P ≤ 0.05) and Atg 12 (P ≤ 0.05) expressions in the testes. 50 mg/kg of BAIBA partially restored the above-mentioned parameters whereas 25 mg/kg of BAIBA was found to be insignificant in counteracting the toxicity. It is interesting to note that BAIBA protects male germ cell damage in diabetic rats by regulating the IGF-1/AMPK/SIRT-1 signaling pathway.


Assuntos
Ácidos Aminoisobutíricos , Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 1 , Fator de Crescimento Insulin-Like I , Estresse Oxidativo , Ratos Sprague-Dawley , Transdução de Sinais , Sirtuína 1 , Testículo , Animais , Masculino , Estresse Oxidativo/efeitos dos fármacos , Ratos , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sirtuína 1/metabolismo , Sirtuína 1/genética , Diabetes Mellitus Tipo 1/tratamento farmacológico , Diabetes Mellitus Tipo 1/metabolismo , Testículo/efeitos dos fármacos , Testículo/metabolismo , Testículo/patologia , Ácidos Aminoisobutíricos/farmacologia , Fator de Crescimento Insulin-Like I/metabolismo , Proteínas Quinases Ativadas por AMP/metabolismo , Apoptose/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Dano ao DNA/efeitos dos fármacos
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