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1.
Int J Mol Sci ; 22(6)2021 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-33806823

RESUMO

Sperm motility is linked to the activation of signaling pathways that trigger movement. These pathways are mainly dependent on Ca2+, which acts as a secondary messenger. The maintenance of adequate Ca2+ concentrations is possible thanks to proper concentrations of other ions, such as K+ and Na+, among others, that modulate plasma membrane potential and the intracellular pH. Like in every cell, ion homeostasis in spermatozoa is ensured by a vast spectrum of ion channels supported by the work of ion pumps and transporters. To achieve success in fertilization, sperm ion channels have to be sensitive to various external and internal factors. This sensitivity is provided by specific channel structures. In addition, novel sperm-specific channels or isoforms have been found with compositions that increase the chance of fertilization. Notably, the most significant sperm ion channel is the cation channel of sperm (CatSper), which is a sperm-specific Ca2+ channel required for the hyperactivation of sperm motility. The role of other ion channels in the spermatozoa, such as voltage-gated Ca2+ channels (VGCCs), Ca2+-activated Cl-channels (CaCCs), SLO K+ channels or voltage-gated H+ channels (VGHCs), is to ensure the activation and modulation of CatSper. As the activation of sperm motility differs among metazoa, different ion channels may participate; however, knowledge regarding these channels is still scarce. In the present review, the roles and structures of the most important known ion channels are described in regard to regulation of sperm motility in animals.


Assuntos
Ativação do Canal Iônico , Canais Iônicos/química , Canais Iônicos/metabolismo , Motilidade Espermática , Espermatozoides/fisiologia , Animais , Cálcio/metabolismo , Canais de Cálcio/química , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Sinalização do Cálcio , Canais de Cloreto/química , Canais de Cloreto/genética , Canais de Cloreto/metabolismo , Humanos , Canais Iônicos/genética , Masculino , Canais de Potássio/química , Canais de Potássio/genética , Canais de Potássio/metabolismo , Canais de Sódio/química , Canais de Sódio/genética , Canais de Sódio/metabolismo , Relação Estrutura-Atividade
2.
J Vis Exp ; (168)2021 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-33645583

RESUMO

Recording of the electrical activity from one of the smallest cells of a mammalian organism- a sperm cell- has been a challenging task for electrophysiologists for many decades. The method known as "spermatozoan patch clamp" was introduced in 2006. It has enabled the direct recording of ion channel activity in whole-cell and cell-attached configurations and has been instrumental in describing sperm cell physiology and the molecular identity of various calcium, potassium, sodium, chloride, and proton ion channels. However, recording from single spermatozoa requires advanced skills and training in electrophysiology. This detailed protocol summarizes the step-by-step procedure and highlights several 'tricks-of-the-trade' in order to make it available to anyone who wishes to explore the fascinating physiology of the sperm cell. Specifically, the protocol describes recording from human and murine sperm cells but can be adapted to essentially any mammalian sperm cell of any species. The protocol covers important details of the application of this technique, such as isolation of sperm cells, selection of reagents and equipment, immobilization of the highly motile cells, formation of the tight (Gigaohm) seal between a recording electrode and the plasma membrane of the sperm cells, transition into the whole-spermatozoan mode (also known as break-in), and exemplary recordings of the sperm cell calcium ion channel, CatSper, from six mammalian species. The advantages and limitations of the sperm patch clamp method, as well as the most critical steps, are discussed.


Assuntos
Membrana Celular/fisiologia , Fenômenos Eletrofisiológicos , Espermatozoides/fisiologia , Animais , Cálcio/metabolismo , Membrana Celular/efeitos dos fármacos , Tamanho Celular , Dissecação , Fenômenos Eletrofisiológicos/efeitos dos fármacos , Flagelos/efeitos dos fármacos , Flagelos/fisiologia , Humanos , Concentração de Íons de Hidrogênio , Transporte de Íons/efeitos dos fármacos , Macaca mulatta , Masculino , Camundongos Endogâmicos C57BL , Técnicas de Patch-Clamp , Perfusão , Progesterona/farmacologia , Soluções , Espermatozoides/citologia , Espermatozoides/efeitos dos fármacos
3.
Medicine (Baltimore) ; 100(10): e24828, 2021 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-33725837

RESUMO

OBJECTIVE: To assess the association of conventional semen parameters and sperm DNA fragmentation with risk of recurrent spontaneous abortion (RSA). DESIGN: Systematic review and meta-analysis. SETTING: Not applicable. PATIENTS: Total 1,690 male partners of women with RSA, and 1,337 male partners of fertile control women. INTERVENTIONS: Case-control or cohort studies were determined by searching PubMed, Google Scholar, Cochrane Libraries, China Biology Medicine disc, Chinese Scientific Journals Fulltext Database, China National Knowledge Infrastructure, and Wanfang Database. RSA was defined as two or more previous pregnancy losses. The fertile women refer to the reproductive women who have had at least a normal pregnancy history and no history of abortion. MAIN OUTCOME MEASURES: This study included eight outcome measures: semen volume(ml), semen pH value, sperm density(106/ml), sperm viability (%), sperm progressive motility rate (%), normal sperm morphology rate (%), sperm deformity rate(%), sperm DNA fragmentation index (DFI) (%). The summary measures were reported as standardized mean difference (SMD) with 95% confidence interval (CI). RESULTS: Finally, twenty-four studies were included for analysis. Overall, male partners of women with RSA had a significantly lower level of sperm density (SMD = -0.53, 95%CI: - 0.75 to -0.30), sperm viability (SMD = -1.03, 95%CI: - 1.52 to -0.54), sperm progressive motility rate (SMD = -0.76, 95%CI:-1.06 - -0.46), and normal sperm morphology rate (SMD =  -0.56, 95%CI: - 0.99 to -0.12), and had a significantly higher rate of sperm deformity rate (SMD = 1.29, 95%CI: 0.60 - 1.97), and sperm DFI (SMD = 1.60, 95%CI: 1.04 to 2.17), when compared with the reference group. However, there were no statistically significant differences for semen volume (SMD = -0.03, 95%CI: -0.14 - 0.08) and semen pH value (SMD =  -0.23, 95% CI: -0.50 to 0.05) among 2 groups. CONCLUSIONS: The results of this analysis support an association of sperm density, sperm viability, sperm progressive motility rate, normal sperm morphology rate, sperm deformity rate, as well as sperm DFI with RSA. However, given the significant heterogeneity between studies and the lack of more detailed data on the subjects, further large-scale prospective studies are needed.


Assuntos
Aborto Habitual/etiologia , Sêmen/fisiologia , Espermatozoides/fisiologia , Aborto Habitual/etnologia , Grupo com Ancestrais do Continente Asiático , Estudos de Casos e Controles , Fragmentação do DNA , Feminino , Humanos , Concentração de Íons de Hidrogênio , Masculino , Fatores de Risco , Contagem de Espermatozoides , Motilidade Espermática , Espermatozoides/citologia
4.
Poult Sci ; 100(2): 1221-1238, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33518080

RESUMO

High levels of polyunsaturated fatty acids in avian sperm cause more susceptibility to lipid peroxidation. Aging in roosters reduces the antioxidant capacity of sperm and thus fertility. The purpose of this study was to investigate the effects of different levels of alpha-lipoic acid (ALA) as a feed supplement to improve the semen quality and fertility parameters of aged broiler breeder roosters and identification of its most effective level. A total of forty-two roosters at 45 wk of age were randomly assigned to 7 treatments (0, 15, 40, 70, 95, 120, and 145 mg ALA/bird per day) for 8 wk. Semen parameters and body weight were assessed biweekly, and testosterone plasma levels were determined in the 8th wk of the experimental period. Artificial insemination was performed at the end of the experiment to evaluate the fertility potential. The dietary administration of ALA had no significant effects on body weight, semen volume, average path velocity, linearity, straightness, wobble, the amplitude of lateral head displacement, beat-cross frequency, sperm concentration, morphology, plasma testosterone level, fertility, or hatchability (P > 0.05). Alpha-lipoic acid supplementations resulted in a significant decrease in seminal malondialdehyde concentration and immotile (type D) sperms (P < 0.05). The total motility, progressive motility (types A + type B sperms), curvilinear velocity, straight-line velocity, viability, and membrane integrity of sperm improved with ALA dietary supplementations (P < 0.05). With increasing ALA levels, improvement in semen parameters had an incremental trend until the level of 95 mg ALA. Thus, 95 mg dietary ALA as an antioxidant supplement can improve semen quality of aging breeder roosters while higher doses resulted in no further improvement.


Assuntos
Envelhecimento/fisiologia , Galinhas/fisiologia , Fertilidade/efeitos dos fármacos , Sêmen/fisiologia , Ácido Tióctico/administração & dosagem , Ração Animal/análise , Animais , Peso Corporal/fisiologia , Suplementos Nutricionais , Feminino , Fertilidade/fisiologia , Inseminação Artificial/veterinária , Masculino , Sêmen/química , Análise do Sêmen/veterinária , Motilidade Espermática , Espermatozoides/fisiologia
5.
Methods Mol Biol ; 2218: 29-35, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33606220

RESUMO

Fish sperm show many measurable parameters which react sensitively in a dose- and time-dependent way to toxic exposure. Fish sperm is therefore used as an in vitro toxicology test system. One of the most sensitive and easily detectable parameters is progressive motility which can be measured by a computer-assisted sperm analysis (CASA) system. Here we describe a simple protocol to test the effect of environmental toxicants by using zebrafish (Danio rerio) sperm.


Assuntos
Substâncias Perigosas/toxicidade , Processamento de Imagem Assistida por Computador/métodos , Análise do Sêmen/métodos , Espermatozoides/fisiologia , Peixe-Zebra/fisiologia , Animais , Computadores , Masculino , Motilidade Espermática/efeitos dos fármacos , Motilidade Espermática/fisiologia , Espermatozoides/efeitos dos fármacos
6.
Nat Commun ; 12(1): 666, 2021 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-33531493

RESUMO

Temperature has a crucial influence on the places where species can survive and reproduce. Past research has primarily focused on survival, making it unclear if temperature fluctuations constrain reproductive success, and if so whether populations harbour the potential to respond to climatic shifts. Here, using two decades of data from a large experimental breeding programme of the iconic ostrich (Struthio camelus) in South Africa, we show that the number of eggs females laid and the number of sperm males produced were highly sensitive to natural temperature extremes (ranging from -5 °C to 45 °C). This resulted in reductions in reproductive success of up to 44% with 5 °C deviations from their thermal optimum. In contrast, gamete quality was largely unaffected by temperature. Extreme temperatures also did not expose trade-offs between gametic traits. Instead, some females appeared to invest more in reproducing at high temperatures, which may facilitate responses to climate change. These results show that the robustness of fertility to temperature fluctuations, and not just temperature increases, is a critical aspect of species persistence in regions predicted to undergo the greatest change in climate volatility.


Assuntos
Fertilidade/fisiologia , Temperatura , Animais , Feminino , Temperatura Alta , Masculino , África do Sul , Espermatozoides/fisiologia
7.
Aquat Toxicol ; 231: 105740, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33440272

RESUMO

Ensuring that oocytes are fertilized by a single sperm during broadcast spawning is crucial for the fertilization success of many marine invertebrates. Although the adverse impacts of ocean acidification (OA) on various marine species have been revealed in recent years, its impact on polyspermy and the underlying mechanisms involved remain largely unknown. Therefore, in the present study, the effect of OA on polyspermy risk was assessed in a broadcast spawning bivalve, Tegillarca granosa. In addition, the impacts of OA on the two polyspermy blocking processes, the fast block (membrane depolarization) and the permanent block (cortical reaction), were investigated. The results show that the exposure of oocytes to two future OA scenarios (pH 7.8 and pH 7.4) leads to significant increases in polyspermy risk, about 1.70 and 2.38 times higher than the control, respectively. The maximum change in the membrane potential during oocyte membrane depolarization markedly decreased to 15.79 % (pH 7.8) and 34.06 % (pH 7.4) of the control value. Moreover, the duration of oocyte membrane depolarization was significantly reduced to approximately 63.38 % (pH 7.8) and 21.91 % (pH 7.4) of the control. In addition, cortical granule exocytosis, as well as microfilament migration, were significantly arrested by OA treatment. Exposure to future OA scenarios also led to significant reductions in the ATP and Ca2+ content of the oocytes, which may explain the hampered polyspermy blocking. Overall, the present study suggests that OA may significantly increase polyspermy risk in T. granosa by inhibiting membrane depolarization and arresting cortical granule exocytosis.


Assuntos
Ácidos/química , Bivalves/fisiologia , Grânulos Citoplasmáticos/metabolismo , Exocitose , Potenciais da Membrana/fisiologia , Oceanos e Mares , Espermatozoides/fisiologia , Citoesqueleto de Actina/efeitos dos fármacos , Citoesqueleto de Actina/metabolismo , Trifosfato de Adenosina/farmacologia , Animais , Cálcio/metabolismo , Movimento Celular/efeitos dos fármacos , Grânulos Citoplasmáticos/efeitos dos fármacos , Exocitose/efeitos dos fármacos , Feminino , Fertilização/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Masculino , Potenciais da Membrana/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Poluentes Químicos da Água/toxicidade
8.
Lancet Haematol ; 8(2): e149-e160, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33513374

RESUMO

As survival rates in teenagers and young adults diagnosed with haematological malignancies now exceed 70%, it is important that long-term quality of life, including measures to protect future fertility, are considered and discussed with patients and their families. Although discussion on the effect of planned cancer treatment on fertility is standard of care, knowledge of potential fertility treatment options and when they should be offered in haematological malignancies is not always so clear. In each case, the advice on the appropriate preservation of fertility depends upon a complex interplay of factors, weighing out the risk of future infertility against the risk of fertility preservation treatment, and recommendations must be made on a case-by-case basis. The aim of this Review is to evaluate the gonadotoxicity of treatments of prevalent haematological malignancies in teenagers and young adults, and provide an evidence-based framework to help with fertility discussion and management at the time of diagnosis, relapse or resistant disease, and in long-term follow-up settings.


Assuntos
Preservação da Fertilidade/métodos , Neoplasias Hematológicas/terapia , Antineoplásicos/uso terapêutico , Feminino , Congelamento , Neoplasias Hematológicas/tratamento farmacológico , Neoplasias Hematológicas/radioterapia , Humanos , Masculino , Óvulo/fisiologia , Espermatozoides/fisiologia , Irradiação Corporal Total
9.
Int J Mol Sci ; 22(1)2021 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-33406808

RESUMO

Polyamines are ubiquitous polycationic compounds that are highly charged at physiological pH. While passing through the epididymis, sperm lose their capacity to synthesize the polyamines and, upon ejaculation, again come into contact with the polyamines contained in the seminal fluid, unleashing physiological events that improve sperm motility and capacitation. In the present work, we hypothesize about the influence of polyamines, namely, spermine, spermidine, and putrescine, on the activity of sperm channels, evaluating the intracellular concentrations of chloride [Cl-]i, calcium [Ca2+]i, sodium [Na+]i, potassium [K+]i, the membrane Vm, and pHi. The aim of this is to identify the possible regulatory mechanisms mediated by the polyamines on sperm-specific channels under capacitation and non-capacitation conditions. The results showed that the presence of polyamines did not directly influence the activity of calcium and chloride channels. However, the results suggested an interaction of polyamines with sodium and potassium channels, which may contribute to the membrane Vm during capacitation. In addition, alkalization of the pHi revealed the possible activation of sperm-specific Na+/H+ exchangers (NHEs) by the increased levels of cyclic AMP (cAMP), which were produced by soluble adenylate cyclase (sAC) and interact with the polyamines, evidence that is supported by in silico analysis.


Assuntos
Canais Iônicos/fisiologia , Poliaminas/farmacologia , Capacitação Espermática/efeitos dos fármacos , Motilidade Espermática/efeitos dos fármacos , Espermatozoides/fisiologia , Animais , Cálcio/metabolismo , AMP Cíclico/metabolismo , Canais Iônicos/efeitos dos fármacos , Masculino , Potenciais da Membrana , Camundongos , Potássio/metabolismo , Espermatozoides/efeitos dos fármacos
10.
Am J Hum Genet ; 108(2): 309-323, 2021 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-33472045

RESUMO

Asthenoteratozoospermia characterized by multiple morphological abnormalities of the flagella (MMAF) has been identified as a sub-type of male infertility. Recent progress has identified several MMAF-associated genes with an autosomal recessive inheritance in human affected individuals, but the etiology in approximately 40% of affected individuals remains unknown. Here, we conducted whole-exome sequencing (WES) and identified hemizygous missense variants in the X-linked CFAP47 in three unrelated Chinese individuals with MMAF. These three CFAP47 variants were absent in human control population genome databases and were predicted to be deleterious by multiple bioinformatic tools. CFAP47 encodes a cilia- and flagella-associated protein that is highly expressed in testis. Immunoblotting and immunofluorescence assays revealed obviously reduced levels of CFAP47 in spermatozoa from all three men harboring deleterious missense variants of CFAP47. Furthermore, WES data from an additional cohort of severe asthenoteratozoospermic men originating from Australia permitted the identification of a hemizygous Xp21.1 deletion removing the entire CFAP47 gene. All men harboring hemizygous CFAP47 variants displayed typical MMAF phenotypes. We also generated a Cfap47-mutated mouse model, the adult males of which were sterile and presented with reduced sperm motility and abnormal flagellar morphology and movement. However, fertility could be rescued by the use of intra-cytoplasmic sperm injections (ICSIs). Altogether, our experimental observations in humans and mice demonstrate that hemizygous mutations in CFAP47 can induce X-linked MMAF and asthenoteratozoospermia, for which good ICSI prognosis is suggested. These findings will provide important guidance for genetic counseling and assisted reproduction treatments.


Assuntos
Astenozoospermia/genética , Infertilidade Masculina/genética , Animais , Astenozoospermia/patologia , Astenozoospermia/fisiopatologia , Estudos de Coortes , Feminino , Deleção de Genes , Genes Ligados ao Cromossomo X , Hemizigoto , Humanos , Infertilidade Masculina/metabolismo , Infertilidade Masculina/patologia , Infertilidade Masculina/fisiopatologia , Masculino , Camundongos Endogâmicos C57BL , Mutação , Mutação de Sentido Incorreto , Linhagem , Fenótipo , Injeções de Esperma Intracitoplásmicas , Motilidade Espermática , Cauda do Espermatozoide/ultraestrutura , Espermatozoides/patologia , Espermatozoides/fisiologia , Espermatozoides/ultraestrutura , Sequenciamento Completo do Exoma
11.
PLoS One ; 16(1): e0244648, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33417615

RESUMO

Honey bee (Apis mellifera) queens have a remarkable organ, the spermatheca, which successfully stores sperm for years after a virgin queen mates. This study uniquely characterized and quantified the transcriptomes of the spermathecae from mated and virgin honey bee queens via RNA sequencing to identify differences in mRNA levels based on a queen's mating status. The transcriptome of drone semen was analyzed for comparison. Samples from three individual bees were independently analyzed for mated queen spermathecae and virgin queen spermathecae, and three pools of semen from ten drones each were collected from three separate colonies. In total, the expression of 11,233 genes was identified in mated queen spermathecae, 10,521 in virgin queen spermathecae, and 10,407 in drone semen. Using a cutoff log2 fold-change value of 2.0, we identified 212 differentially expressed genes between mated and virgin spermathecal queen tissues: 129 (1.4% of total) were up-regulated and 83 (0.9% of total) were down-regulated in mated queen spermathecae. Three genes in mated queen spermathecae, three genes in virgin queen spermathecae and four genes in drone semen that were more highly expressed in those tissues from the RNA sequencing data were further validated by real time quantitative PCR. Among others, expression of Kielin/chordin-like and Trehalase mRNAs was highest in the spermathecae of mated queens compared to virgin queen spermathecae and drone semen. Expression of the mRNA encoding Alpha glucosidase 2 was higher in the spermathecae of virgin queens. Finally, expression of Facilitated trehalose transporter 1 mRNA was greatest in drone semen. This is the first characterization of gene expression in the spermathecae of honey bee queens revealing the alterations in mRNA levels within them after mating. Future studies will extend to other reproductive tissues with the purpose of relating levels of specific mRNAs to the functional competence of honey bee queens and the colonies they head.


Assuntos
Abelhas/genética , Transcriptoma , Animais , Abelhas/fisiologia , Feminino , Genes de Insetos , Inseminação , Masculino , Reprodução , Sêmen/fisiologia , Comportamento Sexual Animal , Espermatozoides/fisiologia
12.
Trop Anim Health Prod ; 53(1): 97, 2021 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-33415455

RESUMO

Induction of heat stress as an experimental procedure in animals is commonly used to examine heat-related impacts on sperm quality. This study aimed to develop potential heat stress models that could be used at any time of the year, to advance the study of seasonal infertility in the pig under controlled conditions. Heat stress was induced by either housing boars (n = 6) at 30 °C inside a hot room for 42 days (55-65% humidity; LD 12:12 h; in vivo), or by heating boar semen (n = 7) for 30 min at various temperatures (35.5, 38.8, 40, 42, 46, 50, 54 and 60 °C; in vitro). Sperm motility was then characterized by computer-assisted sperm analysis (CASA; IVOS version 10: Hamilton Thorne, USA), and DNA integrity was evaluated by terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) and flow cytometry. Our in vivo hot room model induced biologically meaningful levels of DNA damage in boar spermatozoa (10.1 ± 1.9 hot room vs. 6.7 ± 1.7% control; P > 0.05), although not statistically significant from controls. Moreover, sperm concentration and motility parameters did not differ between treatments (P > 0.05). Compared to the 38.8 °C control, our in vitro heat shock model significantly increased sperm DNA damage after incubation at 54 and 60 °C (3.0 ± 1.0, 2.9 ± 1.0, 1.2 ± 0.3, 2.5 ± 0.7, 9.0 ± 3.7, 16.2 ± 7.1, 14.2 ± 5.8 and 41.8 ± 18.6% respectively; P ≤ 0.05). However, these temperatures rendered sperm completely immotile or dead, with most motility parameters declining rapidly to zero above 40 or 42 °C. In conclusion, our results suggest that temperature combined with individual factors may contribute to a boar's overall susceptibility to heat stress. Refinement of these models particularly of the in vitro heat shock model could be further pursued to overcome environmental variability, reduce whole animal experiments and provide a putative diagnostic fertility screening tool to evaluate heat tolerance in the boar.


Assuntos
Resposta ao Choque Térmico/fisiologia , Espermatozoides/fisiologia , Sus scrofa/fisiologia , Animais , Masculino , Modelos Animais
13.
Open Biol ; 11(1): 200347, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33465325

RESUMO

Coronavirus disease 2019 (COVID-19) has emerged as a new public health crisis, threatening almost all aspects of human life. Originating in bats, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is transmitted to humans through unknown intermediate hosts, where it is primarily known to cause pneumonia-like complications in the respiratory system. Organ-to-organ transmission has not been ruled out, thereby raising the possibility of the impact of SARS-CoV-2 infection on multiple organ systems. The male reproductive system has been hypothesized to be a potential target of SARS-CoV-2 infection, which is supported by some preliminary evidence. This may pose a global threat to male fertility potential, as men are more prone to SARS-CoV-2 infection than women, especially those of reproductive age. Preliminary reports have also indicated the possibility of sexual transmission of SARS-CoV-2. It may cause severe complications in infected couples. This review focuses on the pathophysiology of potential SARS-CoV-2 infection in the reproductive organs of males along with their invasion mechanisms. The risks of COVID-19 on male fertility as well as the differences in vulnerability to SARS-CoV-2 infection compared with females have also been highlighted.


Assuntos
/patologia , Saúde Reprodutiva , /patogenicidade , /imunologia , Citocinas/metabolismo , Fragmentação do DNA , Humanos , Linfócitos/metabolismo , Linfócitos/virologia , Masculino , Estresse Oxidativo , Espermatozoides/fisiologia , Espermatozoides/virologia
14.
Methods Mol Biol ; 2202: 93-102, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32857349

RESUMO

Reactive oxygen species (ROS) could have a negative impact on sperm cellular function and viability. This chapter describes a protocol for oxidative stress evaluation using dichlorofluorescein (DCF) which can specifically reveal intracellular reactive oxygen species. The protocol described here has been used in human and teleost species sperm samples. The method can be used with two approaches: (1) flow cytometry, for quantification of DCF+ cells, or (2) confocal microscopy, for the localization of ROS within the cells.


Assuntos
Microscopia Confocal/métodos , Espécies Reativas de Oxigênio/análise , Espermatozoides/metabolismo , Animais , Peixes , Citometria de Fluxo/métodos , Fluoresceínas/química , Humanos , Peróxido de Hidrogênio/metabolismo , Masculino , Estresse Oxidativo/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Espermatozoides/fisiologia
15.
Chemosphere ; 262: 127880, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32777607

RESUMO

BACKGROUND: Bisphenol A (BPA) is a well-known endocrine disruptor that affects male fertility. However, the main biological events through which BPA affects spermatogenesis remain to be identified. METHODS: Adult male mice were treated by feeding with drinking water containing BPA (0.2 µg/ml, 20 µg/ml, 200 µg/ml, respectively) for two months. Testes were collected for protein extraction or for immunohistochemical analysis. Epididymal spermatozoa were collected for sperm quality evaluation and male fertility assay by in vitro fertility (IVF). Serums were collected for detection of testosterone levels. Proteins associated with germ cell proliferation, meiosis, blood-testis barrier, and steroidogenesis production were examined in BPA-treated and control mice testes. CCK8 assay was used to detect the effect of BPA on the proliferation of GC-1 and GC-2 cells. RESULTS: The BPA-treated mice were characterized by decreased sperm quality, serum testosterone levels and, sub-fertile phenotype characterizing with low pregnancy rates and reduced fertilization efficiency. In lower BPA (0.2 µg/ml) treatment, PCNA and PLZF were down-expressed that indicated impaired germ cell proliferation. SYCP3 was down-expressed in BPA-treated mice, but expressions of other proteins associated with meiosis and blood-testis barrier were not significantly altered. CYP11A1 and HSD3B1 were down-expressed in BPA-treated mice that demonstrated reduced steroidogenesis activity. BPA has a concentration-dependent inhibition effect on the proliferation of GC-1 and GC-2 cells. Conclusively, low doses BPA exposure reduced mice sperm quality mainly by impairing germ cell proliferation, leading to reduced male fertility. The study would provide relevant information for investigation on molecular mechanisms and protective strategy on male production.


Assuntos
Compostos Benzidrílicos/toxicidade , Disruptores Endócrinos/toxicidade , Fenóis/toxicidade , Espermatozoides/fisiologia , Animais , Proliferação de Células/efeitos dos fármacos , Disruptores Endócrinos/metabolismo , Epididimo/efeitos dos fármacos , Feminino , Fertilidade/efeitos dos fármacos , Masculino , Camundongos , Gravidez , Espermatogênese/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Testículo/efeitos dos fármacos , Testes de Toxicidade Crônica
16.
Ecotoxicol Environ Saf ; 207: 111563, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33254417

RESUMO

Exposure to endocrine-disrupting chemicals (EDCs) has been hypothesized as a cause of declining sheep reproductive efficiency. Understanding the long-term effects of EDCs such as heavy metals on reproductive health requires investigation in 'real life' of sheep that are reared in industrial areas. The aim of this study was to evaluate the effect of long-term exposure of Kermani rams to high levels of environmental heavy metals probably emitted from a copper smelter at KhatoonAbad in ShahreBabak, Kerman province. Testicular characteristics were determined in randomly-selected rams (3-4 years old) at 4 directions (south, north, east, and west) and 4 distances (10, 20, 30, and 40 km) from the smelter. Testicular trace element contents, size, serum testosterone, histological attributes and seminal characteristics, except semen volume, were affected by both the direction and the distance from the smelter (P < 0.05). Testicular contents of Pb, Cd, Cr, and Ni, and sperm abnormalities were higher at 10 km south from the smelter and lower at 40 km west. Other parameters were higher at 40 km west and lower at 10 km south. Interestingly, the testicular contents of Cu at 10 km south were lower and associated with higher sperm abnormalities in the rams reared closer to the smelter. The highest weight, length and circumference of the testis were found at 40 km west. The lowest concentration of testosterone was observed at 10 km south, being 92.6% lower than the highest values obtained at 40 km west. The diameter of seminiferous tubules and epithelial height at 10 km south were 8.9% and 27.5% lower than the highest values obtained at 40 km west. A positive correlation between Pb, Cd, Cr and Ni contents in the testis with sperm abnormalities, and a negative correlation between these elements with the other parameters were found. It was concluded that long-term exposure to heavy metals might have been a cause of decreased fertility in rams and probably other living species in this region.


Assuntos
Poluentes Ambientais/toxicidade , Metais Pesados/toxicidade , Testículo/efeitos dos fármacos , Animais , Cobre/toxicidade , Masculino , Reprodução , Análise do Sêmen/veterinária , Ovinos , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Testículo/fisiologia , Testosterona/sangue , Testes de Toxicidade Crônica , Oligoelementos/metabolismo
17.
Ecotoxicol Environ Saf ; 207: 111219, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32931966

RESUMO

Contamination by organic and inorganic compounds remains one of the most complex problems in both brackish and marine environments, causing potential implications for the reproductive success and survival of several broadcast spawners. Ficopomatus enigmaticus is a tubeworm polychaete that has previously been used as a model organism for ecotoxicological analysis, due to its sensitivity and ecological relevance. In the present study, the effects of five trace elements (zinc, copper, cadmium, arsenic and lead), one surfactant (sodium dodecyl sulfate, SDS) and one polycyclic aromatic hydrocarbon (benzo(a)pyrene, B(a)P) on the sperm quality of F. enigmaticus were investigated. Sperm suspensions were exposed in vitro to different concentrations of each selected contaminant under four salinity conditions (10, 20, 30, 35). Possible adverse effects on sperm function were assessed by measuring oxidative stress, membrane integrity, viability and DNA damage. Sperm quality impairments induced by organic contaminants were more evident than those induced by inorganic compounds. SDS exerted the largest effect on sperm. In addition, F. enigmaticus sperm showed high tolerance to salinity variation, supporting the wide use of this species as a promising model organism for ecotoxicological assays. Easy and rapid methods on polychaete spermatozoids were shown to be effective as integrated sperm quality parameters or as an alternative analysis for early assessment of marine and brackish water pollution.


Assuntos
Poliquetos/fisiologia , Espermatozoides/fisiologia , Poluentes Químicos da Água/toxicidade , Animais , Arsênico/farmacologia , Bioensaio , Cádmio/toxicidade , Ecotoxicologia/métodos , Masculino , Poliquetos/efeitos dos fármacos , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Salinidade , Oligoelementos/toxicidade
18.
Pesqui. vet. bras ; 40(12): 1048-1053, Dec. 2020. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1155032

RESUMO

Awareness of the physiological changes that occur when animals are subjected to climatic changes that are considered stressful is essential to maintain animal welfare and to be able to exploit their reproductive potential efficiently and rationally. The present study was carried out to evaluate climatic variables' influence on physiological parameters, and Murrah buffalo ejaculates reared in a humid tropical climate in the Amazon. The immediate analyzes pertinent to the physical and morphological characteristics of the ejaculates were carried out and corresponded in the rainy season (RS) volume of 3.4±2.0mL; the mass activity of 4.4±0.5; motility of 80.4±5.6%; vigor of 4.4±0.4; concentration of 657,300±237,865.1 x 106sptz/mL; major defects of 9.0±2.6%; minor defects of 11.2±3.9%; total defects 20.2±5.3% and sperm plasma membrane integrity (SPMI) 84.8±5.6%, whereas in the non-rainy season (nRS), the results were 4.0±2.1mL; the mass activity of 3.0±1.0; motility of 56.2±13.4%; vigor of 3.0±1.0; concentration of 586,000±291,925.9 x 106sptz/mL; major defects of 20.8±9.9%; minor defects of 27.5±6.3%; total defects 48.3±9.3% and SPMI of 57.9±12.4%. Furthermore, a statistical difference (P<0.05) was observed for the parameters mass activity, motility, vigor, major defects, minor defects, total defects, and sperm plasma membrane integrity between both periods. The data on heart frequency, superficial temperature (head, back, groin, and scrotal pouch) showed a statistical difference between both periods (P<0.05). To conclude is necessary specific management in the non-rainy season that thermal stress is not a determining factor in reducing the reproductive quality of buffaloes; it is necessary to use means to improve animal welfare; one alternative is to use baths regularly for these animals or provide constant access to areas of rivers or lakes, as well as shading, preventing the buffaloes from being directly exposed to the unfavorable thermal environment.(AU)


O conhecimento das alterações fisiológicas que ocorrem quando os animais são submetidos a alterações climatológicas consideradas estressantes é fundamental para manter o bem-estar animal, e poder explorar o seu potencial reprodutivo de forma eficiente e racional. O presente estudo foi realizado com o objetivo de avaliar a influência de variáveis climáticas sobre parâmetros fisiológicos e de ejaculados de búfalos, da raça Murrah, criados em clima tropical úmido da Amazônia. As análises imediatas pertinentes às características físicas e morfológicas dos ejaculados foram realizadas e corresponderam no período chuvoso (PCh) o volume de 3,4±2,0mL, turbilhonamento de 4,4±0,5; motilidade de 80,4±5,6%; vigor de 4,4±0,4; concentração de 657.300±237.865,1 x 106sptz/mL; defeitos maiores de 9,0±2,6%; defeitos menores de 11,2±3,9%; defeitos totais de 20,2±5,3% e integridade da membrana plasmática (IMP) de 84,8±5,6%, enquanto que no período não chuvoso (PnCh), os resultados foram de 4,0±2,1mL; turbilhonamento de 3,0±1,0; motilidade de 56,2±13,4%; vigor de 3,0±1,0; concentração de 586.000±291.925,9 x 106sptz/mL; defeitos maiores de 20,8±9,9%; defeitos menores de 27,5±6,3%; defeitos totais de 48,3±9,3% e IMP de 57,9±12,4%. Observou-se diferença estatística (P<0,05) para os parâmetros movimento de massa, motilidade, vigor, defeitos maiores, defeitos menores, defeitos totais e integridade da membrana plasmática entre os dois períodos. Dados de frequência cardíaca, temperatura superficial (cabeça, dorso, virilha e bolsa escrotal) diferiram estatisticamente entre os períodos (P<0,05). Conclui-se que se faz necessário usar de um manejo específico no período não chuvoso para que o estresse térmico não seja um fator determinante na redução da qualidade reprodutiva dos búfalos, para isto se faz necessário utilizar de meios para melhorar o bem-estar animal, sendo uma das alternativas fazer uso de banhos regularmente para estes animais, ou disponibilizar acesso constante destes a áreas de rios ou lagos, assim como sombreamentos, evitando que os búfalos fiquem expostos diretamente ao ambiente térmico desfavorável.(AU)


Assuntos
Espermatozoides/fisiologia , Búfalos/fisiologia , Análise do Sêmen/veterinária , Bem-Estar do Animal , Parâmetros
20.
Anim Sci J ; 91(1): e13493, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33314533

RESUMO

Although successful fertilization is completed by only 150 sperm in the pig oviduct, more than 50,000 sperms are required to achieve a fertilization rate of more than 70% by pig in vitro fertilization (IVF). In this study, to improve the efficiency of pig IVF, the effects of hypoxic conditions and treatment with creatine and methyl-beta cyclodextrin (MßCD) on the glycolytic pathway were investigated. Under low O2 conditions, zig-zag motility was strongly induced within 30 min; however, the induction disappeared at 60 min. Although caffeine suppressed zig-zag motility under low O2 conditions, creatine induced and sustained zig-zag motility until 120 min. Additionally, pretreatment with MßCD for 15 min greatly enhanced zig-zag motility via ATP production in sperm incubated with creatine under low O2 conditions. Sperm pretreated with MßCD were used for IVF in medium containing creatine under low O2 conditions. A fertilization rate of approximately 70% was achieved with only 1.0 x 104 sperms/mL, and there were few polyspermic embryos. Therefore, our novel method was beneficial for efficient production of pig embryos in vitro. Moreover, the zig-zag motility may be a novel movement which boar capacitated sperm exhibit in the culture medium.


Assuntos
Anaerobiose/fisiologia , Creatina/farmacologia , Fertilização In Vitro/métodos , Fertilização In Vitro/veterinária , Fertilização/efeitos dos fármacos , Motilidade Espermática , Interações Espermatozoide-Óvulo/efeitos dos fármacos , Espermatozoides/fisiologia , Suínos/fisiologia , beta-Ciclodextrinas/farmacocinética , Animais , Sinergismo Farmacológico , Ejaculação/fisiologia , Feminino , Fertilização/fisiologia , Masculino , Interações Espermatozoide-Óvulo/fisiologia , beta-Ciclodextrinas/farmacologia
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