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1.
J Assist Reprod Genet ; 39(1): 97-106, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34993708

RESUMO

PURPOSE: To study the morphometric and morphokinetic profiles of pronuclei (PN) between male and female human zygotes. METHOD(S): This retrospective cohort study included 94 consecutive autologous single day 5 transfer cycles leading to a singleton live birth. All oocytes were placed in the EmbryoScope + incubator post-sperm injection with all annotations performed retrospectively by one embryologist (L-SO). Timing parameters included 2nd polar body extrusion (tPB2), sperm-originated PN (tSPNa) or oocyte-originated PN (tOPNa) appearance, and PN fading (tPNF). Morphometrics were evaluated at 8 (stage 1), 4 (stage 2), and 0 h before PNF (stage 3), measuring PN area (um2), PN juxtaposition, and nucleolar precursor bodies (NPB) arrangement. RESULTS: Male zygotes had longer time intervals of tPB2_tSPNa than female zygotes (4.8 ± 0.2 vs 4.2 ± 0.1 h, OR = 1.442, 95% CI 1.009-2.061, p = 0.044). SPN increased in size from stage 1 through 2 to 3 (435.3 ± 7.2, 506.7 ± 8.0, and 556.3 ± 8.9 um2, p = 0.000) and OPN did similarly (399.0 ± 6.1, 464.3 ± 6.7, and 513.8 ± 6.5 um2, p = 0.000), with SPN being significantly larger than OPN at each stage (p < 0.05 respectively). More male than female zygotes reached central PN juxtaposition at stage 1 (76.7% vs 51.0%, p = 0.010), stage 2 (97.7% vs 86.3%, p = 0.048), and stage 3 (97.7% vs 86.3%, p = 0.048). More OPN showed aligned NPBs than in SPN at stage 1 only (44.7% vs 28.7%, p = 0.023). CONCLUSION(S): Embryos with different sexes display different morphokinetic and morphometric features at the zygotic stage. Embryo selection using such parameters may lead to unbalanced sex ratio in resulting offspring.


Assuntos
Oócitos/citologia , Espermatozoides/citologia , Zigoto/citologia , Adulto , Blastômeros/citologia , Blastômeros/microbiologia , Blastômeros/fisiologia , Núcleo Celular/microbiologia , Feminino , Humanos , Modelos Logísticos , Masculino , Oócitos/microbiologia , Estudos Retrospectivos , Espermatozoides/microbiologia , Imagem com Lapso de Tempo/métodos , Zigoto/microbiologia
2.
J Assist Reprod Genet ; 39(1): 19-36, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35034216

RESUMO

Although medical advancements have successfully helped a lot of couples with their infertility by assisted reproductive technologies (ART), sperm selection, a crucial stage in ART, has remained challenging. Therefore, we aimed to investigate novel sperm separation methods, specifically microfluidic systems, as they do sperm selection based on sperm and/or the female reproductive tract (FRT) features without inflicting any damage to the selected sperm during the process. In this review, after an exhaustive studying of FRT features, which can implement by microfluidics devices, the focus was centered on sperm selection and investigation devices. During this study, we tried not to only point to the deficiencies of these systems, but to put forth suggestions for their improvement as well.


Assuntos
Genitália Feminina/fisiologia , Contagem de Espermatozoides/instrumentação , Espermatozoides/citologia , Adulto , Feminino , Genitália Feminina/microbiologia , Humanos , Masculino , Microfluídica/instrumentação , Microfluídica/métodos , Microfluídica/estatística & dados numéricos , Técnicas de Reprodução Assistida/normas , Técnicas de Reprodução Assistida/tendências , Contagem de Espermatozoides/métodos , Contagem de Espermatozoides/tendências , Espermatozoides/microbiologia
3.
Cells ; 11(2)2022 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-35053337

RESUMO

Mammalian sperm differ widely in sperm morphology, and several explanations have been presented to account for this diversity. Less is known about variation in sperm physiology and cellular processes that can give sperm cells an advantage when competing to fertilize oocytes. Capacitation of spermatozoa, a process essential for mammalian fertilization, correlates with changes in motility that result in a characteristic swimming pattern known as hyperactivation. Previous studies revealed that sperm motility and velocity depend on the amount of ATP available and, therefore, changes in sperm movement occurring during capacitation and hyperactivation may involve changes in sperm bioenergetics. Here, we examine differences in ATP levels of sperm from three mouse species (genus Mus), differing in sperm competition levels, incubated under non-capacitating and capacitating conditions, to analyse relationships between energetics, capacitation, and swimming patterns. We found that, in general terms, the amount of sperm ATP decreased more rapidly under capacitating conditions. This descent was related to the development of a hyperactivated pattern of movement in two species (M. musculus and M. spicilegus) but not in the other (M. spretus), suggesting that, in the latter, temporal dynamics and energetic demands of capacitation and hyperactivation may be decoupled or that the hyperactivation pattern differs. The decrease in ATP levels during capacitation was steeper in species with higher levels of sperm competition than in those with lower levels. Our results suggest that, during capacitation, sperm consume more ATP than under non-capacitating conditions. This higher ATP consumption may be linked to higher velocity and lateral head displacement, which are associated with hyperactivated motility.


Assuntos
Metabolismo Energético , Capacitação Espermática/fisiologia , Espermatozoides/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Sobrevivência Celular , Masculino , Camundongos , Motilidade Espermática/fisiologia , Espermatozoides/citologia , Fatores de Tempo
4.
Cell Mol Life Sci ; 79(2): 91, 2022 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-35072818

RESUMO

Mitochondria tailor their morphology to execute their specialized functions in different cell types and/or different environments. During spermatogenesis, mitochondria undergo continuous morphological and distributional changes with germ cell development. Deficiencies in these processes lead to mitochondrial dysfunction and abnormal spermatogenesis, thereby causing male infertility. In recent years, mitochondria have attracted considerable attention because of their unique role in the regulation of piRNA biogenesis in male germ cells. In this review, we describe the varied characters of mitochondria and focus on key mitochondrial factors that play pivotal roles in the regulation of spermatogenesis, from primordial germ cells to spermatozoa, especially concerning metabolic shift, stemness and reprogramming, mitochondrial transformation and rearrangement, and mitochondrial defects in human sperm. Further, we discuss the molecular mechanisms underlying these processes.


Assuntos
Diferenciação Celular , Células Germinativas/citologia , Infertilidade Masculina/patologia , Mitocôndrias/fisiologia , Doenças Mitocondriais/fisiopatologia , Espermatozoides/citologia , Animais , Humanos , Infertilidade Masculina/etiologia , Infertilidade Masculina/metabolismo , Masculino
5.
Cells ; 11(2)2022 01 11.
Artigo em Inglês | MEDLINE | ID: mdl-35053349

RESUMO

Sperm competition is thought to impose strong selection on males to produce competitive ejaculates to outcompete rival males under competitive mating conditions. Our understanding of how different sperm traits influence fertilization success, however, remains limited, especially in wild populations. Recent literature highlights the importance of incorporating multiple ejaculate traits and pre-copulatory sexually selected traits in analyses aimed at understanding how selection acts on sperm traits. However, variation in a male's ability to gain fertilization success may also depend upon a range of social and ecological factors that determine the opportunity for mating events both within and outside of the social pair-bond. Here, we test for an effect of sperm quantity and sperm size on male reproductive success in the red-back fairy-wren (Malurus melanocephalus) while simultaneously accounting for pre-copulatory sexual selection and potential socio-ecological correlates of male mating success. We found that sperm number (i.e., cloacal protuberance volume), but not sperm morphology, was associated with reproductive success in male red-backed fairy-wrens. Most notably, males with large numbers of sperm available for copulation achieved greater within-pair paternity success. Our results suggest that males use large sperm numbers as a defensive strategy to guard within-pair paternity success in a system where there is a high risk of sperm competition and female control of copulation. Finally, our work highlights the importance of accounting for socio-ecological factors that may influence male mating opportunities when examining the role of sperm traits in determining male reproductive success.


Assuntos
Animais Selvagens/fisiologia , Passeriformes/fisiologia , Contagem de Espermatozoides , Animais , Animais Selvagens/genética , Cloaca , Feminino , Humanos , Masculino , Modelos Biológicos , Passeriformes/genética , Fenótipo , Espermatozoides/citologia
6.
Fertil Steril ; 117(1): 64-73, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34548170

RESUMO

OBJECTIVE: To investigate the intraindividual agreement of the sperm chromatin dispersion (SCD) assay results to assess sperm DNA fragmentation (SDF) in men with infertility. DESIGN: Diagnostic test reliability study. SETTING: Andrology laboratories. PATIENT(S): A total of 219 men with infertility. INTERVENTION(S): Sperm DNA fragmentation assessment in two ejaculates of the same subjects within a 3-month interval, using the SCD assay performed and analyzed by the same observers under similar testing conditions. MAIN OUTCOME MEASURE(S): Cohen's κ statistics to assess the degree of agreement between the pairs of results after converting the nominal SCD values into categorical data, that is, normal (<20%), intermediate (21%-29%), and high (≥30%) SDF rates. We also assessed the pairs of results using reliability measures for numerical variables (intraclass correlation coefficient for consistency using the two-way mixed-effects model and Bland-Altman plots). RESULT(S): The degree of agreement in classifying patients according to normal and pathological SDF classes was overall substantial (κ = 0.632; 95% confidence interval [CI], 0.546-0.718). A total of 76.7% of individuals were classified under the same class using paired ejaculates. The agreement rate was highest (approximately 80%) in ejaculates initially classified as either normal or high and lowest (approximately 60%) among those with intermediate SDF levels. The frequency of intermediate SDF ejaculates downgraded to normal or upgrade to high SDF classes in the second test was similar (approximately 20%). The intraclass correlation coefficient was 0.856 (95% CI, 0.812-0.887), and the mean difference between the pairs of observations was 0.80% (95% CI, -0.72 to 2.23), indicating no systematic difference between paired observations. CONCLUSION(S): Our study indicates a substantial intraindividual agreement of paired SCD assay results to classify men with infertility into three SDF categories: normal, intermediate, and high. The reliability of the SCD assay was adequate and exceeded 0.80 using two ejaculates analyzed within a 3-month interval under similar conditions. Although this evidence overall supports a single SCD test for patient classification using predefined SDF thresholds, particularly when the first test shows normal or high SDF levels, one in four men will have discordant values in paired ejaculates. Clinicians should be judicious when using SDF test results in decision-making.


Assuntos
Fragmentação do DNA , Infertilidade Masculina/diagnóstico , Análise do Sêmen/métodos , Adulto , Andrologia/métodos , Brasil , Cromatina/química , Cromatina/metabolismo , DNA/análise , DNA/metabolismo , Humanos , Infertilidade Masculina/genética , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Espanha , Espermatozoides/química , Espermatozoides/citologia , Espermatozoides/metabolismo
7.
Physiol Rev ; 102(1): 7-60, 2022 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33880962

RESUMO

The spermatozoon is a highly differentiated and polarized cell, with two main structures: the head, containing a haploid nucleus and the acrosomal exocytotic granule, and the flagellum, which generates energy and propels the cell; both structures are connected by the neck. The sperm's main aim is to participate in fertilization, thus activating development. Despite this common bauplan and function, there is an enormous diversity in structure and performance of sperm cells. For example, mammalian spermatozoa may exhibit several head patterns and overall sperm lengths ranging from ∼30 to 350 µm. Mechanisms of transport in the female tract, preparation for fertilization, and recognition of and interaction with the oocyte also show considerable variation. There has been much interest in understanding the origin of this diversity, both in evolutionary terms and in relation to mechanisms underlying sperm differentiation in the testis. Here, relationships between sperm bauplan and function are examined at two levels: first, by analyzing the selective forces that drive changes in sperm structure and physiology to understand the adaptive values of this variation and impact on male reproductive success and second, by examining cellular and molecular mechanisms of sperm formation in the testis that may explain how differentiation can give rise to such a wide array of sperm forms and functions.


Assuntos
Exocitose/fisiologia , Interações Espermatozoide-Óvulo/fisiologia , Espermatozoides/fisiologia , Testículo/citologia , Animais , Evolução Biológica , Humanos , Masculino , Mamíferos/fisiologia , Espermatozoides/citologia
8.
Int J Mol Sci ; 22(23)2021 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-34884637

RESUMO

The adverse effects of radiation are proportional to the total dose and dose rate. We aimed to investigate the effects of radiation dose rate on different organs in mice. The mice were subjected to low dose rate (LDR, ~3.4 mGy/h) and high dose rate (HDR, ~51 Gy/h) radiation. LDR radiation caused severe tissue toxicity, as observed in the histological analysis of testis. It adversely influenced sperm production, including sperm count and motility, and induced greater sperm abnormalities. The expression of markers of early stage spermatogonial stem cells, such as Plzf, c-Kit, and Oct4, decreased significantly after LDR irradiation, compared to that following exposure of HDR radiation, in qPCR analysis. The compositional ratios of all stages of spermatogonia and meiotic cells, except round spermatid, were considerably reduced by LDR in FACS analysis. Therefore, LDR radiation caused more adverse testicular damage than that by HDR radiation, contrary to the response observed in other organs. Therefore, the dose rate of radiation may have differential effects, depending on the organ; it is necessary to evaluate the effect of radiation in terms of radiation dose, dose rate, organ type, and other conditions.


Assuntos
Espermatogênese/efeitos da radiação , Testículo/efeitos da radiação , Animais , Relação Dose-Resposta à Radiação , Raios gama , Masculino , Camundongos , Modelos Animais , Doses de Radiação , Espermátides/citologia , Espermátides/efeitos da radiação , Espermatogônias/citologia , Espermatogônias/efeitos da radiação , Espermatozoides/citologia , Espermatozoides/efeitos da radiação , Testículo/citologia
9.
Bull Exp Biol Med ; 172(2): 254-262, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34855079

RESUMO

We studied the effect of extracellular vesicles of the follicular fluid on morphofunctional characteristics of human spermatozoa using CASA (computer-assisted sperm analysis) analytical system. The vesicles were obtained by sequential centrifugation at different rotational speeds and frozen at -80°C in the Sydney IVF Gamete Buffer medium. The sperm fraction was isolated from the seminal fluid of 21 patients aged 27-36 years by differential centrifugation in a density gradient. The precipitate was suspended in Sydney IVF Gamete Buffer to a concentration of 106/ml and incubated with vesicles (1:2) at 37°C in a CO2 incubator for 30 min and 1 h. Sperm fraction incubated without vesicles served as the control. After incubation, some sperm samples were centrifuged at 700g for 5 min and fixed in 2.5% glutaraldehyde in 0.1 M buffer for transmission electron microscopy. After 30-min and 1-h incubation, the progressive and total sperm motility improved, the curvilinear and linear velocity of spermatozoa did not change significantly. Incubation with vesicles significantly changed the trajectory of sperm movement, which can attest to an increase in their hyperactivation and, probably, fertilizing capacity. Analysis of the effect of extracellular vesicles of follicular fluid on sperm motility will help to improve the effectiveness of assisted reproductive technology programs with male infertility factor by improving sperm characteristics in patients with asthenozoospermia and increasing the fertilizing ability of the sperm.


Assuntos
Vesículas Extracelulares/fisiologia , Líquido Folicular/citologia , Espermatozoides/fisiologia , Acrossomo/metabolismo , Acrossomo/fisiologia , Adulto , Vesículas Extracelulares/metabolismo , Feminino , Regulação da Expressão Gênica , Humanos , Técnicas In Vitro , Masculino , Análise do Sêmen , Transdução de Sinais/genética , Motilidade Espermática/fisiologia , Espermatozoides/citologia
10.
Cell Mol Life Sci ; 79(1): 50, 2021 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-34936029

RESUMO

Circular RNA (circRNA) biogenesis requires a backsplicing reaction, promoted by inverted repeats in cis-flanking sequences and trans factors, such as RNA-binding proteins (RBPs). Among these, FUS plays a key role. During spermatogenesis and sperm maturation along the epididymis such a molecular mechanism has been poorly explored. With this in mind, we chose circCNOT6L as a study case and wild-type (WT) as well as cannabinoid receptor type-1 knock-out (Cb1-/-) male mice as animal models to analyze backsplicing mechanisms. Our results suggest that spermatozoa (SPZ) have an endogenous skill to circularize mRNAs, choosing FUS as modulator of backsplicing and under CB1 stimulation. A physical interaction between FUS and CNOT6L as well as a cooperation among FUS, RNA Polymerase II (RNApol2) and Quaking (QKI) take place in SPZ. Finally, to gain insight into FUS involvement in circCNOT6L biogenesis, FUS expression was reduced through RNA interference approach. Paternal transmission of FUS and CNOT6L to oocytes during fertilization was then assessed by using murine unfertilized oocytes (NF), one-cell zygotes (F) and murine oocytes undergoing parthenogenetic activation (PA) to exclude a maternal contribution. The role of circCNOT6L as an active regulator of zygote transition toward the 2-cell-like state was suggested using the Embryonic Stem Cell (ESC) system. Intriguingly, human SPZ exactly mirror murine SPZ.


Assuntos
RNA Circular/metabolismo , Proteína FUS de Ligação a RNA/metabolismo , Ribonucleases/genética , Espermatozoides , Animais , Feminino , Humanos , Masculino , Camundongos , Camundongos Knockout , Oócitos , Espermatozoides/citologia , Espermatozoides/metabolismo , Zigoto/metabolismo
11.
Cells ; 10(12)2021 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-34943823

RESUMO

We present a new method for the selection of individual sperm cells using a microfluidic device that automatically traps each cell in a separate microdroplet that then individually self-assembles with other microdroplets, permitting the controlled measurement of the cells using quantitative phase microscopy. Following cell trapping and droplet formation, we utilize quantitative phase microscopy integrated with bright-field imaging for individual sperm morphology and motility inspection. We then perform individual sperm selection using a single-cell micromanipulator, which is enhanced by the microdroplet-trapping procedure described above. This method can improve sperm selection for intracytoplasmic sperm injection, a common type of in vitro fertilization procedure.


Assuntos
Fertilização In Vitro , Microscopia , Espermatozoides/citologia , Movimento Celular , Humanos , Masculino , Microfluídica
12.
Nat Commun ; 12(1): 6839, 2021 11 25.
Artigo em Inglês | MEDLINE | ID: mdl-34824237

RESUMO

Mammalian male germ cell development is a stepwise cell-fate transition process; however, the full-term developmental profile of male germ cells remains undefined. Here, by interrogating the high-precision transcriptome atlas of 11,598 cells covering 28 critical time-points, we demonstrate that cell-fate transition from mitotic to post-mitotic primordial germ cells is accompanied by transcriptome-scale reconfiguration and a transitional cell state. Notch signaling pathway is essential for initiating mitotic arrest and the maintenance of male germ cells' identities. Ablation of HELQ induces developmental arrest and abnormal transcriptome reprogramming of male germ cells, indicating the importance of cell cycle regulation for proper cell-fate transition. Finally, systematic human-mouse comparison reveals potential regulators whose deficiency contributed to human male infertility via mitotic arrest regulation. Collectively, our study provides an accurate and comprehensive transcriptome atlas of the male germline cycle and allows for an in-depth understanding of the cell-fate transition and determination underlying male germ cell development.


Assuntos
Espermatozoides/citologia , Espermatozoides/crescimento & desenvolvimento , Animais , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Masculino , Meiose/genética , Camundongos , Mitose/genética , Receptores Notch/genética , Receptores Notch/metabolismo , Transdução de Sinais , Espermatogênese/genética , Espermatozoides/metabolismo , Transcriptoma
13.
Int J Mol Sci ; 22(22)2021 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-34830255

RESUMO

Ketamine, which used to be widely applied in human and animal medicine as a dissociative anesthetic, has become a popular recreational drug because of its hallucinogenic effect. Our previous study preliminarily proved that ketamine could inhibit human sperm function by affecting intracellular calcium concentration ([Ca2+]i). However, the specific signaling pathway of [Ca2+]i induced by ketamine in human sperm is still not clear yet. Here, the N-methyl-d-aspartic acid (NMDA) receptor was detected in the tail region of human sperm. Its physiological ligand, NMDA (50 µM), could reverse ketamine's inhibitory effect on human sperm function, and its antagonist, MK801 (100 µM), could restrain the effect of NMDA. The inhibitory effect caused by 4 mM ketamine or 100 µM MK801 on [Ca2+]i, which is a central factor in the regulation of human sperm function, could also be recovered by 50 µM NMDA. The results suggest that the NMDA receptor is probably involved in the inhibitory effect of ketamine on human sperm functions.


Assuntos
Anestésicos Dissociativos/farmacologia , Ketamina/farmacologia , N-Metilaspartato/farmacologia , Receptores de N-Metil-D-Aspartato/genética , Espermatozoides/efeitos dos fármacos , Adulto , Cálcio/metabolismo , Células Cultivadas , Maleato de Dizocilpina/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Expressão Gênica , Humanos , Transporte de Íons/efeitos dos fármacos , Masculino , N-Metilaspartato/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Espermatozoides/citologia , Espermatozoides/metabolismo
14.
STAR Protoc ; 2(4): 100933, 2021 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-34806046

RESUMO

The preservation of mammalian freeze-dried (FD) spermatozoa is commonly performed using small glass ampules; however, they are bulky and breakable. In this study, we present a protocol to prepare and preserve mouse FD sperm using thin plastic sheets. This approach allows storing thousands of mouse strains in a card folder. We can also send the FD sperm domestically using a postcard without any extra equipment. For complete details on the use and execution of this protocol, please refer to Ito et al. (2021).


Assuntos
Liofilização/métodos , Preservação do Sêmen/métodos , Espermatozoides , Animais , Masculino , Camundongos , Espermatozoides/citologia , Espermatozoides/fisiologia
15.
Cell Mol Life Sci ; 78(24): 8049-8071, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34748024

RESUMO

Multiple RNA pathways are required to produce functional sperm. Here, we review RNA post-transcriptional regulation during spermatogenesis with particular emphasis on the role of 3' end modifications. From early studies in the 1970s, it became clear that spermiogenesis transcripts could be stored for days only to be translated at advanced stages of spermatid differentiation. The transition between the translationally repressed and active states was observed to correlate with the shortening of the transcripts' poly(A) tail, establishing a link between RNA 3' end metabolism and male germ cell differentiation. Since then, numerous RNA metabolic pathways have been implicated not only in the progression through spermatogenesis, but also in the maintenance of genomic integrity. Recent studies have characterized the elusive 3' biogenesis of Piwi-interacting RNAs (piRNAs), identified a critical role for messenger RNA (mRNA) 3' uridylation in meiotic progression, established the mechanisms that destabilize transcripts with long 3' untranslated regions (3'UTRs) in post-mitotic cells, and defined the physiological relevance of RNA exonucleases and deadenylases in male germ cells. In this review, we discuss RNA processing in the male germline in the light of the most recent findings. A brief recollection of different RNA-processing events will aid future studies exploring post-transcriptional regulation in spermatogenesis.


Assuntos
Regulação da Expressão Gênica , Processamento Pós-Transcricional do RNA , Espermatogênese , Espermatozoides/fisiologia , Animais , Humanos , Masculino , Espermatozoides/citologia
16.
Int J Mol Sci ; 22(21)2021 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-34769240

RESUMO

In mammals, integrins are heterodimeric transmembrane glycoproteins that represent a large group of cell adhesion receptors involved in cell-cell, cell-extracellular matrix, and cell-pathogen interactions. Integrin receptors are an important part of signalization pathways and have an ability to transmit signals into and out of cells and participate in cell activation. In addition to somatic cells, integrins have also been detected on germ cells and are known to play a crucial role in complex gamete-specific physiological events, resulting in sperm-oocyte fusion. The main aim of this review is to summarize the current knowledge on integrins in reproduction and deliver novel perspectives and graphical interpretations presenting integrin subunits localization and their dynamic relocation during sperm maturation in comparison to the oocyte. A significant part of this review is devoted to discussing the existing view of the role of integrins during sperm migration through the female reproductive tract; oviductal reservoir formation; sperm maturation processes ensuing capacitation and the acrosome reaction, and their direct and indirect involvement in gamete membrane adhesion and fusion leading to fertilization.


Assuntos
Integrinas/metabolismo , Oócitos/metabolismo , Capacitação Espermática , Interações Espermatozoide-Óvulo/fisiologia , Espermatozoides/metabolismo , Animais , Feminino , Humanos , Masculino , Oócitos/citologia , Espermatozoides/citologia
17.
Zoolog Sci ; 38(5): 444-450, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34664919

RESUMO

The genus Mesobiotus was separated from the genus Macrobiotus in 2016 and the name referred to its phylogenetic position among the family Macrobiotidae; however, knowledge of the reproductive behavior of this genus is limited compared to those of Paramacrobiotus and Macrobiotus. This study comprehensively provides the reproductive traits, including the gamete morphologies and behavioral observations, of Mesobiotus. The morphology of its spermatozoon showed a length that was intermediary among those of Paramacrobiotus and Macrobiotus species. The sequence of mating behavior was generally conserved in the three species of Macrobiotidae. They showed the described five steps observed in Paramacrobiotus and Macrobiotus; however, the males of Mesobiotus repeated ejaculations in a mating session, which is the first observation of premature ejaculation in tardigrades. Our results indicated that Mesobiotus has the potential to be a model to show the linkage between genera with respect to the morphology and behavior in the family Macrobiotidae.


Assuntos
Comportamento Sexual Animal , Tardígrados/anatomia & histologia , Tardígrados/fisiologia , Animais , Ejaculação , Feminino , Masculino , Óvulo/citologia , Espermatozoides/citologia , Tardígrados/classificação
18.
Genes (Basel) ; 12(10)2021 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-34680969

RESUMO

This study aimed to characterize the protein composition of fractionated seminal plasma (SP) by liquid chromatography mass spectrometry (LC-MS/MS) analysis and investigate its effects on survival of frozen-thaw (FT) boar spermatozoa following storage. Seminal plasma (SP) was fractionated by gel filtration chromatography to give two fractions, SP1 with more than 40 kDa (>40 kDa) and SP2 with less than 40 kDa (<40 kDa). SP1 and SP2 were subjected to LC-MS/MS and bioinformatics analysis. Following cryopreservation, FT boar semen (n = 7) was thawed in Beltsville Thawing Solution (BTS), BTS + SP1 or BTS + SP2, stored at different periods and subjected to post-thaw (PT) quality assessment. A total of 52 and 22 abundant proteins were detected in SP1 and SP2, respectively. FN1, ANGPTL1, and KIF15 proteins were more abundance in SP1, whereas a high abundance of spermadhesins (PSP-I and PSP-II) was detected in SP2. Proteins of the fractionated SP were involved in various biological processes, such as cell motility and signal transduction. The dominant pathway of SP1 proteins was the apelin signaling pathway (GNA13, MEF2D, SPHK2, and MEF2C), whereas a pathway related to lysosome (CTSH, CTSB, and NPC2) was mainly represented by SP2 proteins. In most of the boars, significantly higher motility characteristics, membrane integrity, and viability were observed in FT spermatozoa exposed to SP1 or SP2 compared with BTS. The results of our study confirm that a combination of several proteins from the fractionated SP exerted beneficial effects on the sperm membrane, resulting in improved quality characteristics following PT storage.


Assuntos
Proteínas/genética , Motilidade Espermática/genética , Espermatozoides/citologia , Sus scrofa/genética , Animais , Cromatografia Líquida , Criopreservação , Congelamento , Masculino , Sêmen/citologia , Sêmen/metabolismo , Análise do Sêmen/métodos , Preservação do Sêmen , Espermatozoides/crescimento & desenvolvimento , Sus scrofa/crescimento & desenvolvimento , Suínos/genética , Espectrometria de Massas em Tandem
19.
Int J Mol Sci ; 22(19)2021 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-34639209

RESUMO

The process of freezing cells or tissues and depositing them in liquid nitrogen at -196 °C is called cryopreservation. Sub-zero temperature is not a physiological condition for cells and water ice crystals represent the main problem since they induce cell death, principally in large cells like oocytes, which have a meiotic spindle that degenerates during this process. Significantly, cryopreservation represents an option for fertility preservation in patients who develop gonadal failure for any condition and those who want to freeze their germ cells for later use. The possibility of freezing sperm, oocytes, and embryos has been available for a long time, and in 1983 the first birth with thawed oocytes was achieved. From the mid-2000s forward, the use of egg vitrification through intracytoplasmic sperm injection has improved pregnancy rates. Births using assisted reproductive technologies (ART) have some adverse conditions and events. These risks could be associated with ART procedures or related to infertility. Cryopreservation generates changes in the epigenome of gametes and embryos, given that ART occurs when the epigenome is most vulnerable. Furthermore, cryoprotective agents induce alterations in the integrity of germ cells and embryos. Notably, cryopreservation extensively affects cell viability, generates proteomic profile changes, compromises crucial cellular functions, and alters sperm motility. This technique has been widely employed since the 1980s and there is a lack of knowledge about molecular changes. The emerging view is that molecular changes are associated with cryopreservation, affecting metabolism, cytoarchitecture, calcium homeostasis, epigenetic state, and cell survival, which compromise the fertilization in ART.


Assuntos
Cálcio/metabolismo , Criopreservação/normas , Embrião de Mamíferos/citologia , Epigênese Genética , Células Germinativas/citologia , Infertilidade/terapia , Proteoma/metabolismo , Sobrevivência Celular , Crioprotetores/química , Feminino , Preservação da Fertilidade/normas , Fertilização In Vitro , Células Germinativas/metabolismo , Humanos , Infertilidade/metabolismo , Infertilidade/patologia , Masculino , Oócitos/citologia , Oócitos/metabolismo , Gravidez , Espermatozoides/citologia , Espermatozoides/metabolismo
20.
Int J Mol Sci ; 22(17)2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34502430

RESUMO

The fertilization of freshwater fish occurs in an environment that may negatively affect the gametes; therefore, the specific mechanisms triggering the encounters of gametes would be highly expedient. The egg and ovarian fluid are likely the major sources of these triggers, which we confirmed here for rainbow trout (Oncorhynchus mykiss). The ovarian fluid affected significantly spermatozoa performance: it supported high velocity for a longer period and changed the motility pattern from tumbling in water to straightforward moving in the ovarian fluid. Rainbow trout ovarian fluid induced a trapping chemotaxis-like effect on activated male gametes, and this effect depended on the properties of the activating medium. The interaction of the spermatozoa with the attracting agents was accompanied by the "turn-and-run" behavior involving asymmetric flagellar beating and Ca2+ concentration bursts in the bent flagellum segment, which are characteristic of the chemotactic response. Ovarian fluid created the optimal environment for rainbow trout spermatozoa performance, and the individual peculiarities of the egg (ovarian fluid)-sperm interaction reflect the specific features of the spawning process in this species.


Assuntos
Quimiotaxia/fisiologia , Fertilização/fisiologia , Oncorhynchus mykiss/metabolismo , Ovário/metabolismo , Espermatozoides/metabolismo , Zigoto/metabolismo , Animais , Sinalização do Cálcio/fisiologia , Feminino , Masculino , Ovário/citologia , Espermatozoides/citologia , Zigoto/citologia
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