RESUMO
Teratozoospermia is a significant cause of male infertility, but the pathogenic mechanism of acephalic spermatozoa syndrome (ASS), one of the most severe teratozoospermia, remains elusive. We previously reported Spermatogenesis Associated 6 (SPATA6) as the component of the sperm head-tail coupling apparatus (HTCA) required for normal assembly of the sperm head-tail conjunction, but the underlying molecular mechanism has not been explored. Here, we find that the co-chaperone protein BAG5, expressed in step 9-16 spermatids, is essential for sperm HTCA assembly. BAG5-deficient male mice show abnormal assembly of HTCA, leading to ASS and male infertility, phenocopying SPATA6-deficient mice. In vivo and in vitro experiments demonstrate that SPATA6, cargo transport-related myosin proteins (MYO5A and MYL6) and dynein proteins (DYNLT1, DCTN1, and DNAL1) are misfolded upon BAG5 depletion. Mechanistically, we find that BAG5 forms a complex with HSPA8 and promotes the folding of SPATA6 by enhancing HSPA8's affinity for substrate proteins. Collectively, our findings reveal a novel protein-regulated network in sperm formation in which BAG5 governs the assembly of the HTCA by activating the protein-folding function of HSPA8.
Assuntos
Proteínas do Citoesqueleto , Infertilidade Masculina , Teratozoospermia , Tiazóis , Animais , Humanos , Masculino , Camundongos , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Dineínas/metabolismo , Proteínas de Choque Térmico HSC70/genética , Proteínas de Choque Térmico HSC70/metabolismo , Infertilidade Masculina/genética , Infertilidade Masculina/patologia , Chaperonas Moleculares/genética , Chaperonas Moleculares/metabolismo , Dobramento de Proteína , Sêmen/metabolismo , Cabeça do Espermatozoide/fisiologia , Espermatogênese/genética , Espermatozoides/metabolismo , Teratozoospermia/metabolismo , Teratozoospermia/patologiaRESUMO
The perinuclear theca (PT) is a cytoskeletal element encapsulating the sperm nucleus; however, our understanding of the physiological roles of PT in sperm is very limited. We show that Calicin interacts with itself and many other PT components, indicating it may serve as an organizing center of the PT assembly. Calicin is detectable first when surrounding the acrosome, then detected around the entire nucleus, and finally translocated to the postacrosomal region of spermatid heads. Intriguingly, loss of Calicin specifically causes surface subsidence of sperm heads in the nuclear condensation stage. Calicin interacts with inner acrosomal membrane (IAM) protein Spaca1 and nuclear envelope (NE) components to form an "IAM-PT-NE" structure. Intriguingly, Ccin-knockout sperm also exhibit DNA damage and failure of fertilization. Our study provides solid animal evidence to suggest that the PT encapsulating sperm nucleus helps shape the sperm head and maintain the nuclear structure.
Assuntos
Proteínas do Citoesqueleto , Sêmen , Cabeça do Espermatozoide , Animais , Proteínas do Citoesqueleto/metabolismo , Citoesqueleto/fisiologia , Masculino , Camundongos , Sêmen/metabolismo , Cabeça do Espermatozoide/metabolismo , Cabeça do Espermatozoide/fisiologiaRESUMO
Mammals exhibit a tremendous amount of variation in sperm morphology and despite the acknowledgement of sperm structural diversity across taxa, its functional significance remains poorly understood. Of particular interest is the sperm of rodents. While most Eutherian mammal spermatozoa are relatively simple cells with round or paddle-shaped heads, rodent sperm are often more complex and, in many species, display a striking apical hook. The function of the sperm hook remains largely unknown, but it has been hypothesized to have evolved as an adaptation to inter-male sperm competition and thus has been implicated in increased swimming efficiency or in the formation of collective sperm movements. Here we empirically test these hypotheses within a single lineage of Peromyscus rodents, in which closely related species naturally vary in their mating systems, sperm head shapes, and propensity to form sperm aggregates of varying sizes. We performed sperm morphological analyses as well as in vitro analyses of sperm aggregation and motility to examine whether the sperm hook (i) morphologically varies across these species and (ii) associates with sperm competition, aggregation, or motility. We demonstrate inter-specific variation in the sperm hook and then show that hook width negatively associates with sperm aggregation and sperm swimming speed, signifying that larger hooks may be a hindrance to sperm movement within this group of mice. Finally, we confirmed that the sperm hook hinders motility within a subset of Peromyscus leucopus mice that spontaneously produced sperm with no or highly abnormal hooks. Taken together, our findings suggest that any adaptive value of the sperm hook is likely associated with a function other than inter-male sperm competition, such as interaction with ova or cumulous cells during fertilization, or migration through the complex female reproductive tract.
Assuntos
Evolução Biológica , Peromyscus/fisiologia , Cabeça do Espermatozoide/fisiologia , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/química , Espermatozoides/fisiologia , Animais , Masculino , Camundongos , Peromyscus/anatomia & histologiaRESUMO
Reproductive success depends on efficient sperm movement driven by axonemal dynein-mediated microtubule sliding. Models predict sliding at the base of the tail - the centriole - but such sliding has never been observed. Centrioles are ancient organelles with a conserved architecture; their rigidity is thought to restrict microtubule sliding. Here, we show that, in mammalian sperm, the atypical distal centriole (DC) and its surrounding atypical pericentriolar matrix form a dynamic basal complex (DBC) that facilitates a cascade of internal sliding deformations, coupling tail beating with asymmetric head kinking. During asymmetric tail beating, the DC's right side and its surroundings slide ~300 nm rostrally relative to the left side. The deformation throughout the DBC is transmitted to the head-tail junction; thus, the head tilts to the left, generating a kinking motion. These findings suggest that the DBC evolved as a dynamic linker coupling sperm head and tail into a single self-coordinated system.
Assuntos
Motilidade dos Espermatozoides/fisiologia , Animais , Centríolos/fisiologia , Centríolos/ultraestrutura , Humanos , Masculino , Mamíferos , Microtúbulos/fisiologia , Microtúbulos/ultraestrutura , Cabeça do Espermatozoide/fisiologia , Cauda do Espermatozoide/fisiologia , Cauda do Espermatozoide/ultraestruturaRESUMO
Tomcats are considered to be adults at 1 year of age, although many reach sexual maturity at an earlier age. Nevertheless, we still know little about whether the spermatogenic activity and sperm quality of mature under one-year-old tomcats differ from those of tomcats that are over one-year-old. This study aims to evaluate the spermatogenic activity, sperm traits, and their relationships in mature tomcats at two different ages. Sixteen tomcats showing complete spermatogenesis and spermatozoa in their epididymal caudae were used and classified according to their age as post-pubertal (<1 year old) or adult (Ë1 year old). Our results show that adult cats have higher epididymal sperm concentration and lower coefficient of variation in sperm head width and ellipticity than post-pubertal cats. However, they do not differ in their testicular and epididymal mass, spermatogenesis, and sperm traits such as motility, mitochondrial activity, morphology, morphometry, as well as plasma membrane, acrosome, and DNA integrity. Reduced intra-male variation of sperm head ellipticity is associated with higher testis mass, epididymis mass, and sperm concentration. Interestingly, low intra-male variation in sperm head size is associated with increased Sertoli cell function and reduced post-meiotic germ cell loss. These findings increase our knowledge about feline reproductive physiology and provide new insights into the functional significance of low intra-male variation in sperm size and shape in tomcats.
Assuntos
Variação Biológica da População , Tamanho Celular , Epididimo/crescimento & desenvolvimento , Cabeça do Espermatozoide/fisiologia , Espermatogênese , Testículo/crescimento & desenvolvimento , Fatores Etários , Animais , Gatos , Fertilidade , Masculino , Desenvolvimento Sexual , Contagem de Espermatozoides , Motilidade dos EspermatozoidesRESUMO
The beetle family Carabidae, with about 40,000 species, exhibits enough diversity in sperm structure and behavior to be an excellent model system for studying patterns and processes of evolution. We explore their potential, documenting sperm form in 177 species of ground beetles using light microscopy and collecting data on one qualitative and seven quantitative phenotypic traits. Our sampling captures 61% of the tribal-level diversity of ground beetles. These data highlight the notable morphological diversity of sperm in ground beetles and suggest that sperm in the group have dynamic evolutionary histories with much morphological innovation and convergence. Sperm vary among species in total length (48-3,400 µm), head length (0.5-270 µm), and head width (0.2-6.3 µm). Most ground beetles make sperm with heads that are indistinct from the flagella at the gross morphological level. However, some or all Omophron, Trachypachus, and Dyschiriini make broad-headed sperm that show morphological differences between species. Most ground beetles package their sperm into groups of sperm, termed conjugates, and ground beetles show variation in conjugate form and in the number and arrangement of sperm in a conjugate. Most ground beetles make sperm conjugates by embedding their sperm in a hyaline rod or spermatostyle. The spermatostyle is remarkably variable among species and varies in length from 17 to 41,000 µm. Several unrelated groups of ground beetles make only singleton sperm, including Nebriinae, Cicindelinae, many Trechinae, and the tribe Paussini. In order to study patterns in sperm evolution, we combine these data with a low-resolution phylogeny of ground beetles. Results from modern comparative analyses suggest the following: (a) sperm differ from conjugates in some aspect of their underlying evolutionary process, (b) sperm have influenced conjugate evolution and vice versa, and (c) conjugation with a spermatostyle likely evolved early within the history of Carabidae and it has been lost independently at least three times.
Assuntos
Evolução Biológica , Besouros/anatomia & histologia , Besouros/fisiologia , Espermatozoides/fisiologia , Análise de Variância , Animais , Feminino , Genitália Feminina/anatomia & histologia , Masculino , Modelos Biológicos , Fenótipo , Filogenia , Análise de Componente Principal , Característica Quantitativa Herdável , Cabeça do Espermatozoide/fisiologiaRESUMO
In cilia and flagella, dyneins form complexes which give rise to the inner and outer axonemal arms. Defects in the dynein arms are the leading cause of primary ciliary dyskinesia (PCD), which is characterized by chronic respiratory infections, situs inversus, and sterility. While the pathological features associated with PCD are increasingly well characterized, many of the causative genetic lesions remain elusive. Using Drosophila, here we analyze genetic requirements for wampa (wam), a previously uncharacterized component of the outer dynein arm. While homozygous mutant animals are viable and display no morphological defects, loss of wam results in complete male sterility. Ultrastructural analysis further reveals that wam mutant spermatids lack the axonemal outer dynein arms, which leads to a complete loss of flagellar motility. In addition to a role in outer dynein arm formation, we also uncover other novel microtubule-associated requirements for wam during spermatogenesis, including the regulation of mitochondrial localization and the shaping of the nuclear head. Due to the conserved nature of dyneins, this study advances our understanding of the pathology of PCD and the functional role of dyneins in axoneme formation and other aspects of spermatogenesis.
Assuntos
Axonema/metabolismo , Proteínas de Drosophila/metabolismo , Dineínas/metabolismo , Fertilidade/fisiologia , Cauda do Espermatozoide/metabolismo , Espermatogênese/fisiologia , Animais , Axonema/genética , Proteínas de Drosophila/genética , Drosophila melanogaster , Dineínas/genética , Infertilidade Masculina/genética , Infertilidade Masculina/metabolismo , Masculino , Cabeça do Espermatozoide/fisiologia , Motilidade dos Espermatozoides/fisiologiaRESUMO
Three genes are known to be essential for gamete adhesion/fusion (Cd9, Izumo1 and Juno). Here, we confirmed that Spaca6 null males are infertile and showed that their sperm accumulate in the perivitelline space but are unable to fuse with oocyte. Like IZUMO1, SPACA6 which is expressed by human sperm, is remained on the equatorial segment after acrosomal reaction and is involved in human fertilization since an anti-SPACA6 antibody inhibited it. Despite the similarity of the phenotypes caused by Spaca6 and Izumo1 knockouts, these are not redundant and the essential relocation of IZUMO1 is not affected by the lack of SPACA6. We propose a model in which IZUMO1 and SPACA6 would be part of a molecular complex necessary for gamete fusion and that their concomitant presence would be required for the recruitment of another essential molecular actor, such as a fusogen, for the fusion to take place.
Assuntos
Proteínas de Plasma Seminal/metabolismo , Interações Espermatozoide-Óvulo/fisiologia , Espermatozoides/fisiologia , Reação Acrossômica , Animais , Células COS , Chlorocebus aethiops , Feminino , Fertilização in vitro , Humanos , Imunoglobulinas/genética , Imunoglobulinas/metabolismo , Infertilidade Masculina/genética , Infertilidade Masculina/terapia , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Camundongos Knockout , Camundongos Transgênicos , Proteínas de Plasma Seminal/genética , Cabeça do Espermatozoide/fisiologia , Injeções de Esperma IntracitoplásmicasRESUMO
Saimiri collinsi is used as an animal model in biotechnology research for conservation of species from the genus Saimiri. However, the development of biotechnologies depends on a proper knowledge of the sperm morphology to understand the basic aspects of sperm physiology, as potential male fertility depends on different cellular sperm structures. With this purpose, this study characterized the micromorphological and ultrastructural characteristics of squirrel monkeys (Saimiri collinsi) sperm using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). SEM electromyography revealed that a normal Saimiri collinsi sperm measures 71.7 ± 0.7 µm with lateral tail insertion, a paddle-shaped flattened head and an acrosome occupying most of the head. TEM also showed that the middle piece is characterized by a central 9 + 2 microtubule axoneme surrounded by nine dense fibres, and that the mitochondria were juxtaposed, forming the mitochondrial sheath. Here we provide the first micromorphological and ultrastructure description of S. collinsi sperm.
Assuntos
Acrossomo/ultraestrutura , Cabeça do Espermatozoide/ultraestrutura , Cauda do Espermatozoide/ultraestrutura , Espermatozoides/ultraestrutura , Acrossomo/fisiologia , Animais , Axonema/ultraestrutura , Membrana Celular/ultraestrutura , Núcleo Celular/ultraestrutura , Humanos , Masculino , Microscopia Eletrônica de Varredura/métodos , Microscopia Eletrônica de Transmissão/métodos , Mitocôndrias/ultraestrutura , Sêmen/citologia , Cabeça do Espermatozoide/fisiologia , Motilidade dos Espermatozoides , Cauda do Espermatozoide/fisiologia , Espermatozoides/fisiologiaRESUMO
This study aimed to describe the morphology and sperm quality of free-living adult males of cururu stingray Potamotrygon wallacei, endemic from the Rio Negro basin, Brazilian Amazon. The sperm was collected in loco from the seminal vesicle region and fixed in buffered saline formaldehyde solution for further evaluation of morphometry, sperm plasma membrane integrity and sperm concentration. The spermatozoa presented a total length of 138.25 ± 1.82 µm with a helical shape and a long head. A high percentage of cells with intact membrane (98 ± 2%) and normal spermatozoa (92 ± 1%) were observed. The cell concentration was 0.34 ± 0.05 × 1010 spermatozoa/ml of semen. These observations are unprecedented for potamotrygonid species and will serve as a basis for future management and conservation strategies.
Assuntos
Reprodução/fisiologia , Sêmen/fisiologia , Rajidae/fisiologia , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/fisiologia , Animais , Brasil , Membrana Celular/fisiologia , Conservação dos Recursos Naturais/métodos , Água Doce , Geografia , Masculino , Rios , Sêmen/citologia , Cabeça do Espermatozoide/fisiologiaRESUMO
The development of analytical methods for the evaluation of crocodilian semen is an important component for the assessment of male breeding soundness and the development of assisted breeding technology in this taxon. Computer-Assisted Semen Analysis (CASA) technology is becoming an increasingly common technique in seminal evaluations for animals but there has been no application of this technique for reptilian spermatozoa. The aim of this study was to analyse sperm kinematic and morphometric variables in Caiman crocodilus fuscus semen samples and to determine whether there were sperm subpopulations. Four ejaculates from four sexually mature captive caimans were used for this study. A CASA-Mot and CASA-Morph system was used with an image acquisition rate of 50â¯Hz for 2â¯s of capture. The ISAS®D4C20 counting chambers were used and spermatozoa incubated at 25⯰C. Total and progressive motilities did not differ among animals (Pâ¯>â¯0.05). There was a significant animal effect in the model with respect to sperm morphometry, and kinematic indices including linearity (LIN) and straightness (STR) (Pâ¯<â¯0.05). Results for principal component (PC) analysis indicated variables were grouped into four components: PC1 related to velocity, PC2 to progressivity, PC3 to oscillation and PC4 to sperm path cross-linking. Subpopulation (SP) structure analysis indicated there were four groups, namely, rapid non-progressive (SP1), slow non-progressive (SP2), rapid progressive (SP3) and medium progressive (SP4), representing 14.5%, 45.4%, 18.7%, and 21.4% respectively. Findings in the present study indicate the importance of continuing development of reliable protocols regarding the standardisation of computer-based semen analyses in reptilian species.
Assuntos
Jacarés e Crocodilos , Análise do Sêmen/veterinária , Animais , Fenômenos Biomecânicos/fisiologia , Masculino , Reprodução/fisiologia , Cabeça do Espermatozoide/fisiologia , EspermatozoidesRESUMO
BACKGROUND: Infertility is a global health concern, and couples are increasingly seeking medical assistance to achieve reproduction. Semen analysis is a primary assessment performed by a clinician, in which the morphology of the sperm population is evaluated. Machine learning algorithms that automate, standardize, and expedite sperm classification are the subject of ongoing research. METHOD: We demonstrate a deep learning method to classify sperm into one of several World Health Organization (WHO) shape-based categories. Our method uses VGG16, a deep convolutional neural network (CNN) initially trained on ImageNet, a collection of human-annotated everyday images, which we retrain for sperm classification using two freely-available sperm head datasets (HuSHeM and SCIAN). RESULTS: Our deep learning approach classifies sperm at high accuracy and performs well in head-to-head comparisons with earlier approaches using identical datasets. We demonstrate improvement in true positive rate over a classifier approach based on a cascade ensemble of support vector machines (CE-SVM) and show similar true positive rates as compared to an adaptive patch-based dictionary learning (APDL) method. Retraining an off-the-shelf VGG16 network avoids excessive neural network computation or having to learn and use the massive dictionaries required for sparse representation, both of which can be computationally expensive. CONCLUSIONS: We show that our deep learning approach to sperm head classification represents a viable method to automate, standardize, and accelerate semen analysis. Our approach highlights the potential of artificial intelligence technologies to eventually exceed human experts in terms of accuracy, reliability, and throughput.
Assuntos
Aprendizado Profundo , Interpretação de Imagem Assistida por Computador/métodos , Análise do Sêmen/métodos , Espermatozoides/classificação , Algoritmos , Humanos , Masculino , Cabeça do Espermatozoide/classificação , Cabeça do Espermatozoide/fisiologia , Espermatozoides/fisiologiaRESUMO
Family with sequence similarity 46, member C (FAM46C) is a highly conserved non-canonical RNA polyadenylation polymerase that is abundantly expressed in human and mouse testes and is frequently mutated in patients with multiple myeloma. However, its physiological role remains largely unknown. In this study, we found that FAM46C is specifically localized to the manchette of spermatids in mouse testes, a transient microtubule-based structure mainly involved in nuclear shaping and intra-flagellar protein traffic. Gene knockout of FAM46C in mice resulted in male sterility, characterized by the production of headless spermatozoa in testes. Sperm heads were intermittently found in the epididymides of FAM46C knockout mice, but their fertilization ability was severely compromised based on the results of intracytoplasmic sperm injection assays. Interestingly, our RNA-sequencing analyses of FAM46C knockout testes revealed that mRNA levels of only nine genes were significantly altered compared to wild-type ones (q < 0.05). When considering alternate activities for FAM46C, in vitro assays demonstrated that FAM46C does not exhibit protein kinase or AMPylation activity against general substrates. Together, our data show that FAM46C in spermatids is a novel component in fastening the sperm head and flagellum.
Assuntos
Flagelos/fisiologia , Polinucleotídeo Adenililtransferase/fisiologia , Cabeça do Espermatozoide/fisiologia , Espermátides/fisiologia , Espermatogênese/genética , Animais , Diferenciação Celular/genética , Células Cultivadas , Feminino , Flagelos/metabolismo , Infertilidade Masculina/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Polinucleotídeo Adenililtransferase/genética , Gravidez , Cabeça do Espermatozoide/metabolismo , Espermátides/citologia , Espermatozoides/fisiologiaRESUMO
The postacrosomal sheath (PAS) of the perinuclear theca (PT) is the first compartment of the sperm head to solubilize into the ooplasm upon sperm-oocyte fusion, implicating its constituents in zygotic development. This study investigates the role of one such constituent, glutathione-S-transferase omega 2 (GSTO2), an oxidative-reductive enzyme found in the PAS and perforatorial regions of the PT. GSTO2 uses the conjugation of reduced glutathione, an electron donor shown to be compulsory in sperm disassembly within the ooplasm. The proximity of GSTO2 to the condensed sperm nucleus led us to hypothesize that this enzyme may facilitate nuclear decondensation by reducing disulfide bonds before the recruitment of GSTO enzymes from within the ooplasm. To test this hypothesis, we utilized a cell permeable isozyme-specific inhibitor, which fluoresces when bound to the active site of GSTO2, to functionally inhibit spermatozoa before performing intracytoplasmic sperm injections (ICSI) in mice. The technique allowed for targeted inhibition of solely PT-residing GSTO2, as all that is required for complete zygotic development is the injection of the mouse spermatozoon head. ICSI showed that inhibition of PT-anchored GSTO2 caused a delay in sperm nuclear decondensation, and further resulted in untimely embryo cleavage, and an increase in fragmentation beginning at the morula stage. The confounding effects of these developmental delays ultimately resulted in decreased blastocyst formation. This study implicates PT-anchored GSTO2 as an important facilitator of nuclear decondensation and reinforces the notion that the PAS-PT is a critical sperm compartment harboring molecules that facilitate zygotic development.
Assuntos
Glutationa Transferase/metabolismo , Cabeça do Espermatozoide/fisiologia , Espermatozoides/enzimologia , Sequência de Aminoácidos , Animais , Feminino , Glutationa Transferase/química , Glutationa Transferase/genética , Masculino , Camundongos , Injeções de Esperma Intracitoplásmicas/métodos , Interações Espermatozoide-Óvulo/fisiologiaRESUMO
BACKGROUND: Previous studies suggested that sperm head shape may serve as an effective indicator of semen quality. However, there lacks research with large sample and quantitative measurement. OBJECTIVES: The objective of this retrospective study was to explore the association between sperm head elongation (Width/Length ratio) and routine semen parameters. MATERIALS AND METHODS: From January 2012 to December 2017, 63 866 semen samples were collected from male subjects at 18-60 years of age. Sperm head elongation and routine semen parameters (semen volume, sperm concentration, motility, etc.) were examined with computer-assisted semen analysis (CASA) systems in order to evaluate the association between elongation and semen quality. RESULTS: Logistic and linear regression models showed that the value of elongation is negatively correlated with sperm concentration, total sperm count, progressive motility, total motility, percentage of morphologically normal spermatozoa, and acrosin activity (all p < 0.001). DISCUSSION: The results suggested that higher value of elongation is generally associated with higher risks of abnormality in semen quality. The importance of elongation may be explained by abnormal acrosin activity in the round-headed spermatozoa, which has been reported to cause failure of natural pregnancy. CONCLUSIONS: This study provides a new insight into the sperm head shape, which may be used as a complementary parameter in clinical semen examination and academic research.
Assuntos
Sêmen/fisiologia , Cabeça do Espermatozoide/fisiologia , Motilidade dos Espermatozoides/fisiologia , Teratozoospermia/fisiopatologia , Acrosina/metabolismo , Adolescente , Adulto , Humanos , Infertilidade Masculina , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Análise do Sêmen , Contagem de Espermatozoides , Adulto JovemRESUMO
Sperm morphology varies enormously across the animal kingdom. Whilst knowledge of the factors that drive the evolution of interspecific variation in sperm morphology is accumulating, we currently have little understanding of factors that may constrain evolutionary change in sperm traits. We investigated whether susceptibility to sperm abnormalities could represent such a constraint in songbirds, a group characterized by a distinctive helical sperm head shape. Specifically, using 36 songbird species and data from light and scanning electron microscopy, we examined among-species correlations between the occurrence of sperm head abnormalities and sperm morphology, as well as the correlation between sperm head abnormalities and two indicators of sperm competition. We found that species with more helically shaped sperm heads (i.e., a wider helical membrane and more pronounced cell waveform) had a higher percentage of abnormal sperm heads than species with less helical sperm (i.e., relatively straight sperm) and that sperm head traits were better predictors of head abnormalities than total sperm length. In contrast, there was no correlation between sperm abnormalities and the level of sperm competition. Given that songbird species with more pronounced helical sperm have higher average sperm swimming speed, our results suggest an evolutionary trade-off between sperm performance and the structural integrity of the sperm head. As such, susceptibility to morphological abnormalities may constrain the evolution of helical sperm morphology in songbirds.
Assuntos
Evolução Biológica , Aves Canoras/fisiologia , Cabeça do Espermatozoide/fisiologia , Animais , Masculino , Motilidade dos Espermatozoides/fisiologiaRESUMO
The binding of exogenous DNA to sperm is a key process for sperm-mediated gene transfer; however, the underlying molecular mechanisms have yet to be elucidated. In the present study, we aimed to identify the DNA binding proteins (DBPs) in rabbit sperm and to gain further understanding of the molecular mechanism of sperm and exogenous DNA interaction. Native polyacrylamide gel electrophoresis was used for separating free sperm proteins and complexes of DNA fragment/sperm proteins. A distinct band was found after Coomassie blue staining, and seven potential proteins were identified by mass spectrometry analysis. An analysis of the physical/chemical properties of the seven proteins revealed that the sperm inner acrosomal membrane protein IAM38 (IAM38) matched the features of the DBPs. Western blotting analysis showed that the IAM38 and CD4 were present in the sperm but not in the seminal plasma. Blocking of the IAM38 impaired the DNA-binding capacity of the sperm. Blocking the CD4 decreased the DNA-uptake capacity of the sperm but did not influence the DNA-binding capacity of the sperm. Moreover, the EGFP-positive embryos and EGFP-positive blastocysts were also decreased after IAM38 blocking or CD4 blocking in comparison with the control group. In conclusion, our results imply that foreign DNA first binds to the transmembrane IAM38 of the sperm plasma membrane and then forms the complex of DNA/IAM38/CD4 with CD4 to complete the transportation of exogenous DNA into the nucleus of sperm.
Assuntos
Acrossomo/metabolismo , Proteínas de Ligação a DNA/isolamento & purificação , Espermatozoides/metabolismo , Acrossomo/fisiologia , Animais , Blastocisto/metabolismo , Antígenos CD4/metabolismo , Membrana Celular/metabolismo , DNA/análise , Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação a DNA/fisiologia , Ensaio de Desvio de Mobilidade Eletroforética/métodos , Masculino , Coelhos , Cabeça do Espermatozoide/fisiologia , Espermatozoides/fisiologiaRESUMO
Sperm that swim collectively to the fertilization site have been observed across several vertebrate and invertebrate species, with groups ranging in size from sperm pairs to massive aggregates containing hundreds of cells. Although the molecular mechanisms that regulate sperm-sperm adhesion are still unclear, aggregation can enhance sperm motility and thus offer a fertilization advantage. Here, we report a thorough computational investigation on the role of cellular geometry in the performance of sperm aggregates. The sperm head is modelled as a persistent random walker characterized by a non-trivial three-dimensional shape and equipped with an adhesive region where cell-cell binding occurs. By considering both, a simple parametric head shape and a computer reconstruction of a real head shape based on morphometric data, we demonstrate that the geometry of the head and the structure of the adhesive region crucially affects both the stability and motility of the aggregates. Our analysis further suggests that the apical hook commonly found in the sperm of muroid rodents might serve to shield portions of the adhesive region and promote efficient alignment of the velocities of the interacting cells.
Assuntos
Forma Celular/fisiologia , Simulação por Computador , Modelos Biológicos , Cabeça do Espermatozoide/fisiologia , Motilidade dos Espermatozoides/fisiologia , Cauda do Espermatozoide/fisiologia , Animais , Masculino , RoedoresRESUMO
With the advent of sperm sex sorting methods and computer-aided sperm analysis platforms, comparative 2D motility studies showed that there is no significant difference in the swimming speeds of X-sorted and Y-sorted sperm cells, clarifying earlier misconceptions. However, other differences in their swimming dynamics might have been undetectable as conventional optical microscopes are limited in revealing the complete 3D motion of free-swimming sperm cells, due to poor depth resolution and the trade-off between field-of-view and spatial resolution. Using a dual-view on-chip holographic microscope, we acquired the full 3D locomotion of 235X-sorted and 289 Y-sorted bovine sperms, precisely revealing their 3D translational head motion and the angular velocity of their head spin as well as the 3D flagellar motion. Our results confirmed that various motility parameters remain similar between X- and Y-sorted sperm populations; however, we found out that there is a statistically significant difference in Y-sorted bovine sperms' preference for helix-shaped 3D swimming trajectories, also exhibiting an increased linearity compared to X-sorted sperms. Further research on e.g., the differences in the kinematic response of X-sorted and Y-sorted sperm cells to the surrounding chemicals and ions might shed more light on the origins of these results.
Assuntos
Imageamento Tridimensional , Análise para Determinação do Sexo , Cabeça do Espermatozoide/fisiologia , Motilidade dos Espermatozoides/fisiologia , Cauda do Espermatozoide/fisiologia , Espermatozoides/fisiologia , Animais , Bovinos , Feminino , MasculinoRESUMO
OBJECTIVE: Assess changes in sialic acid distribution during capacitation and acrosome reaction processes, and evaluate head sperm morphometrics modifications in these physiological conditions in human sperm. MATERIAL AND METHOD: In this prospective study, we included 6 normozoospermics sperm samples. Sialic acid distribution was evaluated by Wheat germ agglutinin lectin in different physiological conditions: before, after capacitation and after acrosome reaction. Head shape and size of each stage were analyzed by means of geometric morphometric methods. RESULTS: After capacitation, 73.07±21.43% of sperm showed sialic acid in acrosomal region, linked with an acrosome expansion and equatorial segment contraction. Otherwise, after acrosome reaction higher allometric effect between stages was recorded since sperm undergo further expansion of equatorial segment. Regarding Wheat germ agglutinin location, we found that sperm percentage significant decline in acrosomal fluorescence and an increase of equatorial band labeling. CONCLUSIONS: Our findings demonstrate that modifications in Wheat germ agglutinin expression covariate with dramatic changes in sperm head morphometry, suggesting important implications in capacitation and acrosome reaction processes.