RESUMO
The effect of inbreeding depression on sperm motility is well documented, but its influence on sperm morphometry has been scarcely examined to date. Here, we combined the use of computer-assisted sperm morphometry analysis (CASMA) with a SNP-based genomic approach to determine and characterize the effect of inbreeding on the sperm shape of a highly inbred cattle population. We determined seven morphometric parameters on frozen-thawed sperm samples of 57 Retinta bulls: length (L, µm), width (W, µm), area (A, µm2 ), perimeter (P, µm), ellipticity (ELI; L/W), elongation (L-W)/(L + W) and perimeter-to-area shape factor (p2a; P2 /4 × π × A). The comparison of highly inbred (HI) and lowly inbreed (LI) individuals based on runs of homozygosity (ROH) inbreeding values (F ROH ) showed no differences between groups. An additional two-step unsupervised sperm subpopulation analysis based on morphometric parameters showed significant differences in the abundance of different sperm subpopulations between groups (p < 0.05). This analysis revealed that HI bulls harbored a higher percentage of narrow-head sperm as opposed to the higher percentage of large- and round-headed sperm detected in LI. A further genomic characterization revealed 23 regions differentially affected by inbreeding in both groups, detecting six genes (SPAG6, ARMC3, PARK7, VAMP3, DYNLRB2, and PHF7) previously related to different spermatogenesis-associated processes.
Assuntos
Bovinos/genética , Depressão por Endogamia/genética , Endogamia , Espermatozoides/ultraestrutura , Animais , Animais Endogâmicos , Variação Biológica Individual , Forma Celular , DNA/genética , Estudos de Associação Genética , Genótipo , Haplótipos/genética , Masculino , Cabeça do Espermatozoide/ultraestruturaRESUMO
The objective of this study was to describe and compare the head morphometry of normal and pathological sperm from Saimiri macrodon. In the morphological analysis, 39% of the sperm had normal morphology, and 61% had major or minor defects, with pathology in the tails being the most frequent with 47% (38% curled tail, 7% folded tail and 2% strongly folded tail). Among the evaluated head morphometry parameters, area (A), width (L) and ellipticity (E) showed statistical difference (p>0.05) between normal and pathological sperm. The average head area and width was lower in normal sperm (p=0,01 e p=0,04, respectively), and the mean ellipticity was higher (p=0,038), when compared to pathological sperm. This definition of the sperm morphometric parameters of S. macrodon is important for the samples selection destined to reproduction biotechnologies and for the clarification of taxonomic and evolutionary issues in the genus Saimiri.
Assuntos
Animais , Masculino , Saimiri/anatomia & histologia , Cabeça do Espermatozoide/ultraestrutura , Biometria , Animais Selvagens/anatomia & histologiaRESUMO
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
In Northern Patagonia, the mating season starts on March 15th, when rams are submitted to summer temperatures. Exposure of rams to heat stress increases the prevalence of microscopic damage to spermatozoa, morphological abnormalities, and reductions in fertility. This study assesses the adaptive capabilities of six unshorn and six shorn Australian Merino rams, half of which were treated in a heat chamber for eight hours for five days, gradually reaching a temperature of up to 40 °C. Microscopic damage, abnormalities and ultramicroscopic alterations of the plasma membrane and the acrosome of sperm head were analysed. There were significant differences in the percentage of tailless spermatozoa and proximal cytoplasmic droplets between post-treatment periods. Temperature primarily affected the shorn rams and the sperm heads during spermiogenesis. Submicroscopic alterations were observed when the plasma membrane was present in the anterior segment. These alterations can be intact, waved, or dilated. When the plasma membrane was absent, the acrosome might be intact, dilated, and waved. In addition, the outer acrosomal membrane may completely lose its contents or have a nude nucleus. The plasma membrane assumes a waved shape as a result of the effect of temperature on the epididymis. According to this study, the tailless head, proximal cytoplasmic droplets, and the ultramicroscopic categories studied were robust indicators of semen heat stress. After ten weeks, the sperm head recovered its normal shape. Unshorn rams are better adapted to summer heat stress than shorn ones. Microscopy and transmission electron microscopy alterations have been shown to be excellent indicators of thermal stress in Australian Merino rams and may be useful tools to help sheep farmers choose when to begin the mating season, which will vary depending on the environmental conditions of the summer.(AU)
Na Patagônia Norte, os ovinos têm sua estação de acasalamento iniciada em 15 de março, portanto, ficam sujeitos às temperaturas do verão. A exposição de carneiros a estresse térmico aumenta a prevalência de danos microscópicos e anomalias morfológicas nos espermatozoides, que implica uma redução na fertilidade. Este trabalho avaliou a capacidade adaptativa de carneiros Merino Australiano com lã (N = 6) e tosquiados (N = 6): metade ficou ao ar livre e outra metade foi mantida em uma câmara climática por oito horas, durante cinco dias, chegando gradualmente a uma temperatura máxima de 40 °C. Foram analisados danos microscópicos, anormalidades e alterações ultramicroscópicas da membrana plasmática e do acrossoma da cabeça dos espermatozoides. Os resultados microscópicos confirmaram a existência de diferença significativa na porcentagem de espermatozoides sem cauda e com gota citoplasmática proximal, entre os ejaculados pós-tratamento. A temperatura afetou os carneiros tosquiados, principalmente a cabeça de seus espermatozoides, durante a espermatogênese. Alterações submicroscópicas foram observados na membrana plasmática quando ela estava presente no segmento anterior: quando não intacta, ficava ondulada ou dilatada. Quando a membrana plasmática estava ausente, o acrossoma podia se apresentar ondulado ou dilatado. Além disso, sob efeito do calor, a membrana acrossomal externa pode perder completamente seu conteúdo ou apresentar núcleo desnudo. A membrana plasmática assume uma forma ondulada pelo efeito da temperatura no epidídimo. Depois de dez semanas, a cabeça dos espermatozoides recuperou sua forma normal. Como demonstrado neste estudo, a cabeça sem cauda, as gotas citoplasmáticas proximais e as categorias ultramicroscópicas estudadas são indicadores do efeito do estresse térmico no sêmen, e os carneiros com maior cobertura de lã se adaptam melhor ao estresse por calor. Alterações de microscopia e de microscopia eletrônica de transmissão têm se mostrado excelentes indicadores de estresse por calor em carneiros Merino Australiano e podem ser ferramentas úteis para ajudar criadores de ovelhas a escolher quando começar a época de acasalamento, o que irá variar de acordo com as condições ambientais do verão.(AU)
Assuntos
Animais , Masculino , Acrossomo/ultraestrutura , Membrana Celular/ultraestrutura , Transtornos de Estresse por Calor/complicações , Ovinos/fisiologia , Cabeça do Espermatozoide/ultraestrutura , Teratozoospermia/diagnóstico por imagem , Argentina , Cauda do Espermatozoide/ultraestrutura , EspermatogêneseRESUMO
In Northern Patagonia, the mating season starts on March 15th, when rams are submitted to summer temperatures. Exposure of rams to heat stress increases the prevalence of microscopic damage to spermatozoa, morphological abnormalities, and reductions in fertility. This study assesses the adaptive capabilities of six unshorn and six shorn Australian Merino rams, half of which were treated in a heat chamber for eight hours for five days, gradually reaching a temperature of up to 40 °C. Microscopic damage, abnormalities and ultramicroscopic alterations of the plasma membrane and the acrosome of sperm head were analysed. There were significant differences in the percentage of tailless spermatozoa and proximal cytoplasmic droplets between post-treatment periods. Temperature primarily affected the shorn rams and the sperm heads during spermiogenesis. Submicroscopic alterations were observed when the plasma membrane was present in the anterior segment. These alterations can be intact, waved, or dilated. When the plasma membrane was absent, the acrosome might be intact, dilated, and waved. In addition, the outer acrosomal membrane may completely lose its contents or have a nude nucleus. The plasma membrane assumes a waved shape as a result of the effect of temperature on the epididymis. According to this study, the tailless head, proximal cytoplasmic droplets, and the ultramicroscopic categories studied were robust indicators of semen heat stress. After ten weeks, the sperm head recovered its normal shape. Unshorn rams are better adapted to summer heat stress than shorn ones. Microscopy and transmission electron microscopy alterations have been shown to be excellent indicators of thermal stress in Australian Merino rams and may be useful tools to help sheep farmers choose when to begin the mating season, which will vary depending on the environmental conditions of the summer.(AU)
Na Patagônia Norte, os ovinos têm sua estação de acasalamento iniciada em 15 de março, portanto, ficam sujeitos às temperaturas do verão. A exposição de carneiros a estresse térmico aumenta a prevalência de danos microscópicos e anomalias morfológicas nos espermatozoides, que implica uma redução na fertilidade. Este trabalho avaliou a capacidade adaptativa de carneiros Merino Australiano com lã (N = 6) e tosquiados (N = 6): metade ficou ao ar livre e outra metade foi mantida em uma câmara climática por oito horas, durante cinco dias, chegando gradualmente a uma temperatura máxima de 40 °C. Foram analisados danos microscópicos, anormalidades e alterações ultramicroscópicas da membrana plasmática e do acrossoma da cabeça dos espermatozoides. Os resultados microscópicos confirmaram a existência de diferença significativa na porcentagem de espermatozoides sem cauda e com gota citoplasmática proximal, entre os ejaculados pós-tratamento. A temperatura afetou os carneiros tosquiados, principalmente a cabeça de seus espermatozoides, durante a espermatogênese. Alterações submicroscópicas foram observados na membrana plasmática quando ela estava presente no segmento anterior: quando não intacta, ficava ondulada ou dilatada. Quando a membrana plasmática estava ausente, o acrossoma podia se apresentar ondulado ou dilatado. Além disso, sob efeito do calor, a membrana acrossomal externa pode perder completamente seu conteúdo ou apresentar núcleo desnudo. A membrana plasmática assume uma forma ondulada pelo efeito da temperatura no epidídimo. Depois de dez semanas, a cabeça dos espermatozoides recuperou sua forma normal. Como demonstrado neste estudo, a cabeça sem cauda, as gotas citoplasmáticas proximais e as categorias ultramicroscópicas estudadas são indicadores do efeito do estresse térmico no sêmen, e os carneiros com maior cobertura de lã se adaptam melhor ao estresse por calor. Alterações de microscopia e de microscopia eletrônica de transmissão têm se mostrado excelentes indicadores de estresse por calor em carneiros Merino Australiano e podem ser ferramentas úteis para ajudar criadores de ovelhas a escolher quando começar a época de acasalamento, o que irá variar de acordo com as condições ambientais do verão.(AU)
Assuntos
Animais , Masculino , Cabeça do Espermatozoide/ultraestrutura , Acrossomo/ultraestrutura , Ovinos/fisiologia , Membrana Celular/ultraestrutura , Transtornos de Estresse por Calor/complicações , Teratozoospermia/diagnóstico por imagem , Argentina , Cauda do Espermatozoide/ultraestrutura , EspermatogêneseRESUMO
The sperm morphology of two predatory bugs Amphiareus constrictus and Blaptostethus pallescens, representing the tribes Dufouriellini and Blasptostethini, respectively, was described using light and transmission electron microscopy. The spermatozoa of Amphiareus constrictus and Blaptostethus pallescens are fine and long, each measuring 216.6µm and 181.0µm in length, of which 37.0µm and 11.6µm, respectively, comprise the nuclei. When stained with DAPI (for DNA), the posterior half of the nucleus in B. pallescens exhibited low fluorescence, while in A. constrictus this feature was observed only in the last 6µm. In both species, as in Heteroptera in general, the spermatozoa have, in the head region, an acrosome and nucleus, and in the flagellar region, an axoneme with 9 accessory tubules, 9 peripheral doublets and 2 central microtubules (9+9+2 microtubules), 2 mitochondrial derivatives (MDs), and a centriolar adjunct in the nucleus-flagellum transition. However, unlike most Heteroptera, in these species, the MDs are asymmetric, and the centriolar adjunct is quite long and encompasses completely the posterior nuclear end and the anterior tips of the MDs. These features are considered as derived, thus supporting the condition derived of Anthocoridae within Cimicomorpha. In addition, several traits of the spermatozoa of these two species easily distinguish one species (and probably a tribe) from the other; for example, the difference of formats in the MDs, and the long anterior projection of the centriolar adjunct parallel to the nucleus in B. pallescens.
Assuntos
Heterópteros/anatomia & histologia , Heterópteros/ultraestrutura , Cabeça do Espermatozoide/ultraestrutura , Animais , Núcleo Celular/ultraestrutura , Centríolos/ultraestrutura , Masculino , Microscopia Eletrônica de Transmissão , Mitocôndrias/fisiologiaRESUMO
The aim was to study the ultrastructure of testicular parenchyma and define the morphological ultrastructure of spermatozoa of agoutis kept in captivity. Segments of testes from eight agouti males at prepubescence, prepuberty, pubescence and sexual maturity were fixed in glutaraldehyde. Laboratory procedures were performed for transmission electron microscopy. Spermatogonial cells of Type A - pale, Type A - dark, intermediate and Type B were found. Spermatocytes in the pachytene phase were abundant among primary spermatocytes. From the prepubertal phase, Sertoli cells exhibited invaginations in the nuclear membrane and lipid inclusions in the cytoplasm due to their phagocytic function. Leydig cells displayed higher metabolic activity during puberty as evidenced by the presence of lipid droplets. Spermatozoa were fully formed morphologically at prepuberty. The centriolar complex had partially degenerated and featured a centriolar space as in rodents. Sperm heads were tapered, without prominence of the acrosome or evidence of the perforatorium, differing from cavies, rats and mice. This is the first study to describe the ultrastructure of agouti spermatozoa. This research may assist as a basis for future work related to fertility and other biotechnologies applied to reproductive biology in agoutis.
Assuntos
Maturidade Sexual/fisiologia , Cabeça do Espermatozoide/ultraestrutura , Espermatogênese/fisiologia , Espermatozoides/ultraestrutura , Testículo/ultraestrutura , Animais , Dasyproctidae , Células Intersticiais do Testículo/ultraestrutura , Masculino , Células de Sertoli/ultraestrutura , Espermátides/ultraestrutura , Espermatócitos/ultraestrutura , Espermatogônias/ultraestruturaRESUMO
Semen analysis is the first step in the evaluation of an infertile couple. Within this process, an accurate and objective morphological analysis becomes more critical as it is based on the correct detection and segmentation of human sperm components. In this paper, we present an improved two-stage framework for detection and segmentation of human sperm head characteristics (including acrosome and nucleus) that uses three different color spaces. The first stage detects regions of interest that define sperm heads, using k-means, then candidate heads are refined using mathematical morphology. In the second stage, we work on each region of interest to segment accurately the sperm head as well as nucleus and acrosome, using clustering and histogram statistical analysis techniques. Our proposal is also characterized by being fully automatic, where a user intervention is not required. Our experimental evaluation shows that our proposed method outperforms the state-of-the-art. This is supported by the results of different evaluation metrics. In addition, we propose a gold-standard built with the cooperation of a referent expert in the field, aiming to compare methods for detecting and segmenting sperm cells. Our results achieve notable improvement getting above 98% in the sperm head detection process at the expense of having significantly fewer false positives obtained by the state-of-the-art method. Our results also show an accurate head, acrosome and nucleus segmentation achieving over 80% overlapping against hand-segmented gold-standard. Our method achieves higher Dice coefficient, lower Hausdorff distance and less dispersion with respect to the results achieved by the state-of-the-art method.
Assuntos
Aumento da Imagem/métodos , Interpretação de Imagem Assistida por Computador/normas , Microscopia/métodos , Microscopia/normas , Reconhecimento Automatizado de Padrão/normas , Análise do Sêmen/normas , Cabeça do Espermatozoide/ultraestrutura , Inteligência Artificial , Células Cultivadas , Chile , Humanos , Interpretação de Imagem Assistida por Computador/métodos , Masculino , Reconhecimento Automatizado de Padrão/métodos , Padrões de Referência , Reprodutibilidade dos Testes , Análise do Sêmen/métodos , Sensibilidade e EspecificidadeRESUMO
Fish belonging to the family Rivulidae possess one of the most complex reproductive systems. Rivulus, a genus of freshwater fish in the Rivulidae family, was recently reclassified into five genera, including Melanorivulus. Its type species, M. punctatus, is widely distributed and probably represents a species complex. The ultrastructure of sperm has been broadly used in systematics, and we hereby describe the ultrastructural features of spermatogenesis in M. punctatus. Ten M. punctatus males were collected from the reservoir of Parque Estadual da Quineira, municipality of Chapada dos Guimarães, Mato Grosso, Brazil, and prepared for analysis by light microscopy and transmission electron microscopy. M. punctatus undergoes cystic spermatogenesis. Its cysts consist of groups of germ cells that are in synchronous development and are surrounded by cytoplasmic projections of Sertoli cells. With the breakdown of the cysts, the spermatozoa are released and their maturation is completed in the duct, where part of the cytoplasmic material is discarded through the vesicles. The mature spermatozoon is characterized by a spherical head with homogeneously condensed chromatin, a symmetric midpiece consisting of a pair of perpendicular centrioles, a ring of mitochondria, several vesicles, and one flagellum medial to the nucleus. Early stages of spermatogenesis show no peculiarities; however, in spermiogenesis, we observed that the spermatids remain interconnected by cytoplasmic bridges and have pockets of residual cytoplasm. The sperm is of the aquasperm type and is similar to that observed in the members of the family Rivulidae. The spermatozoa have a single flagellum that consists of a classic axoneme (9+2), as found in most groups of fish, despite the lateral extensions.
Assuntos
Ciprinodontiformes/anatomia & histologia , Espermatogênese , Espermatozoides/ultraestrutura , Animais , Brasil , Masculino , Microscopia Eletrônica de Transmissão , Células de Sertoli/ultraestrutura , Cabeça do Espermatozoide/ultraestrutura , Cauda do Espermatozoide/ultraestrutura , Espermátides/ultraestruturaRESUMO
Sperm dimensions and the question of whether X and Y chromosome-bearing sperm differ in size or shape has been of great interest, especially for the development of alternative methods to sort or classify sperm cells. The aim of the present study was to evaluate possible differences in the shape and size of the sperm head between X and Y chromosome-bearing sperm by atomic force microscopy (AFM). One ejaculate per bull (nâ=â4) was used. Each ejaculate was separated into four fractions: non-sexed (NS), sexed for X-sperm (SX), sexed for Y-sperm (SY) and a pooling of SX and SY samples (SXY). Using AFM, 400 sperm heads per group were measured. Twenty three structural features were assessed including one-, two- and three-dimensional parameters and shape descriptors. These measurements determine the micro- to nanoscale features of X- and Y-bearing chromosomes in sperm cells. No differences were observed for any individual variables between SX and SY groups. Next, a simultaneous evaluation of all features using statistical discriminant analysis was performed to determine if it was possible to distinguish to which group belong each individual cells. This analysis clearly showed, a distinct separation of NS, SXY, SX and SY groups. The recognition of this structural possibility to distinguish between X and Y sperm cell might improve the understanding of sperm cells biology. These results indicated that the associations of several structural measurements of the sperm cell head are promising candidates for development of a new method of sperm sexing.
Assuntos
Forma Celular/fisiologia , Tamanho Celular , Microscopia de Força Atômica/veterinária , Análise para Determinação do Sexo/métodos , Cabeça do Espermatozoide/ultraestrutura , Cromossomo X/química , Cromossomo Y/química , Animais , Bovinos , Separação Celular/métodos , Separação Celular/veterinária , Análise Discriminante , Modelos Lineares , Masculino , Microscopia de Força Atômica/métodos , Cabeça do Espermatozoide/químicaRESUMO
The aim of this study was to evaluate the incidence of pleiomorphisms and its influence on the distribution of sperm morphometric subpopulations in ejaculates from the vulnerable Goeldi's monkey (Callimico goeldii) by using a combination of computerized analysis system and Principal Component Analysis (PCA) methods. Each sperm head was measured for four primary spermatozoal head dimensional parameters (area [A (µm(2))], perimeter [P (µm)], length [L (µm)] and width [W (µm)]) and three head shape derived parameters (ellipticity [(L/W)], elongation [(L-W)/(L+W)] and rugosity [(4πA/P(2))]). Six separate subpopulations (SPs) were identified: SP1, constituted by very large, narrow and very elliptical spermatozoa (A=16.85±1.56µm(2), W=2.75±0.42µm and ellipticity=2.16±0.24); SP2, characterized by average sized, short, wide and round spermatozoa (A=15.00±1.92µm(2), L=5.06±0.49µm, W=3.51±0.31µm and ellipticity=1.44±0.15); SP3, represented by small, wide and slightly round spermatozoa (A=14.95±1.75µm(2), W=3.47±0.29µm and ellipticity=1.48±0.14); SP4 included very small, short and very round spermatozoa (A=14.15±2.38µm(2), L=4.90±0.57µm and elongation=0.18±0.05); SP5 consisted of average sized and slightly elliptical spermatozoa (A=15.14±1.72µm(2) and ellipticity=1.49±0.14); and SP6 included large and round spermatozoa (A=16.30±1.62µm(2) and elongation=0.19±0.04). There were differences in the sperm subpopulation distribution (P<0.001) among the five donors analyzed. In conclusion, the results of the current study confirmed that the use of computer sperm analysis methods combined with PCA cluster analyses are useful methods to identify, classify, and characterize different sperm head morphometric subpopulations in neotropical primates. Broadening our knowledge of C. goeldii sperm morphometric abnormalities as well as developing reliable techniques for sperm evaluation may be essential for ex situ conservation of this threatened species.
Assuntos
Callimico/anatomia & histologia , Cabeça do Espermatozoide/ultraestrutura , Animais , Conservação dos Recursos Naturais , Processamento de Imagem Assistida por Computador , Masculino , Microscopia de Contraste de Fase/veterinária , Análise de Componente PrincipalRESUMO
In this study, the morphology of spermatozoa of Bethylidae Dissomphalus connubialis (Pristocerinae) was analyzed using light and transmission electron microscopy. Spermatozoa of this species are thin, measure approximately 130 µm in length and comprise a head region and flagellum region. The head is formed by the acrosome and nucleus. The acrosome consists of the acrosome vesicle and the perforatorium, the posterior portion of which is inserted into a cavity at the anterior extremity of the nucleus. The nucleus is compact, electron-dense and measures 15 µm in length. The flagellum possesses two mitochondrial derivatives, two accessory bodies and one axoneme with a 9+9+2 microtubular pattern. The nucleus is connected to the flagellum by the centriole adjunct. Mitochondrial derivatives are compact, apparently without paracrystalline material and with rare mitochondrial cristae. They are asymmetric in length, such that the larger mitochondrial derivative begins parallel to the posterior region of the nucleus and the smaller mitochondrial derivative begins just below the centriole adjunct. The basic structure of spermatozoa of D. connubialis is similar to that of other Aculeata studied. However, this species shows characteristics not seen in other Hymenoptera, such as the wide electron-lucid region that separates the acrosomal vesicle from the perforatorium and the depth of the cavity in the anterior extremity of the nucleus, into which the base of the perforatorium is inserted. There are also characteristics that distinguish this species from Bethylidae Prorops nasuta, including the fact that one of the mitochondrial derivatives lies to parallel to the nucleus over a long distance, the small quantity of cristae, the absence of paracrystalline material in these organelles, and the fact that the accessory microtubules are the first to terminate in the final portion of the flagellum.
Assuntos
Himenópteros/anatomia & histologia , Espermatozoides/ultraestrutura , Acrossomo/ultraestrutura , Animais , Núcleo Celular/ultraestrutura , Masculino , Microscopia Eletrônica de Transmissão , Mitocôndrias/ultraestrutura , Cabeça do Espermatozoide/ultraestrutura , Cauda do Espermatozoide/ultraestruturaRESUMO
Examination of the type and frequency of damage to the head of spermatozoa using electron microscopy can be used to evaluate the quality of differently treated sperm. This report describes a systematic approach based on 29 morphological categories of sperm heads assessed from discrete regions in raw, chilled and frozen-thawed spermatozoa. Injury occurred principally at the plasma membrane and could be present or absent in all regions. In the anterior segment, when the plasma membrane is present, it can be intact, dilated, very dilated, disrupted, or contain vesicles characteristic of acrosomal reaction-like capacitation changes. When the plasma membrane is absent, the acrosome may be intact, exhibit a complete loss of contents, or retain some contents of the apical ridge and present a very dilated outer acrosomal membrane. The plasma membrane in the equatorial segment and the boundary between regions can be intact, dilated, very dilated or disrupted. The post-acrosomal plasma membrane is classified as intact, dilated or very dilated, whereas the dense lamina is intact, dilated or fragmented. The morphology of the heads most frequently observed in chilled spermatozoa consists of anterior and equatorial segments with a dilated, or dilated and disrupted plasma membrane; a boundary between regions with an intact and dilated plasma membrane; and a post-acrosomal region with an intact plasma membrane and dense lamina, both dilated. In frozen-thawed spermatozoa, the morphology of the heads is more frequently characterised by no plasma membrane and an acrosome showing complete or some loss of contents in the apical ridge and very dilated outer acrosomal membrane, presenting mostly dilated and fragmented dense lamina in the post-acrosomal region. These findings are consistent with the conclusion that the freezing process produces an increase in the degree of damage to the cells when they are subjected to increasing degrees of cold shock. There are still difficulties in developing a good diluent and process for preserving the plasma membrane in ram spermatozoa. This systematisation, using different categories, allows characterisation of multiple transmission electron microscopy images. Thus, the different changes observed due to cryopreservation may be correlated.
Assuntos
Membrana Celular/ultraestrutura , Criopreservação , Cabeça do Espermatozoide/ultraestrutura , Animais , Membrana Celular/efeitos dos fármacos , Crioprotetores/farmacologia , Congelamento , Processamento de Imagem Assistida por Computador , Masculino , Microscopia Eletrônica de Transmissão , Carneiro Doméstico , Cabeça do Espermatozoide/efeitos dos fármacosRESUMO
Llama production in Argentina has increased, as the international interest in breeding this type of animals has grown in the last years. Considering the great polymorphism that llama spermatozoa present at evaluation using light microscopy, the aim of this study was to objectively evaluate llama sperm head morphometry using digital morphometric analysis. Five ejaculates from each of eight males were obtained to evaluate morphometric parameters of 8000 sperm heads stained with Tinción 15(®). The following average results were obtained for each parameter: size parameters: area 20.09 µm(2), length 6.60 µm, width 4.14 µm, equivalent circle diameter 5.06 µm, curve length 5.79 µm and curve width 3.48 µm; boundary parameters: perimeter 18.54 µm and convex perimeter 17.34 µm; and shape parameters: roundness 1.28 and elongation 1.59. Morphometric parameters of sperm head were compared between ejaculates of the same male and between males. Significant differences between ejaculates of the same male were found for all parameters evaluated (P < 0.01). Significant differences between males were found for all morphometric parameters (P < 0.01) except for curve length, curve width and perimeter. The differences detected would indicate that there is not a single morphometric pattern for Lama glama sperm head, because parameter values cannot be standardised.
Assuntos
Camelídeos Americanos , Cabeça do Espermatozoide/ultraestrutura , Animais , MasculinoRESUMO
The perinuclear theca (PT) is a cytoskeletal structure that surrounds the mammal sperm nucleus which must be disrupted once the sperm has penetrated the oocyte to permit normal chromatin decondensation and formation of male pronucleus. F-actin is a thermo sensitive protein found in the equatorial segment which is involved in the stability of PT. It has been reported that cryopreservation induces alterations in nuclear decondensation of spermatozoa, which have been interpreted as an over condensation. The aims of the present study were identified the presence of changes in sperm sPT integrity of frozen-thawed boar spermatozoa and its effect in sperm nuclei decondensation; and whether changes in the actin cytoskeleton are involved using an in vitro model to test probably differences in a chemical decondensation (DTT/heparin) between fresh (FS) and frozen-thawed (TS) spermatozoa. Results showed an increase on sPT damage in TS (P<0.001), and significant changes in sperm chromatin nuclear decondensation (P<0.05). In same way differences on the swelling degree was found assessed by measures in equatorial region of head sperm (P<0.05). Evaluation with rodamine-labeled actin (0.2µM) showed two different patterns with differences in percentages before and after cryopreservation (P<0.001). F-actin stabilization constrained the equatorial segment of FS while this was not observed in TS. The data showed that the presence of early changes in sPT integrity and changes in the F-actin localization on TS may suggest the participation in F-actin in decondensation process and probably that the disruption of actin-PT interaction during freezing-thawing process could have far-reaching consequences for the subsequent fertility of spermatozoa.
Assuntos
Actinas/metabolismo , Cromatina/ultraestrutura , Criopreservação/métodos , Congelamento/efeitos adversos , Preservação do Sêmen/efeitos adversos , Cabeça do Espermatozoide/ultraestrutura , Sus scrofa , Animais , Citoesqueleto/fisiologia , Tomografia com Microscopia Eletrônica/métodos , Masculino , Sêmen/citologia , Sêmen/fisiologia , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Cabeça do Espermatozoide/fisiologia , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/fisiologia , Espermatozoides/ultraestruturaRESUMO
A specific cause of infertility cannot be identified in at least 25% of men referred to a specialized clinic. Diagnosis of infertile men is based mainly on standard semen analysis and the observation of sperm under light microscope. The aim of our study was to find the subcellular sperm characteristics that could explain infertility in a group of teratozoospermic infertile men. Morphological characteristics of sperm from non-teratozoospermic (control donors) and teratozoospermic infertile men were analyzed by transmission electron microscopy (TEM) and quantified. Our analysis showed that sperm cells from control donors presented a higher number of normal heads and tails than infertile men. Regarding subcellular characteristics of nucleus and tails, only the percentage of vacuolated nucleus, the absence of at least one pair of microtubules of the axoneme and the total distortion of the tail were statistically higher in infertile men than in control donors. There were no differences in the number of normal acrosomes between the groups. Although the ultrastructural sperm defects overlapped between control donors and infertile men, TEM permits the identification and differentiation of a larger amount of defects than light microscopy. Vacuolated nucleus and gross alterations of the tail are the major sperm defects that seem to have prognostic value in teratozoospermic men.
Assuntos
Infertilidade Masculina/fisiopatologia , Espermatozoides/anormalidades , Espermatozoides/ultraestrutura , Adulto , Feminino , Humanos , Infertilidade Masculina/diagnóstico , Infertilidade Masculina/etiologia , Infertilidade Masculina/patologia , Masculino , Gravidez , Sêmen/citologia , Cabeça do Espermatozoide/patologia , Cabeça do Espermatozoide/ultraestrutura , Injeções de Esperma Intracitoplásmicas , Cauda do Espermatozoide/patologia , Cauda do Espermatozoide/ultraestrutura , Adulto JovemRESUMO
Fertilization in mammals occurs via a series of well-defined events in the secluded environment of the female reproductive tract. The mode of selection of the fertilizing spermatozoon nevertheless remains unknown. As has become evident during in vitro fertilization by sperm microinjection into the oocyte, abnormal spermatozoa can successfully fertilize oocytes. Under these extreme conditions, post-fertilization events, early embryonic development and implantation are significantly compromised indicating that the contribution of spermatozoa extends beyond sperm penetration. Microscopic identification of normal spermatozoa is a well-standardized procedure but insights into the mechanisms that lead to aberrant sperm differentiation and into the subcellular nature of sperm abnormalities have only recently begun to be obtained. The spermatozoon is the result of a complex development in which spermatid organelles give rise to various structural components with characteristic functions. Similar to other differentiated cells, the spermatozoon has a specific pathology that is most clearly identified by ultrastructural evaluation coupled with immunocytochemistry and molecular techniques. This multidisciplinary approach allows the precise characterization of sperm abnormalities, including structural, molecular and functional aspects. We summarize here studies of the physiopathology of spermiogenesis in two abnormal sperm phenotypes of infertile men: dysplasia of the fibrous sheath and acephalic spermatozoa/abnormal head-tail attachment. The characterization of the abnormalities of the tail cytoskeleton and centrioles has uncovered aspects of the subcellular basis of pathological spermiogenesis, has suggested experimental approaches to explore the nature of these anomalies and has opened the way for genetic studies that will ultimately lead to the design of the therapeutic tools of the future.
Assuntos
Espermatogênese/fisiologia , Espermatozoides/anormalidades , Espermatozoides/patologia , Animais , Feminino , Humanos , Infertilidade Masculina/etiologia , Infertilidade Masculina/patologia , Masculino , Cabeça do Espermatozoide/patologia , Cabeça do Espermatozoide/ultraestrutura , Espermatozoides/citologia , Espermatozoides/ultraestruturaRESUMO
The objectives of this study were to compare two different methods of evaluating bull sperm morphology, bright-field (BF) microscopy of eosin-nigrosin (EN) stained dry-mount semen smears and differential interference phase contrast (DIC) microscopy of wet-mount semen 'fixed' in isotonic formal saline, both at 1000x. Ejaculates (n=72) were evaluated, representing both pre- and post-breeding season ejaculates collected from 40 2-yr-old beef bulls via electro-ejaculation. For both methods, 200 sperm were counted in random fields with defects categorized as major (MAD) and minor (MID). Sperm abnormalities were also placed into two other categories: those considered to be most influenced by process (wet or dry, METHDEF) and those with depictions that could be influenced by optics (BF or DIC, OPTIDEF). Differences (P<0.05) occurred between DIC and BF methods respectively: MAD 23.3/16.1, MID 7.6/13.4, acrosome 3.8/1.1, midpiece 9.2/11.7, tail 2.0/4.7, droplets 8.3/4.2, METHDEF 14.2/21.4 and OPTIDEF 13.0/5.5, but not (P>0.05) in percent normal sperm 69.1/70.4 or sperm head defects 7.5/8.3. Acrosome, tail and droplet defects were observed in 98.2/80.5, 86.1/100 and 98.2/94.4 percent of bulls for DIC and BF, respectively (P<0.05). As percent normal sperm did not differ between methods, bright-field microscopy assessment of EN preparations was considered to be a satisfactory method to categorize breeding soundness of bulls. However, DIC was more effective in visualizing major defects, while BF (which included stained smear preparation) was considered to cause more minor defects. Thus DIC was considered to be the preferred method of semen assessment for accurate assessment of sperm morphology in bulls.
Assuntos
Bovinos , Microscopia/métodos , Espermatozoides/ultraestrutura , Acrossomo/ultraestrutura , Compostos de Anilina , Animais , Cruzamento , Corantes , Amarelo de Eosina-(YS) , Fertilidade , Masculino , Microscopia de Contraste de Fase , Estações do Ano , Contagem de Espermatozoides , Cabeça do Espermatozoide/ultraestrutura , Cauda do Espermatozoide/ultraestrutura , Espermatozoides/anormalidades , Espermatozoides/classificaçãoRESUMO
We analyzed the appearance and localization of the sub-acrosomal perinuclear theca (PT) during human spermiogenesis. The PT is tightly associated with acrosomal biogenesis.
Assuntos
Cabeça do Espermatozoide/fisiologia , Cabeça do Espermatozoide/ultraestrutura , Espermatogênese/fisiologia , Acrossomo/fisiologia , Acrossomo/ultraestrutura , Azoospermia/patologia , Núcleo Celular/fisiologia , Humanos , Masculino , Microscopia de Fluorescência , Morfogênese/fisiologia , Cabeça do Espermatozoide/patologiaRESUMO
OBJECTIVE: To evaluate fertilization and pregnancy rates, and pregnancy outcome in women diagnosed with non-determined infertility, and its correlation with sperm head morphology anomalies. MATERIAL AND METHODS: Retrospective study (ANOVA) including 126 patients diagnosed with non-determined infertility, attended from January 2002 to December 2006, admitted to embryo transference fertilization program, both conventional in vitro and intracytoplasmatic sperm injection. Spermatic morphology was analyzed in accordance with Kruger's strict criterion. All of patients had spermatic morphology higher than 5%, with 7.17% average. They were divided into three groups, in accordance with sperm head morphology anomalies percentage: group 1, less than 30%; group 2, 31 to 39%, and group 3, more than 40%. All of them had oocyte insemination: half conventional in vitro and half intracytoplasmatic sperm injection. RESULTS: Group 1 had higher percentage of fertilization, over groups 2 and 3 (65.44, 49.5 and 57.23%, respectively; p = 0.002). Abortion numbers showed meaningful differences too; group 1 had no abortions, meanwhile groups 2 and 3 had 30.77 and 22.22%, respectively. CONCLUSIONS: Evaluation of sperm head morphology anomalies must be a prognostic factor in pregnancy evolution, in women with nondetermined infertility.