RESUMO
Although the order Rodentia does not present a high risk of extinction compared to mammals as a whole, several families demonstrate high levels of threat and/or data deficiency, therefore highlighting the need for targeted research and the application of ecological and reproductive data to the development of conservation actions. The order Rodentia, the largest among mammals, includes 9 families, and the family Cricetidae is the most diverse of the Brazilian rodents. In Brazil, 12 of the 16 genera of Oecomys are found. Oecomys bicolor is known in Brazil as the 'arboreal rat' and is, found in dry, deciduous and tropical forests. The mean body weight of Oecomys bicolor was 35.8 g and the gonadal, tubular and epithelial somatic indexes were, 0.53%, 0.47% and 0.37%, respectively. Seminiferous tubules volume density was 89.72% and the mitotic and meiotic indexes corresponded to 8.59 and 2.45 cells, respectively, and the yield of spermatogenesis was 23.83 cells. The intertubular compartment represented 10.28% of the testis parenchyma and around 5% of the interstitial space was occupied by Leydig cells, whose number per gram of testis was 11.10 × 107 cells. By evaluating the biometric and histomorphometric characteristics of the testis, there is evidence that this species has a high investment in reproduction. Due to the high contribution of the seminiferous epithelium and the intertubular compartment in this species, compared to the others of the same family, it is possible to infer that the species Oecomys bicolor has a promiscuous reproductive behaviour.
Assuntos
Arvicolinae , Células Intersticiais do Testículo , Espermatogênese , Testículo , Animais , Espermatogênese/fisiologia , Masculino , Testículo/anatomia & histologia , Testículo/fisiologia , Células Intersticiais do Testículo/citologia , Células Intersticiais do Testículo/fisiologia , Arvicolinae/anatomia & histologia , Arvicolinae/fisiologia , Túbulos Seminíferos/anatomia & histologia , BrasilRESUMO
Adenine nucleotide translocator 4 (Ant4), an ATP/ADP transporter expressed in the early phases of spermatogenesis, plays a crucial role in male fertility. While Ant4 loss causes early arrest of meiosis and increased apoptosis of spermatogenic cells in male mice, its other potential functions in male fertility remain unexplored. Here, we utilized Ant4 knockout mice to delineate the effects of Ant4-deficiency on male reproduction. Our observations demonstrated that Ant4-deficiency led to infertility and impaired testicular development, which was further investigated by evaluating testicular oxidative stress, autophagy, and inflammation. Specifically, the loss of Ant4 led to an imbalance of oxidation and antioxidants. Significant ultrastructural alterations were identified in the testicular tissues of Ant4-deficient mice, including swelling of mitochondria, loss of cristae, and accumulation of autophagosomes. Our results also showed that autophagic flux and AKT-AMPK-mTOR signaling pathway were affected in Ant4-deficient mice. Moreover, Ant4 loss increased the expression of pro-inflammatory factors. Overall, our findings underscored the importance of Ant4 in regulating oxidative stress, autophagy, and inflammation in testicular tissues. Taken together, these insights provided a nuanced understanding of the significance of Ant4 in testicular development.
Assuntos
Infertilidade Masculina , Translocases Mitocondriais de ADP e ATP , Estresse Oxidativo , Testículo , Animais , Masculino , Camundongos , Apoptose/fisiologia , Autofagia/fisiologia , Infertilidade Masculina/metabolismo , Camundongos Knockout , Translocases Mitocondriais de ADP e ATP/metabolismo , Translocases Mitocondriais de ADP e ATP/genética , Estresse Oxidativo/fisiologia , Transdução de Sinais/fisiologia , Espermatogênese/fisiologia , Testículo/metabolismo , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismoRESUMO
Context A maternal high-fat diet is thought to pose a risk to spermatogenesis in the progeny. Aims We tested whether a maternal high-fat diet would affect Sertoli cell expression of transcription factors (insulin-like growth factor I (IGF-I); glial-cell line-derived neurotrophic factor (GDNF); Ets variant 5 (ETV5)) and cell proliferation and apoptotic proteins, in the testis of adult offspring. Methods Pregnant rats were fed ad libitum with a standard diet (Control) or a high-fat diet (HFat) throughout pregnancy and lactation. After weaning, male pups were fed the standard diet until postnatal day 160. Males were monitored daily from postnatal day 34 to determine onset of puberty. On postnatal day 160, their testes were processed for morphometry and immunohistochemistry. Key results The HFat diet increased seminiferous-tubule diameter (P P P P P P P P Conclusions A maternal high-fat diet alters the balance between spermatogonia proliferation and spermatid apoptosis. Implications A maternal high-fat diet seems to 'program' adult male fertility.
Assuntos
Apoptose , Proliferação de Células , Dieta Hiperlipídica , Lactação , Fenômenos Fisiológicos da Nutrição Materna , Efeitos Tardios da Exposição Pré-Natal , Testículo , Animais , Feminino , Masculino , Gravidez , Apoptose/fisiologia , Lactação/fisiologia , Testículo/metabolismo , Testículo/patologia , Ratos , Efeitos Tardios da Exposição Pré-Natal/patologia , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Fenômenos Fisiológicos da Nutrição Materna/fisiologia , Espermatogênese/fisiologia , Células de Sertoli/metabolismo , Células de Sertoli/patologia , Fator de Crescimento Insulin-Like I/metabolismo , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Ratos WistarRESUMO
BACKGROUND: Spermatogonial stem cells (SSCs) are critical for sustaining spermatogenesis. Even though several regulators of SSC have been identified in rodents, the regulatory mechanism of SSC in humans has yet to be discovered. METHODS: To explore the regulatory mechanisms of human SSCs, we analyzed publicly available human testicular single-cell sequencing data and found that Ankyrin repeat and SOCS box protein 9 (ASB9) is highly expressed in SSCs. We examined the expression localization of ASB9 using immunohistochemistry and overexpressed ASB9 in human SSC lines to explore its role in SSC proliferation and apoptosis. Meanwhile, we used immunoprecipitation to find the target protein of ASB9 and verified its functions. In addition, we examined the changes in the distribution of ASB9 in non-obstructive azoospermia (NOA) patients using Western blot and immunofluorescence. RESULTS: The results of uniform manifold approximation and projection (UMAP) clustering and pseudotime analysis showed that ASB9 was highly expressed in SSCs, and its expression gradually increased during development. The immunohistochemical and dual-color immunofluorescence results displayed that ASB9 was mainly expressed in nonproliferating SSCs. Overexpression of ASB9 in the SSC line revealed significant inhibition of cell proliferation and increased apoptosis. We predicted the target proteins of ASB9 and verified that hypoxia-inducible factor 1-alpha inhibitor (HIF1AN), but not creatine kinase B-type (CKB), has a direct interaction with ASB9 in human SSC line using protein immunoprecipitation experiments. Subsequently, we re-expressed HIF1AN in ASB9 overexpressing cells and found that HIF1AN reversed the proliferative and apoptotic changes induced by ASB9 overexpression. In addition, we found that ABS9 was significantly downregulated in some NOA patients, implying a correlation between ASB9 dysregulation and impaired spermatogenesis. CONCLUSION: ASB9 is predominantly expressed in human SSCs, it affects the proliferation and apoptotic process of the SSC line through HIF1AN, and its abnormal expression may be associated with NOA.
Assuntos
Testículo , Ubiquitina-Proteína Ligases , Masculino , Humanos , Ubiquitina-Proteína Ligases/metabolismo , Testículo/metabolismo , Espermatogênese/fisiologia , Linhagem Celular , Proliferação de Células , Apoptose , Ubiquitinas/metabolismo , Oxigenases de Função Mista/metabolismo , Proteínas Repressoras/metabolismo , Proteínas Supressoras da Sinalização de Citocina/metabolismoRESUMO
BACKGROUND: Spermatogonial stem cells (SSCs) are critical for sustaining spermatogenesis. Even though several regulators of SSC have been identified in rodents, the regulatory mechanism of SSC in humans has yet to be discovered. METHODS: To explore the regulatory mechanisms of human SSCs, we analyzed publicly available human testicular single-cell sequencing data and found that Ankyrin repeat and SOCS box protein 9 (ASB9) is highly expressed in SSCs. We examined the expression localization of ASB9 using immunohistochemistry and overexpressed ASB9 in human SSC lines to explore its role in SSC proliferation and apoptosis. Meanwhile, we used immunoprecipitation to find the target protein of ASB9 and verified its functions. In addition, we examined the changes in the distribution of ASB9 in non-obstructive azoospermia (NOA) patients using Western blot and immunofluorescence. RESULTS: The results of uniform manifold approximation and projection (UMAP) clustering and pseudotime analysis showed that ASB9 was highly expressed in SSCs, and its expression gradually increased during development. The immunohistochemical and dual-color immunofluorescence results displayed that ASB9 was mainly expressed in nonproliferating SSCs. Overexpression of ASB9 in the SSC line revealed significant inhibition of cell proliferation and increased apoptosis. We predicted the target proteins of ASB9 and verified that hypoxia-inducible factor 1-alpha inhibitor (HIF1AN), but not creatine kinase B-type (CKB), has a direct interaction with ASB9 in human SSC line using protein immunoprecipitation experiments. Subsequently, we re-expressed HIF1AN in ASB9 overexpressing cells and found that HIF1AN reversed the proliferative and apoptotic changes induced by ASB9 overexpression. In addition, we found that ABS9 was significantly downregulated in some NOA patients, implying a correlation between ASB9 dysregulation and impaired spermatogenesis. CONCLUSION: ASB9 is predominantly expressed in human SSCs, it affects the proliferation and apoptotic process of the SSC line through HIF1AN, and its abnormal expression may be associated with NOA.
Assuntos
Humanos , Masculino , Testículo/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Proteínas Repressoras/metabolismo , Espermatogênese/fisiologia , Ubiquitinas/metabolismo , Linhagem Celular , Apoptose , Proliferação de Células , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Oxigenases de Função Mista/metabolismoRESUMO
A espermiogênese é o processo final da espermatogênese no qual a espermátide se transforma em espermatozoide. Durante esse processo podem ocorrer alterações espermáticas, especialmente no acrossoma, na morfologia da cabeça, na condensação da cromatina e na formação dos vacúolos nucleares. A diferenciação entre os quadros clínicos é feita com repetições dos exames andrológicos. A avaliação morfológica indica a situação da espermiogênese nas 4 semanas anteriores a coleta do sêmen. Com os resultados do exame andrológico é possível identificar a qualidade seminal naquele período. O presente artigo é o relato de caso de um touro jovem doador de sêmen com 26 meses de idade, no início do regime de coletas de sêmen em um Centro de Coleta e Processamento de Sêmen no Sul do Brasil. A produção espermática deste animal foi avaliada durante 12 meses de coleta, apresentando sempre morfologia espermática muito alterada e motilidade média de 54% no exame imediato. Em 13 coletas seminais, o ejaculado deste animal apresentou em média 86% de defeitos maiores, 10% de defeitos menores e 96% de defeitos totais. Os defeitos maiores, que são os que possuem maior efeito na fertilidade, estão diretamente ligados a espermiogênese e os defeitos menores, que possuem menor efeito na fertilidade, são ocasionados principalmente durante o trânsito pelo epidídimo. O quadro clínico desse animal demonstrou a importância do exame morfológico para avaliar a espermatogênese e principalmente o processo de diferenciação final da espermátide em espermatozoide. Casos como este devem ser descritos como espermiogênese alterada e o reprodutor deve ser afastado da reprodução, após descartar alterações morfológicas devido a degeneração testicular grave.(AU)
Spermiogenesis is the final process of spermatogenesis in which the spermatid transforms into sperm. During this process, sperm alterations may occur, especially in the acrosome, head morphology, chromatin condensation and formation of nuclear vacuoles. Differentiation between clinical conditions is made with repetitions of andrological exams. The morphological evaluation indicates the status of spermiogenesis in the 4 weeks prior to semen collection. With the results of the breeding soundness examination, it is possible to identify the seminal quality in that period. The present article is a case report of a young 26-month-old semen donor bull, that started semen collection routines at a Semen Collection and Processing Center in southern Brazil. The sperm production of this animal was evaluated during 12 months of collection, always showing very altered sperm morphology and average motility of 54% in the immediate examination. In 13 seminal collections, the ejaculate of this animal presented an average of 86% of major defects, 10% of minor defects and 96% of total defects. Major defects, which have the greatest impact on fertility, are directly linked to spermiogenesis and minor defects, which have less effect on fertility, are mainly caused during transit through the epididymis. The clinical conditions of this animal demonstrated the importance of the morphological examination to evaluate spermatogenesis and especially the process of final differentiation of the spermatid into spermatozoa. Cases like this should be described as altered spermiogenesis and the bull should be withdrawn from breeding, after ruling out morphological changes due to severe testicular degeneration.(AU)
Assuntos
Animais , Masculino , Espermatogênese/fisiologia , Bovinos/embriologia , Análise do Sêmen/veterináriaRESUMO
Given the importance of males as semen donors for artificial insemination (AI) and the high incidence of low birthweight piglets at commercial farms, the impact of birthweight on fertility in boars deserves special attention. The aim of this study was to evaluate testicular morphofunctional parameters and semen characteristics in different birthweight boars. Forty littermate males were selected at birth and divided into two experimental groups, according to birthweight: high (HW, birthweight ranging from 1.80 to 2.15 kg, n = 20) and low (LW, birthweight ranging from 0.75 to 1.10 kg, n = 20). At 170 days of age, a sub-set of 24 littermate boars (n = 12 HW and n = 12 LW) were randomly selected for semen collection, which was performed once a week, at a 15-day interval, during five weeks. At 300 days of age, boars were orchiectomized, and the testis processed for histological and molecular analyses. The HW group was heavier and presented larger testes compared to LW animals (P < 0.05). Despite that, birthweight did not significantly affect semen volume or sperm quality parameters (concentration, motility, vigor or morphology), although LW boars produced 38.2% fewer total sperm and 24% lower semen concentration, leading to 36.8% less inseminating doses. The histomorphometrical evaluation showed that seminiferous tubules diameter and germinal epithelium height were similar between experimental groups. However, LW boars presented shorter seminiferous tubules and, consequently, fewer Sertoli cells per testis (P 0.05). Even though plasma testosterone levels were equivalent in both birthweight groups, LW testis presented less androgen receptors (P < 0.05). Additionally, birthweight was positively correlated with total seminiferous tubule length and number of Sertoli cells (P < 0.01), and with body and testis weights (P < 0.01). Taken together, even though adult LW boars showed no evident semen pathologies or spermatogenesis commitment, mature HW males have the potential to produce more spermatozoa, consequently more semen doses per ejaculate, being more valuable to an industry that relies on AI.
Assuntos
Peso ao Nascer , Sêmen , Testículo , Animais , Peso ao Nascer/fisiologia , Masculino , Sêmen/fisiologia , Contagem de Espermatozoides/veterinária , Espermatogênese/fisiologia , Suínos , Testículo/anatomia & histologia , Testículo/fisiologiaRESUMO
In mammals, spermatogenesis is a complex and cyclic process in which a spermatogonia turns into a highly differentiated cell: the spermatozoa. Spermatogenesis comprises proliferation of spermatogonia (spermatocytogenesis), meiosis of spermatocytes and finally differentiation of spermatids into spermatozoa (spermiogenesis). This review summarizes the current knowledge on domestic cat spermatogenesis including its physiology, development, efficiency and pathologies as well as their novel non-invasive diagnostic methods. This information will provide a resource for further studies to achieve precise fundamental knowledge of key aspects that will facilitate breeding, management and contraception in this popular species.
Assuntos
Espermátides , Espermatogênese , Animais , Gatos , Masculino , Mamíferos , Meiose , Espermátides/fisiologia , Espermatócitos/fisiologia , Espermatogênese/fisiologia , Espermatogônias , Espermatozoides/fisiologia , TestículoRESUMO
Os hermafroditas verdadeiros são animais com intersexualidade, nos quais as gônadas masculinas e femininas se desenvolvem simultaneamente no mesmo indivíduo. Este trabalho relata um caso de hermafrodita na espécie suína após descarte de uma fêmea por diagnóstico de anestro. Tratava-se de uma fêmea com 222 dias de idade e 150 kg a qual havia sido selecionada para reprodução e permanecia em anestro. O indivíduo foi descartado e enviado para o abate em frigorífico sob inspeção federal. Após o abate foi observado no sistema genital aspectos anatômicos sugestivos de hermafroditismo. O sistema genital foi coletado, inspecionado macroscopicamente e conservado em formol a 10% para avaliação microscópica. Durante a análise macroscópica foi verificada a presença de vulva e vagina, contendo um pênis rudimentar. O útero, a cérvix, os cornos uterinos e oviduto estavam presentes e com ausência de anormalidade. Havia um ovário esquerdo com aspectos morfológicos de ciclicidade e folículos contendo oócitos de alta qualidade. À direita, entretanto, foi constatada a presença de um testículo, epidídimo e plexo pampiniforme. A avaliação histopatológica comprovou os achados macroscópicos, demonstrando um ovário contendo folículos, corpos lúteos e cistos foliculares; plexo pampiniforme; testículo e epidídimo com aspecto histológico normal, porém, com ausência de células espermatogênicas. Assim, com base nos achados macro e microscópicos descritos nesse relato de caso, o indivíduo foi considerado como hermafrodita verdadeiro.(AU)
True hermaphrodites are intersexual animals, in which both male and female gonads develop simultaneously in the same individual. This paper reports a case of a true hermaphrodite in the swine species after its culling for anestrus. The specimen was a female of 222 days of age and 150 kg of body weight, that was selected for reproduction and was permanently in anestrus. This individual was culled and sent to an abattoir under federal inspection. After slaughtering, it was observed in the reproductive tract anatomical aspects that suggested hermaphroditism. The reproductive tract was collected, evaluated macroscopically, and preserved in formaldehyde at 10 % to be evaluated by light microscopy. During the macroscopic analysis, the presence of a vulva and a vagina with a rudimentary penis was detected. The uterus, the cervix, uterine horns, and the oviduct were present and abnormalities were not detected. The left ovary exhibited morphological signs of cyclicity and contained follicles with high quality oocytes. On the right side, however, it was observed a testicle, epididymis, and a pampiniform plexus. The histopathological assessment corroborated the macroscopic findings, which demonstrated that the ovary had follicles, corpus luteum, and follicular cysts, and pampiniform plexus, testicle and epididymis were histologically normal, though with the absence of spermatogenic cells. Thus, based on the macro and microscopic findings described herein, the individual was considered a true hermaphrodite.(AU)
Assuntos
Animais , Feminino , Transtornos do Desenvolvimento Sexual/diagnóstico , Suínos/anatomia & histologia , Anestro/fisiologia , Espermatogênese/fisiologia , Genitália/fisiologiaRESUMO
A morfologia espermática é uma característica seminal que está associada à fertilidade e é, portanto, um componente importante na rotina do controle de qualidade em Centros de Coleta e Processamento de Sêmen (CCPS). É utilizada para caracterizar e quantificar os vários tipos de anormalidades nas células espermáticas do touro, usando preparação em montagem úmida para microscopia de Contraste de Interferência Diferencial (DIC), Contraste de Fase (CF) e Lâmina Corada (LC). O perfil da morfologia espermática permite a tomada de decisão sobre o aproveitamento ou descarte de um ejaculado para criopreservação e predizer a fertilidade de uma amostra de sêmen. Alterações no espermograma, com evidências do histórico e exame físico do touro, levam a interpretação da função testicular anormal, e consequentemente, ao diagnóstico, tratamento e prognóstico, se for o caso. As causas mais comuns de uma espermatogênese anormal nos touros incluem: termorregulação testicular anormal, desequilíbrios hormonais, estresse de manejo, ambiental e nutricional, efeitos de toxinas ou expressão de genes deletérios, além de defeitos de origem genética. Conhecendo-se a estrutura do espermatozoide, torna-se mais fácil a identificação das anormalidades espermáticas bem como a descrição das mesmas e uma melhor compreensão de sua origem.(AU)
Sperm morphology is a seminal trait that is associated with fertility and is therefore an important component of routine quality control in Semen Collection and Processing Centers (SCPC). It is used to characterize and quantify the various types of abnormalities in bull sperm cells, using wet mount preparation for Differential Interference Contrast (DIC), Phase Contrast (PC) and Slides Smear (SS) microscopy. The sperm morphology profile allows decision making on the use or disposal of an ejaculate for cryopreservation and predicts the fertility of a semen sample. Changes in sperm morphology may indicate that fertility may be impacted and therefore it is important to routinely monitor ejaculates. An abnormal spermogram with evidence from a history and physical examination of the bull can lead to the interpretation of abnormal testicular function, and consequently, to the diagnosis, treatment and prognosis, if any. The most common causes of abnormal spermatogenesis in bulls include: abnormal testicular thermoregulation, hormonal imbalances, management, environmental and nutritional stress, effects of toxins or expression of deleterious genes, in addition to defects of genetic origin. Knowledge in the structure of the sperm makes easier the identification of sperm abnormalities, as well as their description and provides a better understanding of their origin.(AU)