RESUMO
The molecular mechanisms regulating follicular development and ensuring primordial follicle activation remain undefined. To help elucidate these mechanisms, this proteomic study of bovine ovarian tissue identified the differential molecular profiles of preantral follicles together with the spatial distribution of the most abundant molecular components in the tissue. Isolated primordial, primary and secondary follicles were individually placed on a MALDI target plate for mass spectral acquisitions, with detection of different m/z ranges. Ovarian tissue was sectioned and analysed in the m/z 400-2,000 range. Results of the first analysis indicated a similarity pattern in the molecular protein profile among different follicular classes in the m/z ranges of 100-1000 and 25,000-200,000, but in the m/z ranges of 800-4000, 4000-20,000 and 15,000-70,000, primary and secondary follicles shared similar clustering profiles which were different from primordial follicles (p < .05). In the second analysis, it was possible to correlate some intense molecular components in the tissue from global mass spectrum with the ions detected in the first analysis. Molecular components at m/z 11,325 (±230) were also detected in primary and secondary follicles in the experiment with isolated follicles, in addition to ions at m/z 4,029 (±120), 13,799 (±70), 5,547 (±9), 15,313 (±200), 7,018 (±40) and 7,663 (±90) which were also intensely detected in primary and secondary follicles. The present proteomic approaches evaluated different mass ranges of preantral follicles in bovine ovarian tissue and also indicated the spatial distribution of the most abundant molecular components. This study hopes to pave the way for future research identifying and characterizing specific proteins involved in follicle activation in bovine follicles, in order to better understand folliculogenesis and potentially improve mammalian follicle culture systems.
Assuntos
Folículo Ovariano , Proteômica , Animais , Bovinos , Feminino , Ovário , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterináriaRESUMO
Controlling populations of free-roaming dogs and cats poses a huge challenge worldwide. Non-surgical neutering strategies for male animals have been long pursued, but the implementation of the procedures developed has remained limited to date. As submitting the testes to high temperatures impairs spermatogenesis, the present study investigated localized application of magnetic nanoparticle hyperthermia (MNH) to the testicles as a potential non-surgical sterilization method for animals. An intratesticular injection of a magnetic fluid composed of manganese-ferrite nanoparticles functionalized with citrate was administered followed by testicle exposure to an alternate magnetic field to generate localized heat. Testicular MNH was highly effective, causing progressive seminiferous tubule degeneration followed by substitution of the parenchyma with stromal tissue and gonadal atrophy, suggesting an irreversible process with few side effects to general animal health.
RESUMO
This study aimed to evaluate the effects of an intratesticular injection of silver nanoparticles (AgNPs) on reproductive parameters and health of rats, and to evaluate the AgNPs biodistribution in order to develop a nanotechnological contraceptive agent for male animals. Treated animals received 220 µL of AgNPs solution (0.46 µg-Ag/ml) in each testicle and were euthanized: seven, 14, 28, and 56 days after injection. A significant decrease (p < 0.05) in the percentage of motile sperm in D7 (8.8%) was observed, comparing to the control (73.3%), D14 (86.0%), D28 (68.2%), and D56 (90.0%) groups. D7 group also presented a decrease (p < 0.05) in the percentage of normal spermatozoa. Additionally, D7 group showed an increase (p < 0.05) in abnormal midpiece and sperm head morphology compared to the Control group. Seminiferous tubules presented all germline cell types and spermatozoa for all groups. However, D7 group did not present spermatozoa in the epididymis, whereas some spermatozoa and cellular debris were visible in D14 and D28 groups. All animals presented hematological parameters, creatinine, and alanine aminotransferase values within the normal limits for Wistar rats. The percentage of silver found in the liver was always higher than in the other organs analyzed. A pioneering mathematical model is proposed, from which the half-life time of silver in the liver (17 days), spleen (23 days), lungs (30 days), and kidneys (35 days) was extracted. In conclusion, some acute and severe toxic effects were observed in sperm cells following intratesticular injection of AgNPs, although these effects were reversible. No adverse effects to general animal health were observed.
Assuntos
Nanopartículas Metálicas/toxicidade , Reprodução/efeitos dos fármacos , Prata/toxicidade , Espermatozoides/efeitos dos fármacos , Testículo/efeitos dos fármacos , Alanina Transaminase/metabolismo , Animais , Epididimo/efeitos dos fármacos , Epididimo/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Nanopartículas Metálicas/administração & dosagem , Ratos , Ratos Wistar , Prata/administração & dosagem , Prata/farmacocinética , Espermatozoides/metabolismo , Baço/efeitos dos fármacos , Baço/metabolismo , Testículo/metabolismo , Distribuição TecidualRESUMO
The present study evaluated revascularization time of fresh and cryopreserved cat ovarian tissue after transplantation to subcutaneous tissue. Ovaries of five cats were used and eight pieces of ovarian tissue were taken from each pair of ovaries. Immediately after removal, three pieces were transplanted and one fixed for fresh control. The remaining four pieces were cryopreserved and, after thawing, one was fixed for cryopreservation control and three were transplanted. Grafts were recovered on days 2 (D2), 4 (D4) and 6 (D6) post-transplantation. Blood vessels were identified by immunohistochemistry and doppler ultrasound. Immunohistochemistry showed that the percentages of total tissue area occupied by blood vessels were similar (P > 0.05) in fresh and cryopreserved tissues. In both cases, blood vessel area was significantly higher (P < 0.05) on D4 and D6 compared to D0. Ultrasound analysis showed vascularization improvement on the periphery of grafts from D2 to D4 and from D4 to D6, both in fresh and cryopreserved tissue samples. Nonetheless, there was a significant decrease (P < 0.05) in the percentage of morphologically normal follicles (MNF) after transplantation compared to non-transplanted tissue (D0), both for fresh and cryopreserved samples. Moreover, the number of follicles found in samples was considerably smaller after grafting. In conclusion, revascularization of ovarian tissue autotransplanted to subcutaneous tissue in domestic cats occurs within 4 days after transplantation, both for fresh and cryopreserved tissue. However, large follicular loss has been observed in the first days post-transplantation, especially in cryopreserved tissues.
Assuntos
Criopreservação/veterinária , Ovário/irrigação sanguínea , Ovário/transplante , Animais , Gatos , Feminino , Folículo Ovariano/transplante , Fatores de Tempo , Ultrassonografia DopplerRESUMO
The aim of this study was to analyse the reproductive aspects of male bats of three common species of the Phyllostomidae family: Artibeus lituratus, Platyrrhinus lineatus and Sturnira lilium, during dry and rainy months in a specific area of the Cerrado biome. Body weight was significantly higher during the dry months for S. lilium. The gonadosomatic index (GSI) and testicular weight were not significantly different between dry and rainy periods. The tubular parameters were significantly bigger in A. lituratus than in the other two species during both periods. No difference in the tubular/interstitial ratio was observed in any of the species during both periods. In both periods, all sperm cells and germ cell developmental stages were visible on seminiferous tubules whereas sperm cells were observed in epididymides of all sampled animals. The percentage of morphologically normal sperm was low (35%-60%), with no difference between periods. Spermatozoa from A. lituratus presented a leaf-shaped head, while the head was round-shaped in the other two species. In conclusion, our data suggest that males from the three studied species did not present reproductive latency during the most critical weather periods (dry and rainy months) in the metropolitan region of Brasilia, Brazil.
Assuntos
Quirópteros/anatomia & histologia , Espermatozoides , Testículo , Animais , Brasil , Masculino , Chuva , Reprodução , Estações do Ano , Túbulos Seminíferos/anatomia & histologia , Espermatogênese , Espermatozoides/anormalidades , Espermatozoides/citologia , Testículo/anatomia & histologia , Testículo/citologiaRESUMO
Magnetic nanoparticles can be used in different areas of biology. It is therefore important to know the effects of such nanomaterials on germline cells as they may traverse the blood-testis barrier. This work aimed to evaluate the response of bull sperm after exposure to a magnetic fluid containing DMSA-coated maghemite nanoparticles (MNP-DMSA) in order to determine nanotoxicity. Bull sperm was incubated with MNP-DMSA at final concentrations of 0.06, 0.03 or 0.015 mg Fe/mL. Sperm kinetics, plasma membrane integrity and acrosome reaction were evaluated over a 4 h incubation period. The sperm cells were also evaluated by transmission electron microscopy. Exposure of bull sperm to MNP-DMSA did not affect sperm kinetics or integrity. Neither ultrastructural damage of sperm cells nor uptake of nanoparticles by the spermatozoa was observed. In conclusion, MNP-DMSA does not affect sperm function or structure under the conditions tested.
Assuntos
Compostos Férricos/administração & dosagem , Nanopartículas Metálicas/administração & dosagem , Espermatozoides/efeitos dos fármacos , Succímero/administração & dosagem , Animais , Bovinos , Membrana Celular/efeitos dos fármacos , Compostos Férricos/química , Masculino , Nanopartículas Metálicas/química , Microscopia Eletrônica de Transmissão , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Espermatozoides/ultraestrutura , Succímero/químicaRESUMO
Melanoma is the most aggressive and lethal form of skin cancer, responsible for >80% of deaths. Standard treatments for late-stage melanoma usually present poor results, leading to life-threatening side effects and low overall survival. Thus, it is necessary to rethink treatment strategies and design new tools for the treatment of this disease. On that ground, we hereby report the use of acai oil in nanoemulsion (NanoA) as a novel photosensitizer for photodynamic therapy (PDT) used to treat melanoma in in vitro and in vivo experimental models. NIH/3T3 normal cells and B16F10 melanoma cell lines were treated with PDT and presented 85% cell death for melanoma cells, while maintaining high viability in normal cells. Flow cytometry indicated that cell death occurred by late apoptosis/necrosis. Tumor bearing C57BL/6 mice treated five times with PDT using acai oil in nanoemulsion showed tumor volume reduction of 82% in comparison to control/tumor group. Necrotic tissue per tumor area reached its highest value in PDT-treated mice, supporting PDT efficacy. Overall, acai oil in nanoemulsion was an effective photosensitizer, representing a promising source of new photosensitizing molecules for PDT treatment of melanoma, a tumor with an inherent tendency to be refractory for this type of therapy.
Assuntos
Emulsões , Euterpe/química , Melanoma/tratamento farmacológico , Fotoquimioterapia , Óleos de Plantas/uso terapêutico , Animais , Camundongos , Camundongos Endogâmicos C57BL , Células NIH 3T3 , NanotecnologiaRESUMO
In this study, a multivariate analysis of morphological and physiological characteristics was performed on clinically healthy rams from six breeds (Santa Ines, Bergamasca, Dorper, Texel, Ile de France and Hampshire Down) to determine if these characteristics were able to separate and determine the most important variables in the differentiation of breeds for heat adaptation. To characterize the thermal environment, mean temperature was 23°C and relative humidity ranged between 30.6-55.6%. Morphological and physiological data were subjected to multivariate statistical tests including principal components (PRINCOMP), clustering (CLUSTER), discriminant (DISCRIM), step-bystep (STEPDISC) and canonical (CANDISC) analyses, using the Statistical Analysis System (SAS®). A multiple analysis of variance (MANOVA) was carried out with the variables defined as important by the discriminant analysis. The principal components analysis for biometric characteristics and scrotum-testicle, for physiological characteristics and body temperature as well as the characteristics of the skin and hair explained 60, 70 and 67 % of the total variation, respectively. The dendrogram showed a clear separation between the breeds studied and the existence of two distinct groups, one formed by the Texel and the other by the other breeds, considering all the characteristics used in the study. The most useful morphological parameters to explain heat tolerance were diameter of hair, layer thickness of hair at withers, 12th thoracic vertebra and rump, withers height, thoracic and scrotal circumferences, body weight, anterior and posterior shin perimeters, hair and epidermis brightness as well as the content of red and yellow in the epidermis. Among physiological characteristics, respiratory rate was better than rectal temperature and heart rate to explain changes caused by thermal stress. From the multivariate and variance analyzes it can be concluded that the Santa Ines breed was the most tolerant to heat stress as it presented a highly pigmented epidermis, a shorter hair of larger diameter, the lower layer thickness of hair at withers, 12th thoracic vertebra and rump, the lower temperatures in the testicle and at the 12th thoracic vertebra as well as the lower respiratory rate and rectal temperature value.
Neste estudo, foram realizadas análises multivariadas das características fisiológicas e morfológicas em carneiros clinicamente saudáveis de seis raças (Santa Inês, Bergamácia, Dorper, Texel, Ile de France e Hampshire Down) para determinar se essas características foram capazes de separar as raças e determinar as variáveis mais importantes na diferenciação das raças na adaptação ao calor. Os dados foram submetidos a testes estatísticos multivariados, incluindo as análises de componentes principais (PINCOMP), agrupamento (CLUSTER), discriminante (DISCRIM), step-by-step (STEPDISC) e canônica (CANDISC), utilizando o pacote estatístico SAS®. A análise variância de múltipla (MANOVA) foi realizada com as variáveis definidas como importante pela análise discriminante. A análise dos componentes principais para características biométricas e escroto-testiculares, para as características fisiológicas e de temperatura corporal e para as características da pele e pelo explicaram 60, 70 e 67% da variação total, respectivamente. O dendrograma mostrou uma clara separação entre as raças estudadas e a existência de dois grupos distintos, um formado pela raça Texel e o outro pelas raças Dorper, Hampshire Down, Ile de France, Santa Inês e Bergamácia, considerando todas as características avaliadas. As características morfológicas mais importantes para explicar a tolerância ao calor foram o diâmetro do pelo, as espessuras das camadas de pelo na cernelha, na décima segunda vértebra torácica e na garupa, a altura da cernelha, as circunferências torácica e escrotal, o peso corporal, os perímetros das canelas anterior e posterior, as luminosidades da pele e do pelo, bem como os teores de pigmentação vermelho e amarelo na epiderme. Entre as características fisiológicas a frequência respiratória foi melhor que a temperatura retal e a frequência cardíaca para explicar as mudanças causadas pelo estresse térmico. A partir das análises multivariada e de variância pode-se concluir que a raça Santa Inês foi a mais tolerante ao estresse térmico, uma vez que apresentou epiderme altamente pigmentada, pelo mais curto e de diâmetro maior, menores espessuras das camadas de pelo na cernelha, na décima segunda vértebra torácica e na garupa, menores temperaturas no testículo e na décima segunda vértebra torácica, bem como as menores frequência respiratória e temperatura retal.
Assuntos
Ovinos , Termotolerância , Temperatura AltaRESUMO
Cottonseed coproducts contain gossypol which can have specific effects upon the male reproductive system. We evaluated the reproductive development of male lambs close to puberty fed on three cotton coproducts. Twenty-four 5-month old male lambs received four diets: 20 % of dry matter intake of whole cottonseed (WCS), high oil cottonseed meal (CSC), cottonseed meal (CSM), and a control group (CTL) without any cottonseed coproducts. Free gossypol intake was 16.32, 6.98, 5.47 and 0 mg kg1 BW1 d1 for WCS, CSC, CSM and CTL, respectively. Every 15 days, the animals were weighted and serum and semen samples were collected. After 95 days, testis samples were collected for analysis under light and transmission electron microscopes. The CTL group had higher testosterone concentrations than CSC at the end of the trial and lower total sperm defects, higher mass movement and higher scores for seminiferous epithelium than other treatments. The WCS and CSC groups showed higher levels of segmental aplasia lesion in sperm than other diets, which showed that high levels of gossypol led to higher occurrence of this lesion. Cottonseed coproducts had a negative impact on the reproductive system of pubertal lambs regardless of gossypol concentration. Therefore, the use of cottonseed coproducts to feed lambs earmarked for reproduction is not safe.
Assuntos
Masculino , Animais , Fenômenos Reprodutivos Fisiológicos , Gossipol , Gossypium , Ovinos , Sementes , Soro , Sêmen , Testosterona , TestículoRESUMO
Cottonseed coproducts contain gossypol which can have specific effects upon the male reproductive system. We evaluated the reproductive development of male lambs close to puberty fed on three cotton coproducts. Twenty-four 5-month old male lambs received four diets: 20 % of dry matter intake of whole cottonseed (WCS), high oil cottonseed meal (CSC), cottonseed meal (CSM), and a control group (CTL) without any cottonseed coproducts. Free gossypol intake was 16.32, 6.98, 5.47 and 0 mg kg1 BW1 d1 for WCS, CSC, CSM and CTL, respectively. Every 15 days, the animals were weighted and serum and semen samples were collected. After 95 days, testis samples were collected for analysis under light and transmission electron microscopes. The CTL group had higher testosterone concentrations than CSC at the end of the trial and lower total sperm defects, higher mass movement and higher scores for seminiferous epithelium than other treatments. The WCS and CSC groups showed higher levels of segmental aplasia lesion in sperm than other diets, which showed that high levels of gossypol led to higher occurrence of this lesion. Cottonseed coproducts had a negative impact on the reproductive system of pubertal lambs regardless of gossypol concentration. Therefore, the use of cottonseed coproducts to feed lambs earmarked for reproduction is not safe.(AU)