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Abstract Introduction: Testicular histology constitutes one of the least explored aspects in frogs of the genus Atelopus. This taxonomic group shows an alarming population decline; therefore, its reproductive biology is one of the greatest topics of interest for its conservation. Objective: To describe the testicular morphology and the spermatogenetic lineage cells in adult males of Atelopus laetissimus, Atelopus nahumae, and Atelopus carrikeri in the Sierra Nevada de Santa Marta, Colombia. Methods: During June - July 2017 and 2018, sampling was conducted in the localities of San Lorenzo and Páramo Cebolletas, Sierra Nevada de Santa Marta (SNSM), to collect 15 adult males, 5 per species. Testes samples were fixed in Bouin to be processed by the standard paraffin-embedding technique. Histological sections (3 μm) were stained with Hematoxylin-eosin and Mallory-Heidenhain-Azan-Gomori's. For the description and photographic register of the germ cells, the photonic microscopy technique was used with the differential interference contrast system. Results: The testes are oval organs, compact, light yellow color, and with little vascularization. Externally, they are surrounded by a thin albuginea tunic constituted by regular dense connective tissue. Inside this layer, they are composed of numerous seminiferous tubules of hexagonal contour, in which germ cell cysts are distinguished at different stages of spermatogenesis (spermatogonia I and II, spermatocyte I and II, and early and late spermatids) and spermiogenesis (spermatozoa in fascicles and free spermatozoa). Separating the seminiferous structures is the interstitial tissue in which Leydig cells and blood vessels stand out. Additionally, in the cranial part of the testis, the Bidder's organ was found, formed by two distinguishable regions, the cortex and the medulla. In the cortex, there are previtellogénic oocytes of different sizes surrounded by a monolayer of flat follicular cells. For its part, the medullary region is the connective tissue that nourishes the oocytes and is constituted by blood capillaries. Conclusions: The gonads of the three species analyzed present a cystic cellular organization similar to other anurans, where all stages of spermatogenesis and spermiogenesis were identified, possibly indicating a continuous reproductive activity. Likewise, the Bidder's organ is reported for the first time in the three Atelopus species, which allows suggesting a possible sexual reversion in case of a population decrease of females as a reproductive strategy.
Resumen Introducción: La histología testicular constituye uno de los aspectos menos explorados en las ranas del género Atelopus. Este grupo taxonómico ostenta un declive poblacional alarmarte, es por ello, que su biología reproductiva resulta uno de los temas de mayor interés para su conservación. Objetivo: Describir la morfología testicular y las células del linaje espermatogénico en machos adultos de Atelopus laetissimus, Atelopus nahumae y Atelopus carrikeri en la Sierra Nevada de Santa Marta, Colombia. Métodos: Durante Junio - Julio de 2017 y 2018 se realizaron muestreos en las localidades de San Lorenzo y Páramo Cebolletas, Sierra Nevada de Santa Marta (SNSM), para recolectar 15 machos adultos, 5 por especie. Las muestras de testículo se fijaron en Bouin para ser procesadas mediante la técnica estándar de inclusión en parafina. Las secciones histológicas (3 μm) se tiñeron con Hematoxilina-eosina y Mallory-Heidenhain-Azan-Gomori's. Para la descripción y registro fotográfico de las células germinales, se utilizó la técnica de microscopía fotónica con el sistema de contraste diferencial de interferencia. Resultados: Los testículos son órganos ovalados, compactos, de color amarillo claro y con poca vascularización. Externamente, están rodeados por una delgada túnica albugínea constituida por tejido conectivo denso regular. Al interior de esta capa se componen por numerosos túbulos seminíferos de contorno hexagonal, en los que se distinguen quistes de células germinativas en diferentes etapas de la espermatogénesis (espermatogonia I y II, espermatocito I y II y espermátidas tempranas y tardías) y espermiogénesis (espermatozoides en fascículos y espermatozoides libres). Separando las estructuras seminíferas se halla el tejido intersticial en el que se destacan las células de Leydig y los vasos sanguíneos. Adicionalmente, en la parte craneal del testículo se encontró el órgano de bidder formado por dos regiones diferenciables, la corteza y la medula. En la corteza se aprecian ovocitos previtelogénicos en diferente tamaño rodeados por una monocapa de células foliculares planas. Por su parte, la región medular es el tejido conectivo que nutre los ovocitos y está constituido por capilares sanguíneos. Conclusiones: Las gónadas de las tres especies analizadas presentan una organización celular quística de manera similar con otros anuros, donde se identificó todos los estadios de la espermatogénesis y espermiogénesis indicando posiblemente una actividad reproductiva continua. Así mismo, se reporta por primera vez el órgano de bidder en las tres especies de Atelopus, lo cual permite sugerir una posible reversión sexual en caso de una disminución poblacional de las hembras como una estrategia reproductiva.
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Animals , Ranidae/anatomy & histology , TestisABSTRACT
Objective:To observe the effect of Wuzi Yanzong Wan made of different processed products on the apoptosis of spermatogenic cells in rats with kidney essence deficiency， and explore its protective effect on spermatogenic cells. Method:SD rats were randomly divided into the blank group， model group， whole raw product group， pharmacopoeia group and salt-processed product group， with 8 rats in each group. The kidney essence deficiency model was replicated by giving tripterygium glycoside tablets （the dose of 20 mg·kg<sup>-1</sup>）. The flow cytometry （FCM） was used to analysis the apoptosis of spermatogenic cells in testis， the immunohistochemistry （IHC） and Western blot were used to detect the expression levels of B-cell lymphoma-2 （Bcl-2） and Bcl-2 associated X protein （Bax） in the testis. High performance liquid chromatography （HPLC） was used to compare the contents of eight components （chlorogenic acid， ellagic acid， hyperoside， isoquercitrin， verbascoside， astragalin， kaempferol and schisandrin） in Wuzi Yanzong Wan made of different processed products， the mobile phase was composed of acetonitrile （A）-0.4% phosphoric acid aqueous solution （B） for gradient elution （0-5 min， 5%-15%A； 5-10 min， 15%-17%A； 10-25 min， 17%A； 25-35 min， 17%-26%A； 35-60 min， 26%-56%A）， the detection wavelength was set at 254 nm. Result:Compared with the model group， the total apoptosis rate of spermatogenic cells， protein expression of Bax and Bcl-2 in each administration group were improved. Among them， the pharmacopoeia group and salt-processed product group had significant effects （<italic>P</italic><0.01）， and the improvement effect of the pharmacopoeia group and salt-processed product group was significantly better than that of the whole raw product group （<italic>P</italic><0.05）. The contents of chlorogenic acid， hyperoside， isoquercitrin and verbascoside in Wuzi Yanzong Wan were increased after the herbal medicines being processed with salt-water. The content of ellagic acid in the salt-processed product group increased， while it decreased in the pharmacopoeia group. The contents of verbascoside， astragalin， kaempferol and schisandrin in samples from the salt-processed product group were greater than those in samples from the pharmacopoeia group. Conclusion:Wuzi Yanzong Wan may reduce the apoptosis of spermatogenic cells in rat testis by inhibiting the expression of Bax and promoting the expression of Bcl-2， and exert its effect of nourishing kidney and enriching essence. The enhanced anti-spermatogenic effect of Wuzi Yanzong Wan after processing may be related to the changes in chemical composition content after processing.
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Objective To observe the effects of high fat dietinduced elevation of blood glucose on the microvascular function of testis and male reproduction in C57BL/6 mice. Methods A total of 40 male C57BL/6 mice were randomly divided into control group and high fat diet (HFD) group (n =20). The mice in HFD group were fed with high fat diet for 20 weeks. Blood glucose and body weight were measured weekly. The permeability of bloodtestis barrier was evaluated by intraperitoneal injection of Evans blue. The blood flow of testicular microcircu-lation and the frequency and amplitude of microvascular vasomotion were detected by laser Doppler blood flow ima-ging system. The morphology of testicular tissue was observed by HE staining. The expressions of platelet- endothelial cell adhesion molecule-1 (PECAM-1/CD31) in testicular microvascular endothelial cells and prolifera-ting cell nuclear antigen (PCNA) in spermatogenic cells were detected by immunohistochemistry. The apoptosis of spermatogenic cells was observed by TUNEL staining. Results The body weight and blood glucose of HFD group were significantly higher than those in control group (P＜0.01). Evans blue staining showed that the integrity of blood-testis barrier of HFD group was damaged, and increased permeability was observed in seminiferous tubules. In HFD group, the mean blood flow of testis and the frequency and amplitude of microvascular vasomotion were sig-nificantly lower than those in control group (P＜0.01). The number of spermatogenic epithelial cells and the thickness of seminiferous epithelium decreased. The expressions of CD31 in microvascular endothelial cells and PCNA in spermatogenic cells were significantly lower in HFD group than those in control group (P＜0.01). The apoptosis level of spermatogenic cells was higher than that in the control group (P <0.01). Conclusions Increased blood glucose level induced by high fat diet in mice can impair the testicular microvasculature and damage the integrity of blood-testis barrier and injure the structure of seminiferous epithelium in mice.
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In artificial insemination, poor quality of semen unsuitable for cryopreservation and susceptibility of spermatozoa to cryodamage in crossbred bulls have been a matter of concern. Present study was designed to identify the testicular cytology indices that might be used to predict the semen quality and cryotolerance of spermatozoa in bulls. Based on the ejaculate rejection rate and sperm cryotolerance, bulls (Holstein Friesian X Tharparkar crossbred) were classified into either good (producing good quality semen with spermatozoa having good cryotolerance; n=4) or poor (producing poor quality semen with spermatozoa having poor cryotolerance; n=4). Testicular cytology was studied in all the 8 bulls using fine needle aspiration technique. Testicular cytology of good bulls and poor bulls differed significantly. The proportion of Sertoli cells was significantly higher in good bulls (25.3±1.6) compared to poor bulls (11.0±0.8). The Sertoli cell index was 46.1±5.0 in good bulls while it was only 13.8±1.3 in poor bulls. The cut off values, as determined using Receiver Operating Characteristics analysis, indicate that the bulls having testicular cytogram comprising of <15.5% Sertoli cells, <24.3 Sertoli cell index and >4.0 spermatogenic cells to Sertoli cell ratio might be a poor bull in terms of semen quality and cryotolerance of spermatozoa. The proportion of Sertoli cells in the testicular cytology had positive (P <0.05) relationship with semen quality and cryotolerance of spermatozoa.
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Background: Caverta (Sildenafil citrate), an oral therapy for Erectile Dysfunction (ED), being the citrate salt of Sildenafil, is a selective inhibitor of cGMP- specific phosphodiestrase type (PDE5). Aim: To study the drug- induced (i) changes in the trace element content of Testis and (ii) changes in the histoarchitecture of Testis of the experimental Albino rats. Materials and Methods: For the present study, totally 48 animals were selected on weight basis and divided into 8 groups (S1, S2, S3, S4, S5, S6, S7 and S8) with six animals in each group. Control animals (S1) were fed with conductivity water while the experimental animals (S2, S3, S4 and S5) were treated with a single dose of Caverta (@ 1μg/g body weight). Control animals were sacrificed at zero hour while the experimental animals (S2, S3, S4 and S5) were sacrificed after 1 hour, 2½ hours, 4 hours and 24 hours of drug administration respectively S6, S7 and S8 group of animals were fed with a single dose of the chosen drug (@ 1μg/g body weight) daily for all the 15, 30 and 45 days respectively. These animals were sacrificed after 4 hours of the last dosage. Vertical ventral midline incision was made in the abdominal wall to collect the left and right Testes. Results: The spectral analysis indicates that the long term Caverta treatment of Albino rats results in the accumulation of Iron and Copper levels accompanied by a depletion of Nickel levels in the Testis. The histological studies indicate that long term exposure of Testis to Caverta leads to distorted histoarchitecture of the seminiferous tubules, interstitial space dilation and separation of Spermatogenic cells. Conclusion: Long term Sildenafil Citrate (Caverta) treatment of Albino rats will bring in adverse effects and will completely alter the histoarchitecture of the Testis.
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<p><b>OBJECTIVE</b>To investigate the effect of simulated microgravity and carbon ion irradiation (CIR) on spermatogenic cell apoptosis and sperm DNA damage to the testis of male Swiss Webster mice, and assess the risk associated with space environment.</p><p><b>METHODS</b>Sperm DNA damage indicated by DNA fragmentation index (DFI) and high DNA stainability (HDS) was measured by sperm chromatin structure assay (SCSA). Apoptosis of spermatogenic cells was detected by annexin V-propidium iodide assay. Bax (the expression levels of p53) and proliferating cell nuclear antigen (PCNA) were measured by immunoblotting; p53 and PCNA were located by immunohistology.</p><p><b>RESULTS</b>HDS, DFI, apoptosis index, and the expression levels of p53 and Bax were detected to be significantly higher in the experimental groups (P<0.05) compared with those in the control group; however, the PCNA expression varied to a certain degree. p53- and PCNA- positive expression were detected in each group, mainly in relation to the spermatogonic cells and spermatocytes.</p><p><b>CONCLUSION</b>The findings of the present study demonstrated that simulated microgravity and CIR can induce spermatogenic cell apoptosis and sperm DNA damage. Sperm DNA damage may be one of the underlying mechanisms behind male fertility decline under space environment. These findings may provide a scientific basis for protecting astronauts and space traveler's health and safety.</p>
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Animals , Male , Mice , Apoptosis , Radiation Effects , Carbon , Cell Proliferation , Radiation Effects , DNA Damage , Heavy Ions , Immunohistochemistry , Random Allocation , Sperm Count , Spermatogenesis , Radiation Effects , Spermatozoa , Radiation Effects , Testis , Radiation Effects , Weightlessness SimulationABSTRACT
Reproductive aspects of the Brazilian snapper Lutjanus alexandrei, were characterized, including a description of the development of oocytes and spermatogenic cells, size at first sexual maturity, and fecundity. A total of 540 fish were analyzed with 250 having their gonads sectioned to allow microscopic evaluation. Six maturity stages were identified for females and males: immature, maturing, mature, spawning, spawned, and resting. Fish standard length (SL) varied from 13.0 to 28.3 cm and sex ratio was 1.6 males: 1.0 females. Monthly distributions of mean Gonadosomatic Index (GSI) and maturity stages suggest that spawning occurs mainly in a protracted period, during the warmer months, from November to March. The size of first sexual maturity was estimated at 17.1 cm SL for females and 16.8 cm SL for males. Oocyte development suggests that L. alexandrei exhibits a multiple batch spawning behavior and batch fecundity varied from 34,000 to 324,000 oocytes.
Os aspectos reprodutivos da baúna-de-fogo Lutjanus alexandrei foram caracterizados, incluindo a descrição do desenvolvimento dos ovócitos e células espermatogênicas, do tamanho de primeira maturação sexual, e da fecundidade. Um total de 540 peixes foi analisado, dos quais 250 tiveram as suas gônadas seccionadas para avaliação microscópica. Seis estágios de maturidade sexual foram determinados para fêmeas e machos: imaturo, em maturação, maduro, desovando, desovado e repouso. O comprimento padrão (CP) dos peixes variou de 13,0 a 28,3 cm e a proporção sexual foi de 1,6 machos: 1,0 fêmeas. As distribuições mensais dos valores médios do Índice Gonadosomático (IGS) e dos estágios de maturidade sexual sugerem a ocorrência de desovas em um período prolongado, principalmente nos meses de temperaturas mais quentes, entre novembro e março. O tamanho médio de primeira maturação sexual foi estimado em 17,1 cm CP para as fêmeas e 16,9 cm CP para os machos. O padrão de desenvolvimento dos ovócitos sugere que L. alexandrei exibe comportamento de múltiplas desovas por lote, e a fecundidade variou entre 34.000 a 324.000 ovócitos.
Subject(s)
Animals , Oocytes/growth & development , Perciformes/growth & development , Reproduction/physiology , Semen/physiology , Embryonic Development/geneticsABSTRACT
Objective To investigate potential protection by protect against ethanol-induced apoptosis of spermatogenic cells in rat testis.Methods Thirty SD adult male rats were randomly divided into three groups: normal group,alcohol group and puerarin group.At 40th day,BCL-2 and BAX of spermatogenic cells of testis tissue were checked by RT-PCR and immunohistochemistry;Apoptosis of spermatogenic cells was determined by TUNEL.Results The results of RT-PCR and immunohistochemistry indicated that BCL-2 and BAX of spermatogenic cells were not significanty different between puerarin group and normal group,but there was the significant difference between alcohol group and puerarin group(P
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0.05).Except that the wet weight of the epididymis in high-dose group of apigenin was significantly different compared with middle-dose group of apigenin(P 0.05).Cell cycle analysis showed that compared with the negative control group,the percent of diploid cells in low and high-dose group was decreased,the proportion of G0/G1 phase cells in low-dose group of apigenin was reduced(P
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Mesna has been used with ifosfamide to prevent urotoxicity in the treatment of testicular cancers. This drug also protected the toxicities of adriamycin without compromising cytostatic activity. With n idea of radioprotective role of sulfhydryl group of radioprotectors and of mesna decreasing the toxic effect of adriamycin which produces free radicals, mesna and radiation were administered to mice to study the protective effect of this drug and to identify the difference in regenerative capacity of the germ cells in the testis between radiation-treated and both mesna- and radiation-treated groups. The shape and numbers of spermatogenic cells in the seminiferous tubules were examined every week after irradiation. In both groups, initial reduction and later recovery in germ seel numbers and shape was observed. The lowest germ cell number was found around three weeks after irradiation. Mean germ cell number of the mesna-treated group was significantly higher than radiation-treated group at all observed periods (p<0.05). More competent regeneration was present in mesna-treated group. These results suggest that mesna protect the testis from radiation injury. Further study will be necessary to identify whether mesna protects other tissues from radiation and it does not hamper tumor control.
Subject(s)
Animals , Mice , Doxorubicin , Free Radicals , Germ Cells , Ifosfamide , Mesna , Radiation Injuries , Regeneration , Robenidine , Seminiferous Tubules , Testicular Neoplasms , TestisABSTRACT
Objective To inquire into the relationship between infections of Toxoplasma gondii and apoptosis of spermatogenic cells. Methods Apoptotic spermatogenic cells of mice infected with Toxoplasma gondii were examined by Wright-Giemsa staining and terminal deoxynucleotidyl trans-Icrase (TdT) mediated deoxyuridine triphosphate (dUTP)-biotin nick-end labeling (TUNED techniques. Results Apoptosis rate of infective group of Toxoplasma was significantly higher than thai oi control group (F
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Objective To prove the mechanism of diethylstibestrol(DES) induced abnormal spermatogenesis,this study on the changes of proto-oncogenes bcl-2 and p53 proteins as well as cadherin adherens molecules expression in adult male hamster spermatogenic cells. Methods Adult male golden hamsters were treated daily olive oil vehicle either alone(control)or containing 1 mg/kg body weight of DES (experiment)by subcutaneous injection for 7 days.The changes of testes histological morphology were observed under light and electron microscope.The expressions of bcl-2 and p53 and cadherin in the testes seminiferous epithelium were detected by immunohistochemistry. Results DES induced adult hamster testes abnormal spermatogenesis.As compared to the control group animals,in DES-treated group animals the expression of both bcl-2 and p53 significantly increased in spermatogenic cells,especially in spermatocytes and round spermatids.However,the expression of cadherin adherens molecules obviously decreased.Conclusion It is one of reasons of DES inducing abnormal spermatogenesis that DES can result in increasing in the expression of proto-oncogenes bcl-2 and p53,and reducing cadherin adherens molecules expression.
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Objective To evaluate the effect of sodium nitroprusside(SNP)and N-nitro-l-arginine-mythel-ester (L-NAME) on apoptosis of spermatogenic cells in rats. Methods Fourty adult male Sprague-Dawley rats (60-70days) were divided into four groups.Each group was injected intraperitoneally with one of the following agents, once a day, for 12 days: 1. SNP; 2.L-NAME;3.SNL+L-NAME;4.Normal saline NS group.Two hours after the last time injection the rats were sacrificed.TUNEL staining and flow cytometry analysis were used to detect the apoptosis of spermatogenic cells. Results Sub-monoploid and apoptosis index (AI) in SNP group was significantly higher than that of NS group and sub-monoploid and apoptosis index (AI) in L-NAME group were significantly lower than that of NS group by FCM and TUNEL (P0.5) was found.Conclusion SNP can accelerate the apoptosis of spermatogenic cells and L-NAME can inhibite the apoptosis of spermatogenic cells,The effect of SNP and L-NAME on apoptosis of spermatogenic cells probably occurs through the action of nitric oxide.