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1.
Säo Paulo med. j ; 140(2): 163-170, Jan.-Feb. 2022. tab
Article in English | LILACS | ID: biblio-1366043

ABSTRACT

Abstract BACKGROUND: Because normal male sexual differentiation is more complex than normal female sexual differentiation, there are more cases of disorders of sex development (DSDs) with 46,XY karyotype that have unclear etiology. However, Leydig and Sertoli cell markers are rarely used in distinguishing such individuals. OBJECTIVES: To evaluate the function of Leydig and Sertoli cells in individuals with genital ambiguity, 46,XY karyotype, palpable gonads and normal testosterone secretion. STUDY DESIGN AND SETTING: Case-control study with 77 patients, including eight with partial androgen insensitivity syndrome, eight with 5α-reductase deficiency type 2 (5ARD2) and 19 with idiopathic 46,XY DSD, and 42 healthy controls, from the Interdisciplinary Study Group for Sex Determination and Differentiation (GIEDDS), at the State University of Campinas (UNICAMP), Campinas, Brazil. METHODS: Baseline levels of gonadotropins, anti-Müllerian hormone (AMH), inhibin B, insulin-like 3 (INSL3), testosterone and dihydrotestosterone in cases, and AMH, inhibin B, and INSL3 levels in controls, were assessed. RESULTS: There was no significant difference in age between cases and controls (P = 0.595). AMH and inhibin B levels were significantly lower in cases than in controls (P = 0.031 and P < 0.001, respectively). INSL3 levels were significantly higher in cases than in controls (P = 0.003). Inhibin B levels were lower in 5ARD2 patients (P = 0.045) and idiopathic patients (P = 0.001), in separate comparisons with the controls. CONCLUSION: According to our findings, we can speculate that inhibin B levels may be used to differentiate among DSD cases.


Subject(s)
Humans , Male , Female , Sertoli Cells/metabolism , Disorders of Sex Development , Testosterone/metabolism , Case-Control Studies , Karyotype , Gonads/metabolism
2.
Int. braz. j. urol ; 45(4): 815-824, July-Aug. 2019. tab, graf
Article in English | LILACS | ID: biblio-1019894

ABSTRACT

ABSTRACT Introduction Chronic hyperglycemia is caused by diabetes mellitus-committed genital morphophysiology, and oxidative stress is one of the main factors involved in this process. Alpha lipoic acid (ALA) can prevent metabolic and morphological changes in diabetic individuals. Objectives In present study, we evaluated the effects of regular ALA consumption on the spermatogenesis and histoarchitecture in the male genital system of diabetic rats. Materials and Methods Thirty-two Wistar rats were divided into groups: Control (CG); Diabetic Control (DCG), receiving commercial diet: ALA Group (ALAG) and Diabetic ALA Group (DALAG), fed diets with added ALA (300 mg/Kg bw). The diabetic groups received a single injection of streptozotocin (60 mg/kg). After sixty days of the diet, the animals were euthanized, and semen, testis and epididymis samples were collected. A histomorphometric analysis was performed to determine the epithelial height, tubular and luminal diameter, tubular and luminal area of seminiferous tubules and each epididymal region. Sertoli cells were evidenced using the antivimenti antibody and were quantified. The results were statistically analyzed by the ANOVA test. Results At the end of the experiment, the DALAG glycemia was significantly lower than DCG. The histomorphometric parameters of the seminiferous and epididymal tubules did not show improvement in the DALAG. However, there was an improvement in the DALAG in terms of the concentration, motility and percentage of spermatic pathologies, as well as in the number of Sertoli cells (p<0.001). Conclusions The results demonstrated that supplementation with the ALA antioxidant retards testicular lesions and preserve the process of spermatogenesis in diabetes.


Subject(s)
Animals , Male , Spermatozoa/drug effects , Testis/drug effects , Thioctic Acid/pharmacology , Diabetes Mellitus, Experimental/pathology , Epididymis/drug effects , Antioxidants/pharmacology , Sertoli Cells , Sperm Count , Sperm Motility/drug effects , Spermatogenesis/drug effects , Spermatogenesis/physiology , Spermatozoa/physiology , Testis/physiopathology , Testis/pathology , Immunohistochemistry , Random Allocation , Reproducibility of Results , Rats, Wistar , Diabetes Mellitus, Experimental/physiopathology , Epididymis/pathology
3.
Article in English | WPRIM | ID: wpr-742360

ABSTRACT

Male infertility (MI) is a complex multifactorial disease, and idiopathic infertility accounts for 30% of cases of MI. At present, the evidence for the effectiveness of empirical drugs is limited, and in vitro fertilization is costly and may increase the risk of birth defects and childhood cancers. Therefore, affected individuals may feel obliged to pursue natural remedies. Traditional Chinese medicine (TCM) may represent a useful option for infertile men. It has been demonstrated that TCM can regulate the hypothalamic-pituitary-testicular axis and boost the function of Sertoli cells and Leydig cells. TCM can also alleviate inflammation, prevent oxidative stress, reduce the DNA fragmentation index, and modulate the proliferation and apoptosis of germ cells. Furthermore, TCM can supply trace elements and vitamins, ameliorate the microcirculation of the testis, decrease the levels of serum anti-sperm antibody, and modify epigenetic markers. However, the evidence in favor of TCM is not compelling, which has hindered the development of TCM. This review attempts to elucidate the underlying therapeutic mechanisms of TCM. We also explore the advantages of TCM, differences between TCM and Western medicine, and problems in existing studies. Subsequently, we propose solutions to these problems and present perspectives for the future development of TCM.


Subject(s)
Apoptosis , Congenital Abnormalities , DNA Fragmentation , Epigenomics , Fertilization in Vitro , Germ Cells , Humans , Infertility , Infertility, Male , Inflammation , Leydig Cells , Male , Male , Medicine, Chinese Traditional , Microcirculation , Oxidative Stress , Sertoli Cells , Testis , Trace Elements , Vitamins
4.
Biosci. j. (Online) ; 34(6): 1697-1705, nov.-dec. 2018. ilus
Article in English | LILACS | ID: biblio-968970

ABSTRACT

The hoary fox Lycalopex vetulus (Lund, 1842) is a small canid, endemic to Brazil, belonging to the Canidae family, widely distributed in the country, occurring records in different regions and habitats. The objective of this study is to describe morphologically the testicles and epididymal ducts of hoary fox. The animals, coming from the zoo of Federal University of Mato Grosso, Brazil, had died by natural causes. The male reproductive system was dissected to collect the testicles. The samples were fragmented and histologically examined. Microscopically, the testes were coated by the vaginal and albuginea tunic, formed by modeled dense connective tissue with large amount of collagen fibers. Into the organ, convoluted seminiferous tubules were surrounded by a basement membrane characterized by the presence of myoid and Sertoli cells and germinative epithelium composed by Between the seminiferous tubules, interstitial tissue composed of connective tissue, blood and lymph vessels and Leydig cells in polyhedral shape was present. The epididymal ducts showed pseudostratified columnar epithelium with secretory cells, in which stereocilia located on a basement membrane filled by myoid cells were found. The structures observed by us are very similar to those described for other mammals.(AU)


A raposa-do-campo Lycalopex vetulus (Lund, 1842) é um canídeo de pequeno porte, endêmico do Brasil, pertencente a família Canidae, com ampla distribuição no país, ocorrendo registros em várias regiões e habitats diferentes. Com base nessa informação, o objetivo deste trabalho é caracterizar morfologicamente os testículos e ductos epididimários da raposa-do-campo. O animal, oriundo do zoológico da Universidade Federal de Mato Grosso, Brasil, veio a óbito por causas naturais e o sistema reprodutor masculino foi dissecado para coleta dos testículos. As amostras retiradas foram fragmentadas e histológicamente examinadas. A partir das análises microscópicas dos testículos foram identificados: a túnica vaginal e albugínea, formada por tecido conjuntivo denso modelado, com grande quantidade de fibras colágenas; túbulos seminíferos enovelados e revestidos por epitélio germinativo e células de Sertoli, envolvidos por uma membrana basal com presença de células mioides; tecido intersticial entre os túbulos seminíferos constituído de tecido conjuntivo, vasos sanguíneos e linfáticos, e células de Leydig em formato poliédrico. Os ductos epididimários apresentaram epitélio cilíndrico pseudoestratificado com células secretoras dos quais projetam estereocílios, situados sobre uma membrana basal repleta por células mióides. As estruturas por nós observadas possuem muita semelhança com as demais descrições para mamíferos.(AU)


Subject(s)
Testis , Foxes , Genitalia, Male , Seminiferous Tubules , Sertoli Cells , Basement Membrane , Connective Tissue , Canidae , Stereocilia , Leydig Cells
5.
Rev. Hosp. Niños B.Aires ; 60(270): 214-222, sept. 2018.
Article in Spanish | LILACS | ID: biblio-998451

ABSTRACT

Introducción: La capacidad funcional del testículo en los niños con criptorquidia ha recibido poca atención. La hormona anti-mülleriana (AMH), producida por la célula de Sertoli, es el marcador ideal para evaluar la función testicular durante la infancia. Objetivo: Caracterizar la función testicular en niños prepuberales antes de la orquidopexia. Investigar la asociación entre función testicular y las características de la criptorquidia. Pacientes y métodos: Estudio de corte transversal y analítico, retrospectivo. Medida de resultado principal: concentración de AMH. Medidas de resultados secundarias: concentraciones de gonadotrofinas y testosterona. Para comparación, se utilizaron los niveles hormonales de 179 niños normales. Resultados: Se seleccionaron 186 pacientes con criptorquidia bilateral y 124 con criptorquidia unilateral. La mediana de SDS de AMH fue menor a 0 en ambos grupos. La concentración sérica de AMH fue más baja en pacientes con criptorquidia bilateral que en niños controles y en niños con criptorquidia unilateral. La testosterona estuvo disminuida en niños menores de 6 meses. Las gonadotrofinas estuvieron aumentadas en un bajo porcentaje de los casos. Conclusión: Los niños prepuberales con criptorquidia, especialmente aquellos con criptorquidia bilateral, tienen menor producción de AMH y una considerable prevalencia de disfunción testicular


Introduction: Little information is available on testicular function in boys with cryptorchidism. Anti-müllerian hormone (AMH) is a good marker of testicular functionin childhood. Objective: the aim of this study was to assess testicular function in boys with cryptorchidism before orchiopexy, and to look for an association between testicular function and features of cryptorchidism. Patients and methods: We performed a cross-sectional, retrospective study. Main outcome measure was serum AMH concentration, and secondary variables were gonadotropin and testosterone concentrations. For comparison, levels in 179 normal boys were compared. Results: 186 boys with bilateral cryptorchidism and 124 with unilateral cryptorchidism were included. Mean SDS AMH was below 0 in both groups. Mean serum AMH was lower in boys with bilateral cryptorchidism, as compared to unilateral cryptorchidism and controls between 6 months and 8.9 years of age. Testosterone was lower than normal in boys < 6 months of age. Gonadotropins were rarely affected. Conclusions: Prepubertal boys with cryptorchidism, especially those with bilateral forms, have a lower AMH production, reflecting testicular dysfunction


Subject(s)
Male , Cryptorchidism , Gonadotropins , Hypogonadism , Pediatrics , Sertoli Cells , Testosterone
6.
Biol. Res ; 51: 12, 2018. graf
Article in English | LILACS | ID: biblio-950898

ABSTRACT

BACKGROUND: Chemokine (C-C motif) receptor 6 (CCR6) is present in sperm and plays a significant role in sperm motility and chemotaxis acting in the reproductive tracts. However, the expression and functional significance of CCR6 in testis are still poorly understood, especially in the process of spermatogenesis. METHODS AND RESULTS: CCR6 was expressed in spermatogenic cell lines and its expression was shown in an age-dependent upregulation manner from puberty to adulthood in mouse testis. Immunostaining results confirmed the localization of CCR 6 in testis. Further chemotaxis assays demonstrated that spermatogenic cells GC-1 and -2 exhibited a directional movement toward CCR6-specific ligand such as CCL20 or Sertoli cells in vitro. CONCLUSIONS: The present findings indicate that CCR6 is involved in the chemotaxis of spermatogenic cells in vitro and promotes chemotaxis under non-inflammatory conditions during normal spermatogenesis.


Subject(s)
Humans , Animals , Male , Mice , Rabbits , Spermatogenesis/physiology , Chemotaxis/physiology , Cryptorchidism/metabolism , Chemokine CCL20/metabolism , Receptors, CCR6/metabolism , Sertoli Cells , Sperm Motility/physiology , Testis/physiology , Immunohistochemistry , Blotting, Western , Fluorescent Antibody Technique , Mice, Inbred C57BL
7.
Anatomy & Cell Biology ; : 52-61, 2018.
Article in English | WPRIM | ID: wpr-713349

ABSTRACT

Cimetidine is an H2 receptor antagonist that has an antiandrogenic effect. It intervenes with the conversion of testosterone into estrogen in the Sertoli cells with accompanying testicular structural changes. In the present study, the microscopic and the ultrastructural changes induced by cimetidine and the effect of vitamin B12 as a protective agent on rat testes were studied. Immunoexpression of estrogen receptor β (ERβ) in testes was evaluated. Twenty-four adult male rats were divided into four groups: control, cimetidine-treated, vitamin B12 treated, and combined cimetidine and vitamin B12 treated. The experimental rats were administered with cimetidine and/or vitamin B12 for 52 days. Group II rats showed marked atrophy of the seminiferous tubules with a significant increase in tubular diameter and decrease in the tubular luminal and epithelial areas. Ultrastructure of this group showed irregular Sertoli cells with basal cytoplasmic vacuolation and significantly thickened basement membrane. ERβ immunoexpression was similar to controls. Group III rats showed near normal seminiferous tubular structures with minimal cellular alterations and the immunoreactivity of the testicular sections was very close to normal. However, group IV rats showed markedly immunopositive detached cells, spermatids, and primary spermatocytes. Cimetidine interferes with the control of spermatogenesis as evidenced by microscopic and ultrastructural studies and affection of ERβ receptors and vitamin B12 has a protective action against this harmful effect.


Subject(s)
Adult , Animals , Basement Membrane , Cimetidine , Cytoplasm , Estrogens , Humans , Male , Phenobarbital , Rats , Seminiferous Epithelium , Seminiferous Tubules , Sertoli Cells , Spermatids , Spermatocytes , Spermatogenesis , Testis , Testosterone , Vitamin B 12 , Vitamins
8.
National Journal of Andrology ; (12): 221-225, 2018.
Article in Chinese | WPRIM | ID: wpr-689773

ABSTRACT

<p><b>Objective</b>To study the expression of CLAUDIN-11 in the testis tissue of non-obstructive azoospermia (NOA) patients with different severities and investigate its clinical significance.</p><p><b>METHODS</b>Sixty-two NOA patients were divided into a hypospermatogenesis (HS) group (n = 30) and a Sertoli cell only syndrome (SCO) group (n =32). The expression of CLAUDIN-11 in the testicular tissue of the patients was detected by immunohistochemistry, that of CLAUDIN-11 mRNA determined by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the levels of serum reproductive hormones measured by chemiluminescent immunoassay.</p><p><b>RESULTS</b>Immunohistochemistry showed that the expression of CLAUDIN-11 was mainly in the cytoplasm of the Sertoli cells around the seminiferous tubule wall in the HS group, but diffusely distributed in the membrane of the Sertoli cells in the SCO group. RT-qPCR revealed a significantly lower expression of CLAUDIN-11 mRNA in the HS than in the SCO group (0.008 ± 0.001 vs 0.013 ± 0.002, t = 10.616, P<0.01). The level of serum luteotropic hormone (LH) was also markedly lower in the HS than in the SCO group ([3.62 ± 1.34] vs [4.96 ± 3.10] IU/L, P<0.05) and so was that of follicle-stimulating hormone (FSH) ([5.36 ± 2.80] vs [10.65 ± 9.18] IU/L, P<0.05).</p><p><b>CONCLUSIONS</b>The up-regulated expression of CLAUDIN-11 in Sertoli cells may play an important role in the development and progression of spermatogenic dysfunction in NOA patients.</p>


Subject(s)
Azoospermia , Genetics , Metabolism , Claudins , Metabolism , Follicle Stimulating Hormone , Metabolism , Humans , Male , Oligospermia , Genetics , Metabolism , RNA, Messenger , Metabolism , Seminiferous Tubules , Metabolism , Sertoli Cell-Only Syndrome , Genetics , Metabolism , Sertoli Cells , Metabolism , Spermatogenesis , Testis , Metabolism
9.
Article in Chinese | WPRIM | ID: wpr-941670

ABSTRACT

OBJECTIVE@#To investigate the effect of triptolide (TP) on oxidative stress and apoptosis in TM4 sertoli cells and related molecular mechanism.@*METHODS@#TM4 cells were incubated with different concentrations of triptolide for 24 h, then collected for further experiments. Cell proliferation analysis was used to measure the inhibitive effect of triptolide on proliferation of TM4 cells; DCFH-DA (6-carboxy-2',7'-dichlorofluorescein diacetate) probe was used to stain the TM4 cells, the level change of intracellular ROS was discovered through flow cytometry; the TM4 cells were stained by Annexin V-FITC/PI to detect whether triptolide induced apoptosis in the TM4 cells; Protein was extracted from the TM4 cells in control and triptolide group. Western blot was performed to determine the expression of apoptosis marker protein cleaved-PARP and PI3K/Akt signaling pathway-related proteins [p-Akt (Ser473), Akt, p-mTOR (Ser2448), mTOR, p-p70S6K (Thr389), p70S6K].@*RESULTS@#Cell proliferation analysis revealed that triptolide reduced the TM4 cells viability significantly compared with control group in a dosage-dependent manner [10 nmol/L: (73.77±20.95)%, 100 nmol/L: (51.60±10.43)%, 500 nmol/L: (44.34±5.78)%]; The level of intracellular ROS in the TM4 cells was significantly induced in a dosage-dependent manner (P<0.01); triptolide remarkably induced early-stage and late-stage apoptosis in the TM4 cells [control: (3.84±1.50)%, 100 nmol/L: (13.04±2.03)%, 200 nmol/L: (16.24±1.34)%, 400 nmol/L: (18.76±3.45)%]; The expression of cleaved-PARP was significantly upregulated in the TM4 cells after incubation with triptolide (P<0.01); The expression levels of p-Akt/Akt and p-p70S6K/p70s6k were significantly increased compared with control group (P<0.01). No significant change was observed among the expression levels of p-mTOR/mTOR (P>0.05).@*CONCLUSION@#In vitro studies showed that triptolide could effectively suppress the proliferation and induce apoptosis of TM4 sertoli cells. The oxidative stress was upregulated after incubation with triptolide, which may be one of the mechanisms of cytotoxicity in TM4 cells. Treatment of triptolide led to activation of Akt and p70S6K, indicating that the PI3K/Akt signaling pathway may be involved in response to oxidative stress in TM4 cells. The activation of PI3K/Akt signaling pathway was one of the molecular mechanisms involved in triptolide-mediated oxidative stress in TM4 cells. Our study provides insight into alleviating reproductive toxicity of triptolide in clinical and developing male contraceptive.


Subject(s)
Antineoplastic Agents, Alkylating/pharmacology , Apoptosis/drug effects , Diterpenes/pharmacology , Epoxy Compounds/pharmacology , Humans , Male , Oxidative Stress/drug effects , Phenanthrenes/pharmacology , Phosphatidylinositol 3-Kinases , Proto-Oncogene Proteins c-akt/drug effects , Sertoli Cells/drug effects , Signal Transduction/drug effects
10.
Egyptian Journal of Hospital Medicine [The]. 2018; 71 (4): 2892-2906
in English | IMEMR | ID: emr-192544

ABSTRACT

Background: The reproductive ability of sexually mature males is dependent on the capacity of the testes tubules to produce large numbers of highly viable sperms and the interstitial cells to produce adequate levels of androgens to ensure maturation of the reproductive system. Azoospermia, defined as the complete absence of spermatozoa in the ejaculate, invariably results in infertility but does not necessarily imply sterility. It is identified in approximately 1% of all men and in 10 to 15% of infertile males


Patients and methods: The use of biopsies had been approved by the ethics committee of the Faculty of Medicine, Al Azhar University. All testicular biopsies were taken after written informed consent at the Department of Clinical Andrology, International Islamic Center for Population and Research Studies. Eleven patients with idiopathic infertility, were enrolled in this study and were assessed and diagnosed by a complete examination and specific investigations


Results: This study included 11 patients. They were divided into two groups obstructive azoospermia [OA] and Non-obstructive azoospermia. Non-obstructive azoospermia is subdivided into 4 groups: Early maturation arrest, late maturation arrest, Sertoli cell only [Sco] and Klienfilter syndrome


Conclusion: Spermatogenesis and maintenance of reproductive functions are controlled by androgens which are steroid hormones. Androgens exert most of their effects through genomic actions, which involve their binding to the androgen receptor [AR].AR plays important roles during later stages of sperm formation and maturation by influencing the four major cell types: Sertoli, Leydig, peritubularmyoid, and germ cells. Except for maturation arrest non-obstructive azoospermia, AR positive staining was not detected in germ cells. The highest number of Sertoli cell androgen receptors was in sections of testis from obstructive azoospermia and non- obstructive late maturation arrest patients, Testicular sections from patients with Klinefelter syndrome presented the lowest number of androgen receptor positive cells. It was concluded that the present study revealed that demonstration of androgen receptors in testicular sections is a good indicator of spermatogenic activity. Such information is valuable for the decision of obtaining spermatozoa from the testis for ICSI


Subject(s)
Humans , Male , Adult , Microscopy , Azoospermia , Receptors, Androgen , Sertoli Cells , Infertility, Male , Spermatogenesis
11.
National Journal of Andrology ; (12): 168-171, 2018.
Article in Chinese | WPRIM | ID: wpr-775200

ABSTRACT

Inhibin B, a glycoprotein produced predominantly by Sertoli cells and preferentially suppressing the production and secretion of follicle-stimulating hormone (FSH) in the pituitary, is closely related to spermatogenesis. Varicocele is the abnormal dilatation and tortuosity of the pampiniform plexus veins, which may contribute to spermatogenic dysfunction and male infertility. More and more evidence has shown that the level of serum inhibin B is negatively correlated with the severity of varicocele. Determination of the inhibin B level may help assess the severity of spermatogenic dysfunction of the patient and predict the outcomes of varicocele repair and therefore has a potential application value in the diagnosis and treatment of varicocele.


Subject(s)
Follicle Stimulating Hormone , Metabolism , Humans , Infertility, Male , Blood , Inhibins , Blood , Male , Sertoli Cells , Spermatogenesis , Varicocele , Blood
12.
Int. j. morphol ; 35(4): 1597-1606, Dec. 2017. tab, graf
Article in Spanish | LILACS | ID: biblio-893174

ABSTRACT

RESUMEN: Las células madre de la línea germinal masculina son factores clave para la espermatogénesis masculina y la fertilidad. Las células sustentaculares (células de Sertoli) como células somáticas juegan un papel fundamental en la creación de un microambiente esencial para la auto-renovación y diferenciación de las células de la línea germinal masculina. Las células madre mesenquimales son reconocidas como células auto-renovables y multipotentes capaces de diferenciarse en múltiples tipos de células. La generación de células germinales masculinas a partir de células madre mesenquimales puede proporcionar un método terapéutico para tratar la infertilidad masculina. En este estudio, las células mesenquimales derivadas de la médula ósea (BMMSCs) se recuperaron de la médula ósea de ratones de 6-8 semanas de edad del Instituto de Investigación Médico Naval (NMRI). En el estudio se aislaron las células sustentaculares y se enrriquecieron usando placas revestidas con lectina. Se obtuvo el medio de condición celular después de diferentes intervalos de tiempo. Posteriormente se cultivaron las BMMSC con diferentes concentraciones de SCCM y medio de Eagle modificado por Dulbecco (DMEM) en diversos momentos. Se evaluaron marcadores específicos de células de línea germinal usando la reacción en cadena de polimerasa transcriptasa inversa (RT-PCR) e inmunocitoquímica. Los resultados mostraron que las BMMSCs cultivadas con SCCM durante 48h exhibieron transcritos específicos de línea germinal (Mvh, Iid4, piwil2) (p <0,05) y marcadores (Mvh, Scp3). Nuestros resultados indican que el cultivo de BMMSCs con SCCM puede conducir a la diferenciación efectiva de BMMSCs en células germinales y proporcionar una estrategia de tratamiento para la infertilidad masculina.


SUMMARY: Male germ line stem cells are key factors for male spermatogenesis and fertility. Sustentacular cells (Sertoli cells) as somatic cells play a pivotal role in creating essential microenvironment for the self-renewal and differentiation of the male germ line cells. Mesenchymal stem cells are recognized as self-renewing and multipotent cells able to differentiate into multiple cell types. The generation of male germ cells from mesenchymal stem cells may provide a therapeutic method to treat male infertility. In this study, Bone marrow derived mesenchymal cells (BMMSCs) were retrieved from the bone marrow of 6-8-week old Naval Medical Research Institute (NMRI) mice. Sustentacular cells (Sertoli cells) were isolated and made rich using lectin coated plates. Sustentacular cell condition medium (SCCM) was collected after different time intervals. Then the BMMSCs were cultured with different concentration of SCCM and Dulbecco's Modified Eagle's medium (DMEM) at various times. Specific markers of Germ line cells were evaluated by using Reverse transcriptase polymerase chain reaction (RT-PCR) and immunocytochemistry. The results showed that BMMSCs cultured with SCCM for 48h exhibited germ line specific transcripts (Mvh, Iid4, piwil2) (p< 0.05) and markers (Mvh, Scp3). Our findings represent that culturing BMMSCs with SCCM may lead to effective differentiation of BMMSCs into germline cells and provide a treatment strategy for male infertility.


Subject(s)
Animals , Male , Mice , Sertoli Cells/cytology , Mesenchymal Stem Cells/cytology , Sertoli Cells/ultrastructure , Testis/cytology , Bone Marrow , Immunohistochemistry , Cell Differentiation , Culture Media, Conditioned , Reverse Transcriptase Polymerase Chain Reaction , Flow Cytometry
13.
Int. j. morphol ; 35(4): 1444-1450, Dec. 2017. graf
Article in Spanish | LILACS | ID: biblio-893155

ABSTRACT

RESUMEN: La espermatogénesis es un proceso continuo que se inicia durante el desarrollo embriofetal. Las relaciones auto, para y yuxtacrinas indican la interdependencia de las células intersticiales (de Leydig) con las células peritubulares (lamina propia) y células sustentaculares (de Sertoli). Ciertos morfógenos son fundamentales en este proceso. Las células sustentaculares son capaces de regular la diferenciación y función de las células peritubulares e intersticiales a través de la producción de IGF1, TGFA, TGFB y DHH. Las células peritubulares son capaces de producir P-Mod-S, regulando la diferenciación de las células sustentaculares, y a través de FGF2 y FGF9 modulan las transiciones epitelio-mesenquimática entre células sustentaculares y mesonefros. También remodelan la membrana basal del condón testicular y regulan la diferenciación y función de las células intersticiales por medio de IGF1, TGFA y TGFB. Las células intersticiales son las reponsables de la producción de testosterona e INSL3, influyendo en la diferenciación sexual masculina. Se plantea que provienen de células mesenquimales del epitelio celómico y mesonefros. Sin embargo, otros autores proponen su origen a partir de células de la cresta neural. Estas influyen a través de mecanismos paracrinos en la proliferación de las células sutentaculares por medio de activina A, teniendo como resultado la expansión del cordón testicular. Las interacciones entre las distintas poblaciones celulares a través de morfógenos inducen una transición epitelio-mesénquima fundamental en la formación y diferenciación de la gónada masculina.


SUMMARY: Spermatogenesis is a continuous process which starts during the embryo-fetal development. Auto, para and juxtacrine relations indicate the interdependence of the interstitial cells (Leydig) with the peritubular cells (lamina propria) and sustentacular cells (Sertoli). Certain morphogens are fundamental in this process. Sustentacular cells are able to regulate differentiation and function and peritubular interstitial cells through production of IGF1, TGFA, TGFB and DHH. Peritubular cells are able to produce P-Mod-S regulating differentiation sustentacular cells and through FGF2 and FGF9 modulate epithelial-mesenchymal transitions between sustentacular cells and mesonephros. They also remodel the basal membrane of the testicular condom and regulate the differentiation and function of the interstitial cells by means of IGF1, TGFA and TGFB. Interstitial cells are responsible for the production of testosterone and INSL3, influencing male sexual differentiation. It is suggested that they come from mesenchymal cells of the coelomic epithelium and mesonephros. However, other authors propose their origin from cells of the neural crest. These influence through paracrine mechanisms proliferation sutentaculares cells by activin A, resulting in the expansion of cord testicular. The interactions between the different cell populations through morphogens induce a fundamental epithelial-mesenchymal transition in the formation and differentiation of the male gonad.


Subject(s)
Animals , Male , Mice , Connective Tissue Cells/cytology , Sertoli Cells/cytology , Testis/cytology , Testis/embryology , Fetus , Testis/growth & development
14.
An. acad. bras. ciênc ; 89(3): 1745-1751, July-Sept. 2017. tab, graf
Article in English | LILACS | ID: biblio-886766

ABSTRACT

ABSTRACT The germinative, Sertoli and Leydig cells of two caviomorph rodents (Cavia porcellus and Dasyprocta agouti) were counted as well as the estimation of the total volume of the testis and the total volume of seminiferous tubules and interstitium in prepubertal, pubertal and adult animals. The number of spermatogonia, spermatocytes and spermatids cells increased during the pubertal phase in both rodents, notably the spermatid cells. The spermatocyte and spermatid slightly decreased in the adult of both rodents, but the increment in spermatogonia cells number was seen, mainly in cutias. The number of Sertoli cells increased in pubertal rodents, but in the adult the number reduced. Substantial number of Leydig cells was counted in pubertal and adult guinea pigs. In cutias, the number of Leydig cells increased in pubertal phase and decline in adults. The design-based stereological method has proven to be unbiased and reliable to be applied in reproduction studies.


Subject(s)
Animals , Male , Sertoli Cells/cytology , Spermatozoa/cytology , Dasyproctidae/growth & development , Leydig Cells/cytology , Spermatozoa/growth & development , Cell Count , Guinea Pigs
15.
Pesqui. vet. bras ; 37(6): 650-656, jun. 2017. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-895460

ABSTRACT

Flaxseed is considered a functional food with several health benefits. However, because of its high phytoestrogen content, flaxseed influences hormone metabolism and affects the gonadal biomorphology. In this study, computerized histomorphometry was used to evaluate seminiferous and epididymal tubules, considering the different regions of the epididymis (head, body and tail) of rats subjected to a prolonged diet of flaxseed. Young adult male Wistar rats (n=20) were divided into 2 groups during their lactation period: Control Group (CG), fed casein-based meals and Flaxseed Group (FG), fed a 25% flaxseed meal. After 250 days of continuous ingestion, the animals were euthanized and a blood sample was collected. The testicles and epididymis were removed and fixed in buffered formalin solution. The samples were subjected to routine histological paraffin techniques and stained with hematoxilin and eosin. Immunostaining was performed using an antivimentin antibody for Sertoli cell identification. For morphometry, images of the slides were scanned and analyzed using Image J to determine the epithelial height, tubular and luminal diameter and tubular and luminal area. In the hormonal evaluation, FG had a higher serum concentration of estrogen (P=0.001), but no change was observed in the concentration of testosterone. The morphometric assay of seminiferous tubules and epididymal regions revealed no significant differences between the analyzed groups. Similarly, Sertoli cell quantification showed no significant differences in the FG (P=0.98). These results revealed that the continuous and prolonged intake of 25% flaxseed meals from gestation to 250 days of age, even with a significant increase in serum levels of estradiol, does not exert adverse effects on the testicular and epididymal structure or on the cells participating in the spermatogenesis of rats.(AU)


A semente de linhaça é considerada um alimento funcional com vários efeitos benéficos à saúde. Entretanto, devido ao seu elevado teor de fitoestrógenos, esta semente pode influenciar no metabolismo hormonal e interferir na biomorfologia gonadal. Neste estudo, utilizamos a histomorfometria computadorizada para avaliar os túbulos seminíferos e epididimários, considerando as diferentes regiões do epidídimo (cabeça, corpo e cauda) de ratos submetidos a uma dieta prolongada de semente de linhaça. Foram utilizados ratos Wistar machos adultos jovens (n=20) divididos em 2 grupos, durante o período de lactação: Grupo Controle (GC) a base de caseína e Grupo Linhaça (GL) alimentados com 25% de semente de linhaça. Ao final de 250 dias de ingestão contínua, os animais foram sacrificados e amostra de sangue foi coletada. Os testículos e epidídimos foram retirados e fixados em formol tamponado. As amostras foram submetidas ao processamento histológico de rotina para parafina e coradas em hematoxilina e eosina. Foi feita a imunomarcação com anticorpo antivimentina para identificação das células de Sertoli. Para morfometria, as imagens das lâminas foram digitalizadas e analisadas pelo software ImageJ para obtenção dos dados de altura epitelial, diâmetro e área tubular e luminal. Na avaliação hormonal o GL teve maior concentração de estrógeno sérico (p=0,001), mas nenhuma mudança na concentração de testosterona foi observada. Nos parâmetros morfométricos dos túbulos seminíferos e das regiões epididimárias, não houve diferenças significativas entre os grupos analisados. Da mesma forma, a quantificação das células de Sertoli não apresentaram diferenças significativas no GL (p=0,98). Estes resultados mostraram que o consumo contínuo e prolongado de 25% de semente de linhaça desde período gestacional até 250 dias de idade, mesmo com o aumento significativo nos níveis séricos de estradiol, não exerceram efeitos adversos sobre a estrutura testicular e epididimária, assim como nas células participantes da espermatogênese em ratos.(AU)


Subject(s)
Animals , Male , Rats , Seeds , Testis/anatomy & histology , Rats, Wistar/anatomy & histology , Flax/adverse effects , Epididymis/anatomy & histology , Seminiferous Tubules/anatomy & histology , Sertoli Cells , Vimentin , Histological Techniques/veterinary , Phytoestrogens/adverse effects
16.
National Journal of Andrology ; (12): 963-968, 2017.
Article in Chinese | WPRIM | ID: wpr-812849

ABSTRACT

Objective@#To investigate the expression characteristics of the USP24 gene in the mouse testis and its role in spermatogenesis.@*METHODS@#We examined the expression characteristics of USP24 in the testis tissues of wild-type mice at different postnatal weeks (PNW) and androgen receptor (AR)-knockout (ARKO) adult mice using real-time quantitative PCR and immunofluorescence, and detected the transcriptional activity of the USP24 promoter by dual-luciferase reporter gene assay.@*RESULTS@#The expression of the USP24 gene was low in the testis tissue of the wild-type mice at PNW 1, increased dramatically at PNW 3 and stayed at a similar level till PNW 8. The USP24 protein was located mainly in the cytoplasm of Sertoli and spermatogenic cells. Compared with the wild-type, the adult ARKO mice showed a decreased expression of USP24 localized in the posterior head and mid-piece of the mature sperm in the testis. Dual-luciferase reporter gene assay showed that the transcriptional activity of the USP24 promoter was increased after testosterone stimulation.@*CONCLUSIONS@#The increased expression of the USP24 gene was associated with the initiation of sexual development, and the USP24 protein was expressed in the mature sperm of the mice. USP24 is an AR-target gene, which may be involved in the regulation of spermatogenesis in mice.


Subject(s)
Animals , Male , Mice , Mice, Knockout , Promoter Regions, Genetic , Receptors, Androgen , Genetics , Sertoli Cells , Spermatogenesis , Genetics , Spermatozoa , Metabolism , Testis , Metabolism , Testosterone , Transcription, Genetic , Ubiquitin Thiolesterase , Genetics , Metabolism
17.
National Journal of Andrology ; (12): 103-109, 2017.
Article in Chinese | WPRIM | ID: wpr-812802

ABSTRACT

Objective@#To investigate the expression characteristic of the Daxx gene in the mouse testis and its role in spermatogenesis.@*METHODS@#Real-time PCR, Western blot and immunofluorescence were used in examining the expression characteristics of DAXX in the testis tissue from wild-type, Sertoli cell-specific androgen receptor knockout (SCARKO) and androgen receptor knockout (ARKO) mice at different postnatal weeks .@*RESULTS@#The Daxx gene was highly expressed in the testis tissue and mainly in the nuclei of the wild-type mice at 4 postnatal weeks. Compared with the wild-type, the ARKO mice showed a markedly decreased expression of DAXX (0.299±0.026), which displayed a polar distribution in the spermatogenic cells (0.853±0.058) and exhibited no significant difference in the SCARKO mice (1.000±0.015).@*CONCLUSIONS@#The Daxx gene expression is the highest in the middle-stage development of the mouse testis, significantly decreased in ARKO mice as compared with the wild-type, and its location influenced by specific AR knockout in Sertoli cells. DAXX may be involved in the regulation of spermatogenesis in mice.


Subject(s)
Animals , Carrier Proteins , Genetics , Metabolism , Cell Nucleus , Genetics , Metabolism , Gene Expression , Intracellular Signaling Peptides and Proteins , Genetics , Metabolism , Male , Mice , Mice, Knockout , Molecular Chaperones , Nuclear Proteins , Genetics , Metabolism , Receptors, Androgen , Genetics , Sertoli Cells , Spermatogenesis , Genetics , Testis , Metabolism
18.
National Journal of Andrology ; (12): 497-502, 2017.
Article in Chinese | WPRIM | ID: wpr-812735

ABSTRACT

Objective@#To explore the mechanisms of oxidative stress-induced damage to TM4 Sertoli cells in the mouse using metabolomics techniques based on gas chromatography-mass spectrometry (GC-MS).@*METHODS@#We established the model of oxidative stress-induced damage to mouse TM4 Sertoli cells by treatment with H₂O₂. Then, we detected the survival rate and apoptosis rate of the TM4 cells by MTT and flow cytometry respectively, measured the concentration of ROS in the TM4 cells with the DCFH-DA fluorescent probe, and determined the levels of endogenous metabolites in the TM4 cells by GC-MS after H₂O₂ intervention.@*RESULTS@#After 2 hours of treatment with H₂O₂ at 600 μmol/L, the survival rate of the TM4 cells was reduced to about 50%, and the total apoptosis rates in the low- (100 μmol/L), medium- (300 μmol/L), and high-dose (600 μmol/L) groups were (19.45 ± 0.53), (20.12 ± 0.58), and (37.13 ± 0.35)%, respectively, increased in a dose-dependent manner as compared with (10.28 ± 0.35)% in the blank control (P 1, P <0.05).@*CONCLUSIONS@#Oxidative stress-induced damage and apoptosis of TM4 Sertoli cells are closely associated with the metabolism of amino acid, glucose, and energy in the cells.


Subject(s)
Amino Acids , Metabolism , Animals , Apoptosis , Cell Survival , Dose-Response Relationship, Drug , Energy Metabolism , Gas Chromatography-Mass Spectrometry , Glucose , Metabolism , Hydrogen Peroxide , Pharmacology , Male , Metabolomics , Mice , Oxidative Stress , Reactive Oxygen Species , Metabolism , Sertoli Cells , Metabolism , Time Factors
19.
National Journal of Andrology ; (12): 510-516, 2017.
Article in Chinese | WPRIM | ID: wpr-812733

ABSTRACT

Objective@#To explore the role of TGF-β1 in the proliferation and apoptosis of Sertoli cells and its effect on the expressions of tight junction-related proteins and genes in rats.@*METHODS@#Rat Sertoli cells were isolated in vitro, primarily cultured, and divided into groups A (blank control), B (TGF-β1 receptor blocker), C (TGF-β1), and D (TGF-β1 + receptor blocker). The proliferation and apoptosis of the cells were detected by CCK-8 and flow cytometry, respectively. After establishment of the dual-chamber model for the primary culture of Sertoli cells, the trans-epithelia electrical resistance (TER) value was measured and the relative expressions of Occludin, ZO-1 and Claudin Ⅱ determined by RT-PCR and Western blot.@*RESULTS@#The OD value of the proliferation of the Sertoli cells was markedly higher in group C than in groups A and D (0.79 ± 0.04 vs 0.66 ± 0.05 and 0.68 ± 0.02, P0.05). The TER value was dramatically decreased in group C as compared with groups A and D ([176.37 ± 16.61] vs [281.42 ± 9.83] and [254.37 ± 13.55] /cm2, P0.05) or their protein expressions (F = 0.28 and 1.31, P>0.05). Both the mRNA and protein expressions of Occludin were markedly lower in group C than in A and D (P<0.01 and P<0.05), with statistically significant differences among the four groups (F = 6.86 and 6.87, P<0.01).@*CONCLUSIONS@#TGF-β1 can promote the proliferation of Sertoli cells in rats and act on the tight junction of the cells by regulating the expression of Occludin.


Subject(s)
Animals , Apoptosis , Cell Proliferation , Cells, Cultured , Claudin-2 , Metabolism , Male , Occludin , Metabolism , RNA, Messenger , Rats , Sertoli Cells , Cell Biology , Physiology , Tight Junction Proteins , Metabolism , Tight Junctions , Genetics , Metabolism , Transforming Growth Factor beta1 , Physiology , Zonula Occludens-1 Protein , Metabolism
20.
National Journal of Andrology ; (12): 531-535, 2017.
Article in Chinese | WPRIM | ID: wpr-812729

ABSTRACT

Objective@#To investigate the relationship between the serum anti-Müllerian hormone (AMH) level and semen parameters.@*METHODS@#We collected the data about 726 outpatients at the Male Infertility Clinic of Jinling Hospital from September 2015 to November 2016, including 72 with non-obstructive azoospermia, 123 with oligospermia, and 531 with normal sperm concentration. We obtained the semen volume, total sperm count, sperm concentration, sperm motility, the percentages of progressively motile sperm (PMS) and morphologically normal sperm (MNS), and the levels of serum AMH, inhibin B (INH-B), total testosterone (TT) and follicle - stimulating hormone (FSH) of the patients, analyzed the correlation of the serum AMH level with the other parameters, and compared the AMH level among different groups.@*RESULTS@#The serum AMH level was found to be correlated positively with the total sperm count (r = 0.227, P 0.05). There was a statistically significant difference in the serum AMH level among the patients with non-obstructive azoospermia, oligozoospermia, and normal sperm concentration ([6.33 ± 4.26] vs [8.26 ± 3.98] vs [9.8 ± 5.19] ng/ml, P <0.001).@*CONCLUSIONS@#Serum AMH is a biomarker reflecting the function of Sertoli cells and its level is significantly correlated with sperm concentration and motility, suggesting that AMH may be involved in spermatogenesis and sperm maturation.


Subject(s)
Anti-Mullerian Hormone , Blood , Azoospermia , Blood , Biomarkers , Blood , Follicle Stimulating Hormone , Blood , Humans , Inhibins , Blood , Male , Oligospermia , Blood , Semen , Semen Analysis , Sertoli Cells , Physiology , Sperm Count , Sperm Motility , Spermatogenesis , Spermatozoa , Testosterone , Blood
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