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Lactobacillus rhamnosus is a type of bacteria known as a probiotic and is often used to support the health of the digestive system and vaginal flora. This type of bacteria has an important role, showing positive effects on female reproductive biology, particularly by maintaining the balance of microorganisms in the vagina, reducing the risk of infection, and strengthening the immune system to support maternal health during pregnancy. There are also studies showing that these probiotics prevent maternal obesity and gestational diabetes. Consuming probiotics containing Lactobacillus rhamnosus strains may support the intestinal health of breastfeeding mothers, but they may also contribute to the health of offspring. Therefore, this review focuses on the current available data for examining the effects of Lactobacillus rhamnosus strains on female reproductive biology and offspring health. A systematic search was conducted in the PubMed and Web of Science databases from inception to May 2024. The search strategy was performed using keywords and MeSH (Medical Subject Headings) terms. Inconsistent ratings were resolved through discussion. This review is strengthened by multiple aspects of the methodological approach. The systematic search strategy, conducted by two independent reviewers, enabled the identification and evaluation of all relevant literature. Although there is a limited number of studies with high heterogeneity, current literature highlights the important contribution of Lactobacillus rhamnosus probiotics in enhancing female reproductive health and fertility. Furthermore, the probiotic bacteria in breast milk may also support the intestinal health of newborn, strengthen the immune system, and protect them against diseases at later ages.
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PURPOSE: Physiological decidual senescence promotes embryo implantation, whereas pathological decidual senescence causes many pregnancy pathologies. The aim of this study was to evaluate the effect of rapamycin on decidual cell subpopulations and endometrial function in physiological and induced senescence and to investigate the decidual cell subpopulations present in physiological conditions during early pregnancy and implantation in mice. METHODS: Control, physiological decidualization (0.5 mM cAMP and 1 µM MPA added), and induced senescence (0.1 mM HU added) models with and without 200 nM rapamycin treatment were established using a human endometrial stromal cell line, and decidual cell subpopulations were analyzed by immunofluorescence and flow cytometry. The human extravillous trophoblast cell line AC-1M88 was also cultured in decidualization models, and spheroid expansion analysis was performed. In in vivo studies, decidual cell subpopulations were analyzed by immunofluorescence during early mouse pregnancy. RESULTS: The results revealed that rapamycin decreased DIO2 and ß-GAL expressions in physiological and induced senescence without FOXO1. Notably, in induced senescence, increased fragmentation was observed in AC-1M88 cells, and rapamycin treatment successfully attenuated the fragmentation of spheroids. We showed that the FOXO1-DIO2 signaling axis can trigger decidual senescence during early gestation and days of implantation in mice. CONCLUSIONS: Our study underlines the importance of rapamycin in modulating decidual cell subpopulations and endometrial tissue function during decidual senescence. The information obtained may provide insight into the pathologies of pregnancy seen due to decidual senescence and guide better treatment strategies for reproductive problems.
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Consuming alcohol affects almost all organs. Acetaldehyde, formed as the main product as a result of alcohol metabolism, causes the production of free superoxide radicals when oxidized, and accordingly oxidative and apoptotic processes are triggered. There are studies showing that carnitine has effects on oxidative and apoptotic processes that occur in various conditions. However, the mechanisms showing the effects of L-carnitine on these effects of alcohol have not been fully elucidated. In our study, the effects of acetyl-L-carnitine administration on the molecular mechanisms of oxidative stress, endoplasmic reticulum stress, and apoptotic parameters in gastric tissue of rats chronically exposed to alcohol were investigated. Hematoxylin-eosin staining was used for histopathological studies. Endoplasmic reticulum stress markers were detected with immunohistochemical staining and western blotting. Apoptotic index was evaluated by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Total oxidant and antioxidant status were examined by ELISA. Our results showed that chronic alcohol administration caused a significant increase in TOS levels, an indicator of oxidative stress, the levels of ER-stress-associated proteins XBP1, GRP78, and CHOP, and % apoptotic index values in rat gastric tissues. Additionally, it was determined that acetyl-L-carnitine administration caused an improvement in those values. Based on our data, we can conclude that acetyl-L-carnitine has a tissue protective effect by scavenging free oxygen radicals and reducing ER stress-related proteins XBP1, GRP78, and CHOP and apoptosis in chronic ethanol-administered rats, and that this natural antioxidant may be beneficial in the treatment of oxidative stress-induced diseases.
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Acetilcarnitina , Chaperón BiP del Retículo Endoplásmico , Ratas , Animales , Acetilcarnitina/farmacología , Etanol/toxicidad , Antioxidantes/farmacología , Antioxidantes/metabolismo , Apoptosis , Estrés Oxidativo , Proteínas de Choque Térmico/metabolismo , Proteínas de Choque Térmico/farmacología , CarnitinaRESUMEN
PURPOSE: The study aims to investigate first the presence of Syncytin 2 and its receptor, MFSD2, in human sperm, and second whether the expressions of Syncytin 1, Syncytin 2, and their receptors, SLC1A5 and MFSD2, differ between normozoospermic, asthenozoospermic, oligozoospermic, and oligoasthenozoospermic human sperm samples. METHODS: The localization patterns and expression levels of syncytins and their receptors were evaluated in normozoospermic (concentration = 88.9 ± 5.5 × 106, motility = 79.2 ± 3.15%, n = 30), asthenozoospermic (concentration = 51.7 ± 7.18 × 106, motility = 24.0 ± 3.12%, n = 15), mild oligozoospermic (concentration = 13.5 ± 2.17 × 106, motility = 72.1 ± 6.5%, n = 15), moderate oligozoospermic (concentration = 8.4 ± 3.21 × 106, motility = 65.1 ± 8.9%, n = 15), severe oligozoospermic (concentration = 2.1 ± 1.01 × 106, motility = 67.5 ± 3.2%, n = 15), and oligoasthenozoospermic (concentration = 5.5 ± 3.21 × 106, motility = 18.5 ± 1.2%, n = 15) samples by immunofluorescence staining and western blot. RESULTS: Syncytins and their receptors visualized by immunofluorescence showed similar staining patterns with slight staining of the tail in all spermatozoa regardless of normozoospermia, asthenozoospermia, oligozoospermia, or oligoasthenozoospermia. The localization patterns were categorized as equatorial segment, midpiece region, acrosome, and post-acrosomal areas. The combined staining patterns were also detected as acrosomal cap plus post acrosomal region, the midpiece plus equatorial segment, and midpiece plus acrosomal region. However, some sperm cells were categorized as non-stained. Both syncytin proteins were most intensely localized in the midpiece region, while their receptors were predominantly present in the midpiece plus acrosomal region. Conspicuously, syncytins and their receptors showed decreased expression in asthenozospermic, oligozoospermic, and oligoasthenozoospermic samples compared to normozoospermic samples. CONCLUSION: The expression patterns of HERV-derived syncytins and their receptors were identical regardless of the spermatozoa in men with normozoospermia versus impaired semen quality. Further, asthenozoospermia, oligozoospermia, and oligoasthenozoospermia as male fertility issues are associated with decreased expression of both syncytins and their receptors.
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Astenozoospermia , Retrovirus Endógenos , Oligospermia , Humanos , Masculino , Análisis de Semen , Astenozoospermia/genética , Astenozoospermia/metabolismo , Oligospermia/genética , Oligospermia/metabolismo , Semen/metabolismo , Retrovirus Endógenos/metabolismo , Espermatozoides/metabolismo , Motilidad Espermática/genética , Antígenos de Histocompatibilidad Menor/metabolismo , Sistema de Transporte de Aminoácidos ASC/metabolismoRESUMEN
The use of electronic devices such as mobile phones has had a long stretch of rapid growth all over the world. Therefore, exposure to radio frequency radiation (RFR) has increased enormously. Here, we aimed to assess the balance between cell death and proliferation and also investigate the involvement of the JNK/p38 MAPK signaling pathway in the testis of rats exposed to 900 MHz RFR in acute and chronic periods (2 h/day, 5 days/week) for 1 or 10 weeks, respectively. The expression of proliferating cell nuclear antigen (PCNA), Bcl-xL, cleaved caspase-3, phosphorylated-JNK (p-JNK), and phosphorylated-p38 (p-p38) was analyzed in line with histopathology and TUNEL analysis in rat testis. There were no histopathological differences between sham and RFR groups in the acute and chronic groups. PCNA expression was not altered between groups in both periods. However, alterations for cleaved caspase-3 and Bcl-xL were observed depending on the exposure period. TUNEL analysis showed a significant increase in the RFR group in the acute period, whereas no difference in the chronic groups for the apoptotic index was reported. In addition, both p-p38 and p-JNK protein expressions increased significantly in RFR groups in both periods. Our study indicated that 900 MHz RFR might result in alterations during acute period exposure for several parameters, but this can be ameliorated in the chronic period in rat testis. Here, we also report the involvement of the p38/JNK-mediated MAPK pathway after exposure to 900 MHz RFR. Hence, this information might shed light in future studies toward detailed molecular mechanisms in male reproduction and infertility.
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Ondas de Radio , Testículo , Animales , Apoptosis , Caspasa 3 , Masculino , Antígeno Nuclear de Célula en Proliferación , Ondas de Radio/efectos adversos , Ratas , Proteínas Quinasas p38 Activadas por MitógenosRESUMEN
CTCFL is expressed in testis, oocytes and embryonic stem cells, and is aberrantly expressed in malignant cells, and is classified as a cancer-testis gene. We have previously shown by using a tetracycline-inducible Ctcfl transgene that inappropriate expression of Ctcfl negatively impacts fetal development and causes early postnatal lethality in the mouse. The affected pups displayed severe vascular abnormalities and localized hemorrhages in the brain evocative of cerebral cavernous malformations (CCM) and arteriovenous malformations (AVM) in humans. Thus, we aim to analyze; a) the presence of CCM-related proteins CCM1/KRIT1, CCM2/malcavernin and CCM3/PDCD10 in Ctcfl transgenic animals and, b) whether there is CTCFL expression in human CCM and AVM tissues. Ctcfl transgenic animals exhibited increased CD31 expression in vascular areas of the dermis and periadnexal regions but no difference was observed for vWF and α-SMA expressions. CCM-related proteins CCM1/KRIT1, CCM2/malcavernin and CCM3/PDCD10 were aberrantly expressed in coronal sections of the head in transgenic animals. We also observed CTCFL expression in human CCMs and AVMs. The induced expression of CTCFL resulting in vascular brain malformations in mice combined with the presence of CTCFL in human vascular malformations provide new insights into the role of this gene in vascular development in humans.
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Proteínas de Unión al ADN/metabolismo , Hemangioma Cavernoso del Sistema Nervioso Central/metabolismo , Animales , Antígenos CD34/metabolismo , Malformaciones Arteriovenosas/metabolismo , Malformaciones Arteriovenosas/patología , Vasos Sanguíneos/patología , Proteínas de Unión al ADN/genética , Genotipo , Hemangioma Cavernoso del Sistema Nervioso Central/patología , Ratones Transgénicos , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Transgenes , Factor de von Willebrand/metabolismoRESUMEN
The circadian system regulates the daily temporal organization in behavior and physiology, including neuroendocrine rhythms and reproduction. Modern life, however, increasingly impacts this complex biological system. Due to limitations of working with human subjects exposed to shift work schedules, most chronoregulation research has used rodent models. Recent publications in these model systems have emphasized the negative effects of circadian rhythm disruption on both female and male reproductive systems and fertility. Additionally, there is growing concern about the long-term effects of circadian rhythm disruptions during pregnancy on human offspring and their descendants as circadian regulation during pregnancy can also alter epigenetic programing in offspring. However, to truly know if such concerns apply to humans will require retrospective and prospective human studies. Therefore, this review will highlight the latest available evidence regarding potential effects of chronodisruption on both female and male reproductive systems. Additionally, it presents a comprehensive summary of transgenerational and epigenetic effects on adult offspring that result from maternal chronodisruption.
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Ritmo Circadiano , Epigenoma , Feto/fisiopatología , Efectos Tardíos de la Exposición Prenatal/fisiopatología , Reproducción , Animales , Femenino , Humanos , Embarazo , Efectos Tardíos de la Exposición Prenatal/genéticaRESUMEN
Recent developments in the field of insecticide exposure have led to a renewed interest in alternative antioxidant therapy. The present study was to investigate the neuroprotective role of syringic acid (SA, 25â¯mg/kg/day) on the neurotoxicity and oxidative damage induced by deltamethrin (DTM, 1.28â¯mg/kg/day during two months) in CA1/3 pyramidal neurons. Animals were divided into 4 groups (nâ¯=â¯16/group) (250-270â¯g) for control, DTM, SA and DTMâ¯+â¯SA. DTM and SA were administered by oral gavage daily. Rats that were given sub-chronic DTM had revealed a significant increase in caspase-3 levels, impaired recognition memory, reduced antioxidant activity and enhanced free radicals in the hippocampus. The results showed that SA ameliorated neurobehavioral alterations, reduced reactive oxygen/nitrogen species, pyknosis in the CA1/3 and increased antioxidant enzyme activity. In conclusion, SA (25â¯mg/kg/day) had potential neuroprotective and therapeutic impacts against sub-chronic DTM exposure via its antioxidant and antiapoptotic efficacy. Therefore, it can be used as a neuroprotective natural plant-derived agent against DTM-induced neurotoxicity.
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Ácido Gálico/análogos & derivados , Hipocampo/patología , Insecticidas/toxicidad , Trastornos Mentales/prevención & control , Fármacos Neuroprotectores/uso terapéutico , Síndromes de Neurotoxicidad/prevención & control , Nitrilos/toxicidad , Piretrinas/toxicidad , Animales , Apoptosis/efectos de los fármacos , Región CA1 Hipocampal/metabolismo , Región CA1 Hipocampal/patología , Región CA3 Hipocampal/metabolismo , Región CA3 Hipocampal/patología , Ácido Gálico/uso terapéutico , Hipocampo/metabolismo , Aprendizaje/efectos de los fármacos , Masculino , Memoria/efectos de los fármacos , Trastornos Mentales/inducido químicamente , Trastornos Mentales/metabolismo , Síndromes de Neurotoxicidad/psicología , Estrés Oxidativo/efectos de los fármacos , Células Piramidales , Ratas , Ratas WistarRESUMEN
PURPOSE: The purpose of this study is to investigate whether erythrocyte-sperm separation medium (ESSM) has effects on human sperm motility, morphology, viability, membrane maturity, acrosome integrity, and nuclear attributes before and after cryopreservation. METHODS: Semen samples from normozoospermic (n = 36) and oligozoospermic (n = 9) patients were analyzed. Samples from the same patient were divided into three aliquots: group 1 and group 2 were resuspended in sperm washing media and ESSM, respectively. Group 3 was resuspended in ESSM with blood sample to mimic the extensive number of erythrocytes in the testicular sperm extraction (TESE) material. All groups were evaluated for sperm concentration, motility, Kruger/Tygerberg strict morphology, viability by eosin-nigrosin staining, membrane maturity by hyaluronic acid-binding assay (HBA), acrosomal integrity by Pisum sativum lectin staining, chromatin maturity by aniline blue staining, and DNA integrity by TUNEL assay before and after cryopreservation. RESULTS: No significant difference was determined between ESSM-treated and ESSM-untreated sperm samples for the sperm parameters tested (p > 0.05). After cryopreservation, total sperm motility and viability decreased regardless of ESSM used. The percentages of sperm with Tygerberg normal morphology, intact acrosome, and HA-bound sperm were found to be lower in oligozoospermic samples before cryopreservation in each group. However, no statistically significant differences were found between oligozoospermic and normozoospermic samples when all groups were compared. Thus, ESSM treatment did not cause a significant change on sperm motility, normal morphology, viability, HA-binding capacity, chromatin maturity, and DNA fragmentation. CONCLUSION: ESSM can enhance the efficiency of sperm retrieval protocol and can also decrease the time required to collect spermatozoa while not affecting sperm morphogenetic properties.
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Separación Celular/métodos , Eritrocitos , Recuperación de la Esperma , Espermatozoides/citología , Acrosoma , Adolescente , Adulto , Membrana Celular , Cromatina , Criopreservación , Fragmentación del ADN , Humanos , Ácido Hialurónico/metabolismo , Masculino , Persona de Mediana Edad , Oligospermia/patología , Preservación de Semen , Motilidad Espermática , Espermatozoides/fisiologíaRESUMEN
PURPOSE: Our aim was the development of a safe sperm cryopreservation New Media (NM), composed of consistent and reproducible components devoid of any animal origin, and evaluation of NM in terms of its effect on sperm structure and function as compared to regularly used yolk media (TYM) (Irvine Scientific). METHODS: We evaluated patient semen samples and cryopreserved them in duplicates in either NM or TYM. The samples were cryopreserved for either a short term of 1 week or long term of 1 month prior to thawing. The parameters investigated include sperm motility via computer-assisted semen analysis (CASA), sperm concentration, and sperm biomarkers that promote paternal contribution of spermatozoa to fertilization including hyaluronic acid binding, chromatin maturity, apoptotic markers, cytoplasmic retention, and sperm DNA integrity. RESULTS: As compared to TYM, NM was equally capable of sperm cryopreservation with both short-term and long-term storage in media, and after freeze-thaw and gradient processing of sperm. HA binding of sperm was comparable post thaw in both NM and yolk media. There are also no differences observed between the samples cryopreserved in NM or TYM in terms of their aniline blue staining, CK immunocytochemistry, caspase 3 immunostaining, or DNA nick translation. CONCLUSIONS: NM has the advantage of being xeno-free, yet in preservation of sperm motility and other sperm attributes, the NM is as effective as the TYM.
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Criopreservación , Análisis de Semen , Preservación de Semen/métodos , Espermatozoides/crecimiento & desarrollo , Animales , Cromatina/efectos de los fármacos , Crioprotectores/farmacología , Congelación , Humanos , Masculino , Recuento de Espermatozoides , Motilidad Espermática/efectos de los fármacos , Espermatozoides/efectos de los fármacosRESUMEN
Human life begins with sperm and oocyte fusion. After fertilization, various fusion events occur during human embryogenesis and morphogenesis. For example, the fusion of trophoblastic cells constitutes a key process for normal placental development. Fusion in the placenta is facilitated by syncytin 1 and syncytin 2. These syncytins arose from retroviral sequences that entered the primate genome 25 million and more than 40 million years ago respectively. About 8% of the human genome consists of similar human endogenous retroviral (HERVs) sequences. Many are inactive because of mutations or deletions. However, the role of the few that remain transcriptionally active has not been fully elucidated. Syncytin proteins maintain cell-cell fusogenic activity based on ENV: gene-mediated viral cell entry. In this review, we summarize how syncytins and their receptors are involved in fusion events during human reproduction. The significance of syncytins in tumorigenesis is also discussed.
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Productos del Gen env/metabolismo , Neoplasias/patología , Placentación , Proteínas Gestacionales/metabolismo , Reproducción/fisiología , Animales , Femenino , Humanos , Neoplasias/metabolismo , Embarazo , RetroviridaeRESUMEN
INTRODUCTION: Syncytins belong to the Human Endogenous Retrovirus family. The syncytin-1 receptor, SLC1A5, and syncytin-2 receptor, MFSD2, interact with their respective syncytin proteins to induce syncytiotrophoblast formation. However, there is no information about syncytins in gestational diabetic placenta. Therefore, we studied the expression and localization of syncytins and their receptors during normal placental development and in gestational diabetic placenta. METHODS: Immunohistochemistry and Western-blot methods were performed with antibodies against syncytin-1, syncytin-2, SLC1A5 and MFSD2 in human first trimester placental tissues, normal term and gestational diabetic placentas. Syncytin-1, syncytin-2 and MFSD2 mRNA transcripts were determined by qRT-PCR in normal and diabetic term placentas. RESULTS: Cytoplasmic syncytin-1, syncytin-2, SLC1A5 and MFSD2 immunoreactions were observed in the trophoblastic layers in all placental samples. Some of the stromal cells showed strong cytoplasmic punctate staining. There were significantly weak syncytin-2 and MFSD2 immunoreaction intensities in diabetic placentas by ImageJ analysis, in parallel with decreased syncytin-2 and MFSD2 proteins in diabetic placentas by Western-blot. Protein expression of SLC1A5 increased dramatically in early pregnancy compared to term placenta. Syncytin-1, syncytin-2 and MFSD2 mRNA transcripts showed similar relative expression pattern by qRT-PCR. DISCUSSION: Syncytins were localized not only in cytotrophoblast cells and the basement membrane of the syncytiotrophoblast but also in the apical microvillous membrane and cytoplasm of syncytiotrophoblast, and some of the stromal cells and endothelium. Decreased syncytin-2 and MFSD2 proteins in gestational diabetic placentas might cause abnormal syncytiotrophoblast formation and possibly be involved in the pathology. Therefore, our study highlights an important potential relationship between syncytins and gestational diabetic placenta.
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Sistema de Transporte de Aminoácidos ASC/biosíntesis , Productos del Gen env/biosíntesis , Antígenos de Histocompatibilidad Menor/biosíntesis , Placenta/metabolismo , Proteínas Gestacionales/biosíntesis , Embarazo en Diabéticas/patología , Adulto , Western Blotting , Retrovirus Endógenos , Femenino , Humanos , Inmunohistoquímica , Embarazo , Embarazo en Diabéticas/metabolismo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Mammalian target of rapamycin (mTOR) signaling serves as a central regulator of cell growth, proliferation, and survival by interacting with various proteins. To date, few studies implicated mTOR in placenta. Human placenta in gestational diabetes mellitus (GDM) shows several alterations including villous immaturity, impaired placental function, and overgrowth. Hence, we aimed to investigate the expression of mTOR, phospho-mTOR (p-mTOR), and the 2 phosphorylated downstream targets of mTOR, ribosomal protein S6 kinase 1 (p-p70S6K), and eukaryotic initiation factor 4E-binding protein 1 (p-4EBP1) in normal term and gestational diabetic human placentas. Immunohistochemistry and Western blot were performed with antibodies against mTOR, p-mTOR, p-p70S6K, and p-4EBP1 (Thr37/46) in normal and diabetic placentas (n = 6 each) and quantified by ImageJ. All mTOR pathway components that we studied were immunolocalized in both normal and diabetic placenta groups. Syncytiotrophoblast and the vascular wall in villi displayed cytoplasmic mTOR and p-mTOR (S2448) immunoreactivities in all placenta samples. However, increased expression of p70S6K in syncytiotrophoblast and p-4EBP1 (Thr37/46) in villous stromal cells was observed in gestational diabetic placentas. Western blot analysis also confirmed the statistically significant increase in p-p70S6K (T389) expression in diabetic placentas. The altered expression of downstream components of mTOR signaling in gestational diabetic placentas suggests an involvement of mTOR activity in the placental pathology of GDM. However, whether increased nutrient transport via this pathway will stimulate fetal and placental overgrowth is still unknown. Although this is a descriptive study, further studies with a functional analysis to highlight the molecular mechanisms underlying this placental pathology are proposed.
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Diabetes Gestacional/metabolismo , Regulación del Desarrollo de la Expresión Génica , Placenta/metabolismo , Serina-Treonina Quinasas TOR/biosíntesis , Adulto , Diabetes Gestacional/patología , Femenino , Humanos , Placenta/patología , Embarazo , Serina-Treonina Quinasas TOR/genéticaRESUMEN
CTCFL, a paralog of CTCF, also known as BORIS (brother of regulator of imprinted sites), is a testis-expressed gene whose function is largely unknown. Its product is a cancer testis antigen (CTA), and it is often expressed in tumor cells and also seen in two benign human vascular malformations, juvenile angiofibromas and infantile hemangiomas. To understand the function of Ctcfl, we created tetracycline-inducible Ctcfl transgenic mice. We show that Ctcfl expression during embryogenesis results in growth retardation, eye malformations, multiorgan pathologies, vascular defects, and neonatal death. This phenotype resembles prior mouse models that perturb the transforming growth factor ß (TGFB) pathway. Embryonic stem (ES) cells with the Ctcfl transgene reproduce the phenotype in ES cell-tetraploid chimeras. Transcriptome sequencing of the Ctcfl ES cells revealed 14 genes deregulated by Ctcfl expression. Bioinformatic analysis revealed the TGFB pathway as most affected by embryonic Ctcfl expression. Understanding the consequence of Ctcfl expression in nontesticular cells and elucidating downstream targets of Ctcfl could explain the role of its product as a CTA and its involvement in two, if not more, human vascular malformations.
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Proteínas de Unión al ADN/metabolismo , Retardo del Crecimiento Fetal/metabolismo , Factor de Crecimiento Transformador beta/fisiología , Animales , Animales Recién Nacidos , Encéfalo/metabolismo , Encéfalo/patología , Proteínas de Unión al ADN/genética , Femenino , Retardo del Crecimiento Fetal/patología , Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Genes Letales , Masculino , Ratones Endogámicos C57BL , Ratones Transgénicos , Transducción de SeñalRESUMEN
The aim of this study was to assess the effect of oral magnesium supplementation (Mg-supp) on blood pressure (BP) and possible mechanism in nitric oxide synthase (NOS) inhibition-induced hypertension model. Hypertension and/or Mg-supp were created by N-nitro-l-arginine methyl ester (25 mg/kg/day by drinking water) and magnesium-oxide (0.8% by diet) for 6 weeks. Systolic BP was measured weekly by tail-cuff method. The effects of hypertension and/or Mg-supp in thoracic aorta and third branch of mesenteric artery constriction and relaxation responses were evaluated. NOS-inhibition produced a gradually developing hypertension and the magnitude of the BP was significantly attenuated after five weeks of Mg-supp. The increased phenylephrine-induced contractile and decreased acetylcholine (ACh)-induced dilation responses were found in both artery segments of hypertensive groups. Mg-supp was restored ACh-relaxation response in both arterial segments and also Phe-constriction response in thoracic aorta but not in mesenteric arteries. The contributions of NO, prostaglandins and K(+) channels to the dilator response of ACh were similar in the aorta of all the groups. The contribution of the NO to the ACh-mediated relaxation response of mesenteric arteries was suppressed in hypertensive rats, whereas this was corrected by Mg-supp. The flow-mediated dilation response of mesenteric arteries in hypertensive rats failed and could not be corrected by Mg-supp. Whereas, vascular eNOS protein and magnesium levels were not changed and plasma nitrite levels were reduced in hypertensive rats. The results of this study showed that Mg-supp lowered the arterial BP in NOS-inhibition induced hypertension model by restoring the agonist-induced relaxation response of the arteries.
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Presión Sanguínea/fisiología , Hipertensión/fisiopatología , Magnesio/administración & dosificación , Óxido Nítrico Sintasa/antagonistas & inhibidores , Animales , Presión Sanguínea/efectos de los fármacos , Modelos Animales de Enfermedad , Inhibidores Enzimáticos/farmacología , Hipertensión/etiología , Hipertensión/metabolismo , Masculino , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico Sintasa/metabolismo , Ratas , Ratas WistarRESUMEN
PURPOSE: To develop a method for delayed assessment of sperm motility, after shipment of semen to a remote laboratory. Sperm in semen were labeled with the MitoTracker(®) Red CM-H(2)XRos reagent, and fixed with 3.7 % formaldehyde by the laboratory technicians at the origin of the semen. This treatment reflected well sperm mitochondrial activity, and the MitoTracker(®) signal was related to sperm motility and velocity for 2-3 days following ejaculation. METHODS: Sperm motility and velocity were evaluated manually and by computer assisted semen analysis (CASA), respectively. Fluorescence assessment of individual sperm was carried out with the computer assisted Metamorph v4.6.9 program. Emission levels of MitoTracker(®) spermatozoa were studied in room temperature and cooled semen, or in the respective room temperature swim-up sperm fractions following ejaculation, and on the second day (N = 103 samples, 89 men) and third day (N = 10 samples, 8 men). RESULTS: Sperm with optical density (O.D.) ≥0.7 showed close correlations with ejaculatory sperm motility and velocity even after second day (r = 0.92, p < 0.001, N = 103 samples). Further, the multiple of sperm motility and velocity was also related to the proportion of high MitoTracker(®) reagent emission sperm (r = 0.83, p < 0.001, N = 103 samples). MitoTracker(®) dye fluorescence on the second day accurately reflected the ejaculatory sperm motility (r = 0.90, p < 0.001). Thus, a shipping delay would not adversely affect the results. CONCLUSIONS: The delayed assessment of sperm motility in samples treated with MitoTracker(®) Red CM-H(2)XRos reagent and shipped to remote laboratory truly reflects the level of sperm motility at the time of the ejaculation.
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Preservación de Semen/métodos , Semen/fisiología , Motilidad Espermática/fisiología , Criopreservación , Eyaculación/fisiología , Humanos , Masculino , Análisis de Semen , Recuento de Espermatozoides , EspermatozoidesRESUMEN
In mammalian species, acquisition of sperm fertilization competence is dependent on the phenomenon of sperm capacitation. One of the key elements of capacitation is protein tyrosine phosphorylation (TP) in various sperm membrane regions. In previous studies performed, the pattern of TP was examined in human sperm bound to zona pellucida of oocytes. In the present comparative study, TP patterns upon sperm binding to the zona pellucida or hyaluronic acid (HA) were investigated in spermatozoa arising from the same semen samples. Tyrosine phosphorylation, visualized by immunofluorescence, was localized within the acrosomal cap, equatorial head region, neck, and the principal piece. Tyrosine phosphorylation has increased in a time-related manner as capacitation progressed, and the phosphorylation pattern was identical within the principal piece and neck, regardless of the sperm bound to the zona pellucida or HA. Thus, the data demonstrated that the patterns of sperm activation-related TP were similar regardless of the spermatozoa bound to zona pellucida or HA. Further, sperm with incomplete development, as detected by excess cytoplasmic retention, failed to exhibit TP.
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Ácido Hialurónico/metabolismo , Espermatozoides/metabolismo , Tirosina/metabolismo , Zona Pelúcida/metabolismo , Humanos , Masculino , Fosforilación/fisiología , Unión Proteica/fisiología , Motilidad Espermática/fisiologíaRESUMEN
In this chapter, the laboratory methods for detection of sperm biomarkers that are aimed at identifying arrested sperm development are summarized. These probes include sperm staining with aniline blue for persistent histones, representing a break in the histone-transition protein-protamine sequence, immunocytochemistry with cytoplasmic sperm proteins, highlighting cytoplasmic retention during spermiogenesis, DNA nick translation testing for DNA chain fragmentation due to various reasons, for instance low HspA2 chaperone protein levels, and consequential diminished DNA repair. Finally, we briefly provide references on our work on sperm hyaluronan binding, abnormal Tybergerg sperm morphology, and the increased levels of chromosomal aneuploidies in sperm with developmental arrest. A very interesting aspect of the biomarker field is the discovery (Sati et al, Reprod Biomed Online 16:570-579, 2008) that the various nuclear and cytoplasmic defects detected by the biomarkers are related, and may simultaneously occur within the same spermatozoa as evidenced by a combination of biomarkers, such as aniline blue staining (persistent histones) coupled with cytoplasmic retention, DNA fragmentation, Caspase-3, Tygerberg abnormal morphology, and increased levels of chromosomal aneuploidies. We show examples of this >80% overlap in staining patterns within the same spermatozoa.
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Compuestos de Anilina , Núcleo Celular/metabolismo , Cromatina/metabolismo , Citoplasma/metabolismo , Espermatozoides/metabolismo , Coloración y Etiquetado/métodos , Apoptosis , Caspasa 3/metabolismo , Creatina Quinasa/metabolismo , Humanos , Inmunohistoquímica/métodos , Hibridación Fluorescente in Situ/métodos , Masculino , Análisis de Semen/instrumentación , Análisis de Semen/métodosRESUMEN
Despite the known adverse effects of abamectin pesticide, little is known about its action on male fertility. To explore the effects of exposure to abamectin on male fertility and its mechanism, low (1mg/kg/day) and high dose (4 mg/kg/day) abamectin were applied to male rats by oral gavage for 1week and for 6weeks. Weight of testes, serum reproductive hormone levels, sperm dynamics and histopathology of testes were used to evaluate the reproductive efficiency of abamectin-exposed rats. Abamectin level was determined at high concentrations in plasma and testicular tissues of male rats exposed to this pesticide. The testes weights of animals and serum testosterone concentrations did not show any significant changes after abamectin exposure. Abamectin administration was associated with decreased sperm count and motility and increased seminiferous tubule damage. In addition, significant elevations in the 4-hydroxy-2-nonenal (4-HNE)-modified proteins and poly(ADP-ribose) (PAR) expression, as markers for oxidative stress and poly(ADP-ribose) polymerase (PARP) activation, were observed in testes of rats exposed to abamectin. These results showed that abamectin exposure induces testicular damage and affects sperm dynamics. Oxidative stress-mediated PARP activation might be one of the possible mechanism(s) underlying testicular damage induced by abamectin.
Asunto(s)
Fertilidad/efectos de los fármacos , Insecticidas/toxicidad , Ivermectina/análogos & derivados , Espermatogénesis/efectos de los fármacos , Testículo/efectos de los fármacos , Animales , Epidídimo/efectos de los fármacos , Epidídimo/metabolismo , Epidídimo/patología , Hormona Folículo Estimulante/sangre , Insecticidas/sangre , Insecticidas/farmacocinética , Ivermectina/sangre , Ivermectina/farmacocinética , Ivermectina/toxicidad , Masculino , Estrés Oxidativo , Poli(ADP-Ribosa) Polimerasas/metabolismo , Ratas , Ratas Sprague-Dawley , Recuento de Espermatozoides , Motilidad Espermática/efectos de los fármacos , Testículo/metabolismo , Testículo/patologíaRESUMEN
In various tissues, glucocorticoids (GCs) are known to downregulate glucose transport systems; however, their effects on glucose transporters (GLUTs) in the placenta of a diabetic rat are unknown. Glucocorticoid hormone action within the cell is regulated by the glucocorticoid receptor (GR). Thus, this study was designed to investigate the relationship between GR and glucose transporter expression in the placenta of the diabetic rat. Our immunohistochemical results indicated that GR and glucose transporter protein 1 (GLUT 1) are expressed ubiquitously in the trophoblast and endothelial cells of the labyrinthine zone, where maternal fetal transport takes place in the rat placenta. Expression of GR in the junctional zone of the rat placenta was detected in giant cells, and in some spongiotrophoblast cells, but not in the glycogen cells. GLUT 1 was present, especially in glycogen cells during early pregnancy, and in the spongiotrophoblast cells of the junctional zone during late pregnancy. Amounts of GR and GLUT 1 protein were increased towards the end of gestation both in the control and the diabetic placenta. However, at days 17 and 19 of gestation, only the placental GR protein was significantly increased in the streptozotocin-induced diabetic rats compared to control rats. Diabetes led to a significant decrease in placental weight at gestation day 15. In contrast, at gestational days 17 and 21, the weights of the diabetic placenta were significantly increased as compared with the controls. Moreover, diabetes induced fetus intrauterine growth retardation at gestational days 13, 17 and 21. In conclusion, the localization pattern of GR and GLUT 1 proteins in the same cell types led us to believe that there might be a relationship between GR and GLUT 1 expressions at the cellular level. GLUT 1 does not play a pivotal role in diabetic pregnancies. However, placental growth abnormalities during diabetic pregnancy may be related to the amount of GR.