Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 68
Filter
1.
Heliyon ; 5(12): e02932, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31867458

ABSTRACT

ODF1 has been described as an exclusively expressed testicular protein and is located in the outer dense fibers along the sperm tail. ODF1 has been involved in the sperm motility and in the development of the flagellum, but the function of ODF1 is not already clear. Other ODF proteins, such as ODF2 have been characterized in other tissues like the basal body of the kidney primary cilium, but so far only the mRNA of ODF1 has been described in other tissues. These observations let us to hypothesize that the expression of the protein ODF1 could not be limited to the testis. Therefore, in the present work we proposed to evaluate if the ODF1 protein could also be present in tissues other than the testis. Here we demonstrated through western blot, immunofluorescence, and RT-PCR techniques that the protein and mRNA of ODF1 have been identified in the rat kidney. Finally, the presence of ODF1 in kidney has also been confirmed through proteomic analysis using mass spectrometry. The results derived from these different complementary approaches indicate that, to our knowledge and for the first time, ODF1 is demonstrated to be present in an additional organ different to testis. This results raise new questions about potential other functions and locations of the ODF1 protein.

2.
Heliyon ; 5(11): e02868, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31844747

ABSTRACT

A chronic-positive energetic balance has been directly correlated with infertility in men, but the involved mechanisms remain unknown. Herein we investigated weather in a mouse model a chronic feeding with a diet supplemented with chicken fat affects sperm head morphology. To accomplish this, we fed mice for 16 weeks with either control food (low-fat diet, LFD) or control food supplemented with 22% chicken fat (high-fat diet, HFD). At the end of the feeding regimen, we measured: redox and inflammatory changes, cholesterol accumulation in testis and analyzed testicular morphological structure and ultra-structure and liver morphology. We found that the mice fed HFD resembled some features of the human metabolic syndrome, including systemic oxidative stress and inflammation, this group showed an increment in the following parameters; central adiposity (adiposity index: 1.07 ± 0.10 vs 2.26 ± 0.17), dyslipidemia (total cholesterol: 153.3 ± 2.6 vs 175.1 ± 8.08 mg/dL), insulin resistance (indirect Insulin resistance index, TG/HDL-c: 2.94 ± 0.33 vs 3.68 ± 0.15) and fatty liver. Increased cholesterol content measured by filipin was found in the testicles from HFD (fluorescence intensity increase to 50%), as well as an alteration of spermiogenesis. Most remarkably, a disorganized manchette-perinuclear ring complex and an altered morphology of the sperm head were observed in the spermatozoa of HFD-fed mice. These results add new information to our understanding about the mechanisms by which systemic oxidative stress and inflammation may influence sperm-head morphology and indirectly male fertility.

3.
Inflammation ; 42(5): 1886-1900, 2019 Oct.
Article in English | MEDLINE | ID: mdl-31359324

ABSTRACT

Chicken fat and fructose are added into food-processing to reduce costs and enhance acceptability; however, these additives turn food into unhealthy and hypercaloric meals. Herein we have hypothesized that chronic feeding with chicken fat and fructose, together or by separate, can cause pulmonary redox and inflammatory changes. These changes are particularly related to neutrophils and myeloperoxidase, with consequent changes in the organ histophysiology. To test this hypothesis, we fed mice for 16 weeks with either control food (low-fat diet, LFD) or control food supplemented with 22% chicken fat and with or without 10% fructose in the drinking water. At the end of the feeding regimen, we measured redox and inflammatory changes in the lung with particular emphasis on neutrophil accumulation/activation and molecular-histological markers of fibrosis. Our results suggest that a diet supplemented with chicken fat and fructose causes additive effects on pulmonary oxidative stress, inflammation, and a pro-fibrotic status. Neutrophilic inflammation may play a critical role in pulmonary pathology associated with metabolic syndrome.


Subject(s)
Diet, High-Fat/adverse effects , Fibrosis/etiology , Neutrophils/pathology , Pneumonia/etiology , Animals , Inflammation/etiology , Lung/metabolism , Mice , Oxidation-Reduction , Oxidative Stress , Pneumonia/metabolism , Pneumonia/pathology
4.
Reproduction ; 143(3): 281-95, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22228629

ABSTRACT

Serine protease inhibitor Kazal-type (SPINK3)/P12/PSTI-II is a small secretory protein from mouse seminal vesicle which contains a KAZAL domain and shows calcium (Ca(2+))-transport inhibitory (caltrin) activity. This molecule was obtained as a recombinant protein and its effect on capacitated sperm cells was examined. SPINK3 inhibited trypsin activity in vitro while the fusion protein GST-SPINK3 had no effect on this enzyme activity. The inactive GST-SPINK3 significantly reduced the percentage of spermatozoa positively stained for nitric oxide (NO) with the specific probe DAF-FM DA and NO concentration measured by Griess method in capacitated mouse sperm; the same effect was observed when sperm were capacitated under low Ca(2+) concentration, using either intracellular (BAPTA-AM) or extracellular Ca(2+) (EDTA) chelators. The percentage of sperm showing spontaneous and progesterone-induced acrosomal reaction was significantly lower in the presence of GST-SPINK3 compared to untreated capacitated spermatozoa. Interestingly, this decrease was overcome by the exogenous addition of the NO donors, sodium nitroprusside (SNP), and S-nitrosoglutathione (GSNO). Phosphorylation of sperm proteins in tyrosine residues was partially affected by GST-SPINK3, however, only GSNO was able to reverse this effect. Sperm progressive motility was not significantly diminished by GST-SPINK3 or BAPTA-AM but enhanced by the addition of SNP. This is the first report that demonstrates that SPINK3 modulates sperm physiology through a downstream reduction of endogenous NO concentration and independently of SPINK3 trypsin inhibitory activity.


Subject(s)
Glycoproteins/physiology , Nitric Oxide/metabolism , Prostatic Secretory Proteins/physiology , Spermatozoa/physiology , Animals , Cell Survival/drug effects , Down-Regulation/drug effects , Enzyme Inhibitors/metabolism , Enzyme Inhibitors/pharmacology , Glycoproteins/genetics , Glycoproteins/pharmacology , Male , Mice , Mice, Inbred BALB C , Models, Biological , Nitric Oxide/analysis , Osmolar Concentration , Prostatic Secretory Proteins/genetics , Prostatic Secretory Proteins/pharmacology , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/pharmacology , Semen Analysis , Sperm Capacitation/drug effects , Sperm Capacitation/physiology , Spermatozoa/drug effects , Spermatozoa/metabolism , Trypsin/metabolism , Trypsin/pharmacology , Trypsin Inhibitor, Kazal Pancreatic
5.
Theriogenology ; 76(3): 436-47, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21601269

ABSTRACT

Whole seminal plasma (SP) enhances the function and fertility of frozen/thawed ram sperm. The objective of the current study was to investigate whether SP proteins capable of binding to molecules from the sperm plasma membrane were conserved among ram breeds, and whether these proteins were sufficient to overcome cryopreservation-induced reductions in sperm quality. Whole ram SP, obtained from rams of various breeds, improved progressive motility of frozen/thawed sperm at all times evaluated (P < 0.05); however, it did not improve total motility (15 min, P = 0.480; 30 min, P = 0.764; and 45 min, P = 0.795). To identify SP proteins responsible for this effect, a new method was developed to retain SP proteins that bound specifically to the sperm membrane by immobilization of sperm membrane proteins. These proteins specifically bound to the sperm surface, especially the acrosomal region. Lactotransferrin, epididymal secretory protein E1, Synaptosomal-associated protein 29, and RSVP-20 were identified (mass spectrometry) in this fraction. The retained SP proteins fraction repaired ultrastructural damage of frozen/thawed sperm and, with the addition of fructose, significantly improved motility of frozen/thawed sperm. We concluded that SP proteins that bound to the sperm membrane were conserved among ram breeds, and that when added to frozen/thawed semen (along with an energy source), they repaired ram sperm damage and enhanced sperm motility.


Subject(s)
Cell Membrane/ultrastructure , Cryopreservation/veterinary , Semen/metabolism , Seminal Plasma Proteins/physiology , Sheep , Spermatozoa/ultrastructure , Animals , Cell Membrane/metabolism , Chemical Fractionation , Male , Seminal Plasma Proteins/chemistry , Seminal Plasma Proteins/metabolism , Sperm Motility
6.
Gen Comp Endocrinol ; 166(3): 556-64, 2010 May 01.
Article in English | MEDLINE | ID: mdl-20152839

ABSTRACT

The severe environments where Phymaturus lizards inhabit in the Andes highlands and in Patagonia, Argentina, impose restrictions on their reproduction, offering a framework for the development of life history strategies to overcome hard weather conditions. Among them, prolonged female cycles, asynchrony between sexes in receptivity, and sperm storage in males, were described. Asynchrony in the reproductive timing between males and females is a consequence of different energy requirements for gametogenesis, and often imply the existence of cellular mechanisms to enhance fertilization, such as the asynchronic steroid synthesis between testicular compartments, allowing gametogenesis independently of mating. In the present study ultrastructural and hormone assays were combined for the first time in liolaemids. Specifically, morphological features of steroid activity in Leydig and Sertoli cells, and serum testosterone concentrations have been studied in the lizard Phymaturus antofagastensis. Leydig and Sertoli cells presented morphological features characteristic of steroid synthesis during the spermatogenesis, and evident asynchronic steroid production between testicular compartments. Active Sertoli cells and inactive Leydig cells were observed in spring and autumn, while in mid-summer their steroid activity was synchronic in coincidence with maximal abundance of spermatozoa in epididymis. Serum testosterone concentration was at its maximum in mid-summer (126-230 ng ml(-1)), and minimum in late spring (4-24 ng ml(-1)) and early autumn (2-17 ng ml(-1)). In view of these results, P. antofagastensis males show an original approach to adjust their reproductive activity to physiological and environmental constraints at high latitudes and altitudes in the Andean highlands of Argentina.


Subject(s)
Leydig Cells/ultrastructure , Lizards/metabolism , Lizards/physiology , Sertoli Cells/ultrastructure , Spermatogenesis/physiology , Testosterone/blood , Animals , Male , Microscopy, Electron, Transmission
7.
Acta Otorrinolaringol Esp ; 57(5): 247-50, 2006 May.
Article in Spanish | MEDLINE | ID: mdl-16768204

ABSTRACT

Death following tonsillectomy in children is very rare. It is most commonly due to bleeding or aspiration. But, there is another potentially lethal complication following the pediatric tonsillectomy, post-surgical hyponatremia. Acute hyponatremia can lead to catastrophic neurological sequelae. Although this cmplication is not related to the surgical technique, it is potentially life-threatening. The most important factor for hospital acquired hyponatremia is the administration of excessive amounts of hypotonic fluid in situations in wich antidiuretic hormone (ADH) is secreted for non osmotic reasons. We will discuss the etiology and pathophysiology of post-operative hyponatremia. We expect that fatal post-operative hyponatremia can be avoided in pediatric tonsillectomy patients.


Subject(s)
Hyponatremia/etiology , Tonsillectomy/adverse effects , Cause of Death , Child, Preschool , Female , Humans , Hyponatremia/diagnosis , Hyponatremia/therapy
8.
Theriogenology ; 66(5): 1185-93, 2006 Sep 15.
Article in English | MEDLINE | ID: mdl-16647751

ABSTRACT

Extensive work was done regarding the ability of Swim up and Percoll gradient to select functional sperm for in vitro embryo production (IVP) systems. The aim of this work was to compare Swim up and Percoll as methods of sperm selection by ultrastructural, biochemical and functional studies. Frozen-thawed semen from two bulls (Experiments 1 and 2, respectively) were treated using Swim up or Percoll discontinuous gradients. Motility, sperm membrane ultrastructure, sperm proteins, in vitro embryo production (insemination doses, cleavage, embryo yield and quality) and embryo sex ratio were scored and compared. Electron transmission microscopy of outer sperm membranes showed higher (P<0.05) percentage of sperm with lost acrosomes in Percoll treated samples compared to Swim up. A differential protein pattern was also detected. When in vitro embryo production was performed, Percoll gradient produced higher (P<0.05) number of fertilizing doses (7.6 versus 5.9, Bull 1; 13.5 versus 7.8, Bull 2) and higher sperm motility (90% versus 76.6%, Bull 1; 81.7% versus 68.3%, Bull 2) than Swim up. The percentage of cleavage (Day 3) was similar in both treatment groups, whereas embryo production rate (Day 7) was higher (39.4% versus 30.2%, Bull 1; 38% versus 32.4%, Bull 2; P<0.05) when Percoll gradient was used. The percentage of hatched embryos (Day 11) and sex ratio did not differ. Total cell counting and embryo differential staining (inner cell mass and trophoblast cells) of Day 7 embryos showed that Percoll treated sperm produced better quality embryos compared to Swim up. We concluded that Percoll had a better performance selecting sperm and an enhanced capacity for embryo production when compared with the Swim up procedure; this could be attributed to a better acrosome exocytosis, associated to the absence of certain membrane proteins.


Subject(s)
Cattle/physiology , Cell Separation/veterinary , Embryo Culture Techniques/veterinary , Spermatozoa/physiology , Spermatozoa/ultrastructure , Acrosome/physiology , Acrosome/ultrastructure , Animals , Cell Separation/methods , Centrifugation, Density Gradient/veterinary , Cryopreservation/methods , Cryopreservation/veterinary , Embryo Culture Techniques/methods , Female , Fertilization in Vitro/veterinary , Male , Microscopy, Electron, Transmission/methods , Microscopy, Electron, Transmission/veterinary , Microscopy, Fluorescence , Oocytes , Povidone/pharmacology , Semen Preservation/methods , Semen Preservation/veterinary , Sex Ratio , Silicon Dioxide/pharmacology , Sperm Count/veterinary , Sperm Motility
9.
Reproduction ; 129(3): 291-8, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15749956

ABSTRACT

Bovine sperm protease, 66 kDa (BSp66) is a serine protease previously characterized in bovine spermatozoa. Like other proteases, it may be present in sperm from other mammalian species different from bovine, playing a role in the fertilization process. In this study, we looked for BSp66 in hamster spermatozoa using heterologous antibodies against bovine BSp66. An immunoreactive protein was detected by Western blotting in mature and immature sperm. The detected protein had two isoforms similar to the ones reported in bovine sperm. Furthermore, indirect immune detection by fluorescence and electron microscopy assays, showed BSp66 signal at the acrosomal region similar to bovine sperm. As it was determined in bovine sperm, the acrosomal reaction displays the antigen within the acrosomal content. When live hamster sperm was incubated with polyclonal antibody against bovine BSp66 a decrease in the number of sperm bound to zona pellucida in homologous IVF and an impairment of head-head agglutination, were observed. These results suggest that a protease homologous to bovine BSp66 is present in golden hamster spermatozoa, with a conserved molecular mass and cellular location. Moreover, hamster BSp66 is probably involved in zona pellucida recognition.


Subject(s)
Cricetinae/metabolism , Serine Endopeptidases/metabolism , Sperm-Ovum Interactions/physiology , Spermatozoa/metabolism , Animals , Cattle , Cell Culture Techniques , Cell Survival , Fertilization in Vitro , Immunohistochemistry/methods , Male , Microscopy, Immunoelectron , Sperm Motility/physiology
10.
Biochem Biophys Res Commun ; 324(2): 874-7, 2004 Nov 12.
Article in English | MEDLINE | ID: mdl-15474509

ABSTRACT

Fertilization in mammals comprises a sequence of events leading to the fusion of sperm and oocyte membranes. Although proteases are known to be involved in this process, their role in fertilization is controversial. There is extensive work on the characterization of proteolytic systems, including serine proteases, which demonstrates that acrosomal proteases can be distinguished among the sperm of different mammalian species on the basis of the gelatin-hydrolyzing activity on SDS-PAGE by the quantity and variety of the enzymes. In this report, we investigated the occurrence and activity of the serine protease BSp66, previously characterized in bovine spermatozoa, in various mammalian sperm. A protein with a molecular mass of 66 kDa cross-reacted with heterologous antibodies against bovine BSp66 when sperm extracts of several mammalian species were analyzed by Western blot. In agreement, proteolytic activity corresponding to the molecular mass of BSp66 was detected by gelatin zymography in all the species analyzed. This protein was located on the acrosomal region of sperm cells by immunofluorescence methods. We concluded that BSp66 is widespread in mammalian sperm, with a conserved location in the acrosomal region.


Subject(s)
Acrosome/metabolism , Seminal Plasma Proteins/chemistry , Serine Endopeptidases/chemistry , Spermatozoa/metabolism , Animals , Blotting, Western , Cattle , Electrophoresis, Polyacrylamide Gel , Fluorescent Antibody Technique, Indirect , Gelatin/metabolism , Hydrolysis , Male , Microscopy, Fluorescence , RNA, Messenger/metabolism , Species Specificity
11.
Acta Otorrinolaringol Esp ; 55(5): 240-3, 2004 May.
Article in Spanish | MEDLINE | ID: mdl-15461322

ABSTRACT

Nasal tuberculosis represents a rare manifestation of infection by Mycobacterium tuberculosis. Clinically, it appeared to resemble cancer presenting as a nasopharyngeal mass with concomitant enlarged lymph nodes. It is important to consider tuberculosis in the differential diagnosis of all nasopharyngeal lesions and take biopsy samples for histological and bacteriological studies. Antituberculosis treatment is satisfactory with standard anti-tuberculous chemotherapy. Although this is a rare finding, it should be considered when a patient presents with nasal obstruction.


Subject(s)
Nasal Obstruction/etiology , Nasopharyngeal Diseases/diagnosis , Tuberculosis/diagnosis , Diagnosis, Differential , Female , Humans , Middle Aged , Nasopharyngeal Diseases/microbiology
12.
Acta Otorrinolaringol Esp ; 54(6): 465-9, 2003.
Article in Spanish | MEDLINE | ID: mdl-14567081

ABSTRACT

The Riedel's thyroiditis is a chronic inflammatory disease of the thyroid gland characterized by a process of fibrosis that destroys the gland partially, extending to the adjacent cervical structures. Its origin is unknown, although there are several theories in this respect. The clinical manifestations are not specific and it confuses frequently with neoplasia or infection, like a hard and painless thyroid mass that causes compressive symptoms. In this work, we report a case with lethal outcome.


Subject(s)
Thyroiditis/diagnostic imaging , Thyroiditis/pathology , Aged , Fatal Outcome , Female , Humans , Radiography, Thoracic/methods , Ultrasonography
13.
Acta Otorrinolaringol Esp ; 54(5): 384-7, 2003 May.
Article in Spanish | MEDLINE | ID: mdl-12916481

ABSTRACT

The entity known as sinus histiocytosis with massive lymphadenopathy (SHML), or Rosai-Dorfman disease, is an uncommon benign proliferation of hematopoietic and fibrous tissue is that often present in the head and neck region. Its initial manifestations include bilateral cervical adenopathies, and in a high percentage it has an extranodal affectation. Its cause is unknown and it usually resolves satisfactorily. We present a case and we revise the literature.


Subject(s)
Histiocytosis, Sinus/diagnostic imaging , Child , Female , Histiocytosis, Sinus/microbiology , Histiocytosis, Sinus/pathology , Humans , Neck , Staphylococcal Infections/microbiology , Staphylococcus aureus/isolation & purification , Tomography, X-Ray Computed
14.
J Androl ; 22(1): 96-103, 2001.
Article in English | MEDLINE | ID: mdl-11191093

ABSTRACT

The sperm acrosome is a uniquely regulated secretory vesicle containing several hydrolase enzymes, including acid phosphatase (AP). The exocytotic event that releases these enzymes, the acrosome reaction, is required for fertilization in mammals. Different methods have been described in the scientific literature for detection of the acrosome reaction: double and triple stains, fluorescent-lectin stains, monoclonal antibodies against acrosomal antigens (immunodetection techniques), Coomassie blue, differential interference contrast or phase contrast, flow cytometry, and chlortetracycline (CTC). In contrast, only 1 method to detect AP released by live and reacted sperm has been described in the literature thus far. In this work we compare 2 classical methods, CTC and transmission electron microscopy (TEM), with the assay of AP released from the acrosome. AP released during the acrosome reaction was measured in the culture medium. Enzyme remaining in nonreacted sperm cells was released by Triton X-100 treatment. This enzyme-based methodology shows an increase of AP in the culture media after the acrosome reaction and a corresponding decrease in the detergent-releasable enzyme. The AP assay thus permits the detection of the mouse acrosome reaction and compares well with the CTC and TEM methods. This method is performed on the whole sperm population and so avoids the observer error that is inherent in light microscopic methods.


Subject(s)
Acid Phosphatase/metabolism , Acrosome Reaction/physiology , Spermatozoa/enzymology , Spermatozoa/ultrastructure , Analysis of Variance , Animals , Anti-Bacterial Agents , Biomarkers , Chlortetracycline , Culture Media , Male , Mice , Microscopy, Electron
15.
Medifam (Madr.) ; 11(2): 55-64, feb. 2001. tab
Article in ES | IBECS | ID: ibc-11687

ABSTRACT

La Incontinencia Urinaria (IU) había sido definida como un síntoma, pero actualmente constituye un problema importante de salud, puesto que afecta a la calidad de vida de las personas y su impacto socioeconómico es elevado.Su prevalencia es difícil de precisar porque las personas que la padecen no consultan específicamente por este problema, bien por desconocimiento (no creen que tenga solución), o porque aprenden a vivir con la IU.La IU no es un problema sin tratamiento. Depende de las causas, porque hay formas transitorias de incontinencia que se resuelven una vez tratada la causa que las ha originado. Existen también formas establecidas de incontinencia que precisan estudio y tienen tratamiento. Alguno de estos tratamientos pueden ser prescritos y seguidos por el médico de Atención Primaria.En otras ocasiones el paciente deberá ser derivado para estudio, una vez valoradas sus características individuales, como estado clínico, situación funcional, impacto y posibilidades reales de mejorar este problema (AU)


Subject(s)
Urinary Incontinence/epidemiology , Primary Health Care , Quality of Life , Socioeconomic Factors
16.
Arch Androl ; 44(2): 85-91, 2000.
Article in English | MEDLINE | ID: mdl-10746864

ABSTRACT

Glycosidases in rat epididymal fluid are secreted under androgen stimulation and possess receptors on the sperm surface. One of these enzymes, beta-D-galactosidase (gal), was found in the epididymal fluid as a soluble enzyme and also in a heterogeneous population of membrane bound vesicles (mbv). beta-D-Galactosidase was specifically localized to a subpopulation of larger, electron-dense mbv. The aim of this study was to analyze the high-affinity sites for gal on the membrane of mbv using two different methods: classical fluorometric assay (used in previous papers) and colloidal gold (20 nm) conjugated to gal as a marker in ultrastructural studies. beta-D-Galactosidase bound to mbv with high-affinity (Kd in a nanomolar range) are in a saturable form. Furthermore, 25 mM fructose-1,6-diphosphate (f-1,6-dip), a sugar that competes for the binding site, showed 50% inhibition of the binding. The gold conjugates were mostly observed on the surface of the large, electron-dense mbv but not on the small, electron-lucent mbv. Gold particles were also observed on the larger vesicles, but less frequently in the presence of f-1.6-dip. Larger mbv possesses high-affinity sites for gal on their membrane.


Subject(s)
Epididymis/enzymology , beta-Galactosidase/metabolism , Animals , Body Fluids/enzymology , Cell Membrane/enzymology , Cell Membrane/ultrastructure , Fluorometry , Gold Colloid , Male , Organelles/enzymology , Organelles/ultrastructure , Rats , beta-Galactosidase/ultrastructure
17.
Dev Biol ; 214(2): 429-43, 1999 Oct 15.
Article in English | MEDLINE | ID: mdl-10525345

ABSTRACT

We previously demonstrated that mouse sperm capacitation is accompanied by a time-dependent increase in protein tyrosine phosphorylation that is dependent on the presence of BSA, Ca2+, and NaHCO(3), all three of which are also required for this maturational event. We also demonstrated that activation of protein kinase A (PK-A) is upstream of this capacitation-associated increase in protein tyrosine phosphorylation. BSA is hypothesized to modulate capacitation through the removal of cholesterol from the sperm plasma membrane. In this report, we demonstrate that incubation of mouse sperm medium containing BSA results in a release of cholesterol from the sperm plasma membrane to the medium; release of this sterol does not occur in medium devoid of BSA. We next determined whether cholesterol release leads to changes in protein tyrosine phosphorylation. Blocking the action of BSA by adding exogenous cholesterol-SO-(4) to the BSA-containing medium inhibits the increase in protein tyrosine phosphorylation as well as capacitation. This inhibitory effect is overcome by (1) the addition of increasing concentrations of BSA at a given concentration of cholesterol-SO-(4) and (2) the addition of dibutyryl cAMP plus IBMX. High-density lipoprotein (HDL), another cholesterol binding protein, also supports the capacitation-associated increase in protein tyrosine phosphorylation through a cAMP-dependent pathway, whereas proteins that do not interact with cholesterol have no effect. HDL also supports sperm capacitation, as assessed by fertilization in vitro. Finally, we previously demonstrated that HCO-(3) is necessary for the capacitation-associated increase in protein tyrosine phosphorylation and demonstrate here, by examining the effectiveness of HCO-(3) or BSA addition to sperm on protein tyrosine phosphorylation, that the HCO-(3) effect is downstream of the site of BSA action. Taken together, these data demonstrate that cholesterol release is associated with the activation of a transmembrane signal transduction pathway involving PK-A and protein tyrosine phosphorylation, leading to functional maturation of the sperm.


Subject(s)
Cholesterol/metabolism , Signal Transduction , Sperm Capacitation/physiology , Spermatozoa/metabolism , Acrosome/metabolism , Animals , Cholesterol Esters/pharmacology , Cyclic AMP/agonists , Cyclic AMP/metabolism , Cyclic AMP-Dependent Protein Kinases/metabolism , Desmosterol/metabolism , Dose-Response Relationship, Drug , Fertilization , Filipin/metabolism , Freeze Fracturing , Lipoproteins, HDL/pharmacology , Male , Mice , Phosphorylation , Serum Albumin, Bovine/pharmacology , Sterols/metabolism , Time Factors , Tyrosine/metabolism
18.
Mol Reprod Dev ; 52(3): 297-302, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10206661

ABSTRACT

G-proteins, calcium, and phospholipase A2 (PLA2) have all been implicated in the cascade of signaling events leading to the acrosome reaction in human spermatozoa. In order to study the role of Ca+2 and PLA2 during the acrosome reaction triggered by G-proteins, we treated human spermatozoa incubated for 3 hr under capacitating conditions with several reagents (GTPgammaS, A23187, ONO-RS-082, arachidonic acid, BAPTA-AM, and TPEN), alone or in different combinations. Our results suggest that GTP-binding proteins require Ca+2 and PLA2 to accomplish their stimulatory effect, and that Ca+2 is also required when the acrosome reaction--bypassing the action of PLA2--is stimulated by AA. Accordingly, when treated with GTPgammaS or AA, the cells loaded with Fura 2-AM showed a steady increase of [Ca+2]i. On the other hand, a massive influx of Ca+2 was completely unable to induce the acrosome reaction if PLA2 was inhibited, suggesting that both an increase of [Ca+2]i and PLA2 activation are required for the acrosome reaction to occur.


Subject(s)
Acrosome Reaction/physiology , Calcium/metabolism , GTP-Binding Proteins/metabolism , Phospholipases A/metabolism , Spermatozoa/physiology , Acrosome Reaction/drug effects , Arachidonic Acid/metabolism , Calcimycin/metabolism , Calcimycin/pharmacology , Guanosine 5'-O-(3-Thiotriphosphate)/metabolism , Humans , Ionophores/metabolism , Ionophores/pharmacology , Male , Phospholipases A2 , Spermatozoa/drug effects
19.
Andrologia ; 31(1): 23-6, 1999 Jan.
Article in English | MEDLINE | ID: mdl-9949885

ABSTRACT

Because morphology is regularly established in semen smears, but not in swim-up spermatozoa, we were interested in comparing some morphological parameters of semen and swim-up spermatozoa to establish if the cells selected by the swim-up method were morphologically similar to those considered normal in semen. Normal human semen samples were divided into two aliquots. One of these aliquots was washed by centrifugation with B2 medium and sperm smears were prepared with the resulting pellet as a control. The other aliquot was used to perform swim-up separation and the spermatozoa from the supernatant were used as experimental smears. Both groups were stained according to the triple stain technique and spontaneous acrosome reaction and viability were determined. Video microscopy and computer-assisted image processing of live and non-reacted sperm cells were used to establish morphometrical parameters of the sperm head in both populations. The following set of morphometrical parameters were considered: width, length, width/length ratio, acrosome area, head area, and acrosome area/head area ratio. An increase in head width, a decrease in head length and a subsequent increase of width/length ratio were found in swim-up cells compared with the control group. A slight increase in acrosome area/head area ratio was also observed in swim-up supermatozoa. Through the swim-up methodology we were able to select a subpopulation of oval shaped heads with spermatozoa having a bigger acrosome area in comparison to semen.


Subject(s)
Semen/cytology , Sperm Motility , Spermatozoa/cytology , Acrosome , Humans , Male
20.
Actas Urol Esp ; 22(6): 485-9, 1998 Jun.
Article in Spanish | MEDLINE | ID: mdl-9734124

ABSTRACT

Transurethral resection (TUR) is the most frequent surgical treatment for symptomatic benign prostate hyperplasia (BPH). Prostate size is a significant factor for choosing TUR versus prostate adenomectomy. Analysis of flowmetry results obtained with TUR in 203 patients, based on weight of resected prostate tissue. Flowmetry was performed prior to TUR and prostate size was estimated with transabdominal ultrasound. Prostate tissue was weighed after TUR and a new flowmetry was performed 6 months after treatment. Mean weight of resected tissue was 31.34 g. When all flowmetry parameters analyzed pre-and post TURs were compare, there were significant differences (p < 0.001). Increased maximum flow (Qmax) and increased mean flow (Qmed) occurred in 91.7% and 96.31% patients, respectively. Mean increase of post-surgical Qmax was significantly higher (p > 0.01) in patients with Qmax prior to surgery lower than 8 ml/s. No significant correlation was demonstrated between prostate volume measured by ultrasound or resected prostate tissue and increased post-surgical Qmax. TUR improves flowmetry parameters, mainly in patients with pre-surgical Qmax lower than 8 mL/s. Extensive prostate resection does not appear to improve the flowmetry results obtained with a sufficient functional TUR.


Subject(s)
Prostatectomy , Urination , Aged , Humans , Male , Rheology , Urine
SELECTION OF CITATIONS
SEARCH DETAIL
...