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1.
Reprod Biol Endocrinol ; 10: 81, 2012 Sep 12.
Article in English | MEDLINE | ID: mdl-22971106

ABSTRACT

BACKGROUND: The use of therapeutic ultrasound as a contraceptive approach has involved nonhuman primates as well as rats and dogs. The current study was undertaken to determine whether this treatment could be a method for reversible contraception, using a model with testes size similar to adult humans. METHODS: Two methods of ultrasound exposure were used, either the transducer probe at the bottom of a cup filled with saline (Cup) or direct application to the surface of the scrotum (Direct). Four adult rhesus (Macaca mulatta) males with normal semen parameters were treated with therapeutic ultrasound at 2.5 W/cm(2) for 30 min. Treatment was given 3 times, one every other day on a Monday-Wednesday-Friday schedule. For each male, semen quality was evaluated a minimum of three times over several months prior to ultrasound exposure and weekly for two months following ultrasound treatment. RESULTS: Semen samples from all males, regardless of exposure method, exhibited a decrease in the percentage of motile sperm following ultrasound treatment. There was an average reduction in motility of 40% the week following treatment. Similarly, curvilinear velocity and the percentage of sperm with a normally shaped flagellum were also reduced in all males following ultrasound treatment. A significant reduction in the total number of sperm in an ejaculate (total sperm count) was only observed in males that received ultrasound via the cup method. Following treatment via the cup method, males exhibited up to a 91.7% decrease in average total sperm count (n = 2). Sperm count did not approach pre-treatment levels until 8 weeks following ultrasound exposure. CONCLUSIONS: The sustained reduction in sperm count, percent motility, normal morphology, and sperm vigor with the cup exposure method provides proof of principle that testicular treatment with ultrasound can be an effective contraceptive approach in humans.


Subject(s)
Contraception/methods , Spermatozoa/diagnostic imaging , Testis/diagnostic imaging , Animals , Contraception/adverse effects , Contraception/instrumentation , Indicators and Reagents/chemistry , Macaca mulatta , Male , Scrotum/diagnostic imaging , Semen Analysis , Sodium Chloride/chemistry , Sperm Tail/diagnostic imaging , Spermatogenesis , Spermatozoa/cytology , Time Factors , Ultrasonography
2.
Hum Reprod ; 24(6): 1288-95, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19221095

ABSTRACT

BACKGROUND: There is no systematic study on coiled sperm in semen, although they are commonly observed. This work characterizes coiled sperm in infertile men to understand the clinical implications and investigate the possible cause by osmotic swelling. METHODS: Coiled sperm in semen from 439 infertile patients were quantified and their ultrastructure examined by electron microscopy. Hypo-osmotic swelling (HOS) and demembranation tests were performed to elucidate the nature of the coiling. RESULTS: Semen from patients contained overall 3% of sperm with head-in-coil (HIC) and 8% other coiled forms, with 12% of patients having 20% or more such sperm. The percentage of coiled sperm (but not HIC) was correlated with age (R = 0.26, P = 0.003) and the epididymal secretory marker neutral alpha-glucosidase (R = 0.16, P < 0.001), and associated with heavy smoking and varicocele. Electron microscopy revealed coiling of tail filaments within the plasma membrane, resembling HOS. Some seminal coiled sperm and most sperm freshly coiled upon HOS could be opened by demembranation, while those that could not be opened were probably fixed in position by oxidation, which occurred more frequently in patients than semen donors. CONCLUSIONS: Sperm coiling in semen is common and independent of sperm quantity or hormonal status. Whereas HIC may have a genetic background, other coiled forms may be associated with a hostile endogenous milieu in the epididymis that causes swelling.


Subject(s)
Infertility, Male/pathology , Sperm Tail/diagnostic imaging , Sperm Tail/pathology , Varicocele/pathology , Adolescent , Adult , Cell Membrane/diagnostic imaging , Cell Membrane/metabolism , Cell Membrane/pathology , Hormones/blood , Humans , Infertility, Male/metabolism , Male , Microscopy, Electron , Middle Aged , Osmolar Concentration , Osmosis , Smoking , Sperm Tail/metabolism , Ultrasonography , Varicocele/metabolism , Young Adult
3.
Theriogenology ; 62(7): 1307-28, 2004 Oct 01.
Article in English | MEDLINE | ID: mdl-15325557

ABSTRACT

The objectives of this study were to describe the features of experimentally induced orchitis associated with Arcanobacterium pyogenes and confirm the pathogenicity of the organism for the ovine testicle. One testicle of each of nine rams was inoculated with 1.3 +/- 10(4) colony-forming-units of an A. pyogenes isolate and regular clinical, ultrasonographic, bacteriological and seminological examinations were carried out up to 204 days after challenge. The rams were sequentially euthanatized 3, 6, 9, 18, 30, 50, 71, 113 and 204 days after challenge and a gross- and histopathological examination of their testicles was performed. All rams developed clinical orchitis and general signs. The initial ultrasonographic findings were changes of size and echogenicity of the genitalia, whilst in the long-standing phase they were wider appearance of the mediastinum testis, presence of hyperechogenic foci, changes of echogenicity of the genitalia and increased echogenicity of the scrotum and tunics. The following changes in semen evaluation parametres were recorded: the pH, the percentage of dead sperms, the percentage of abnormal sperms and the number of nonsperm round cells increased, whilst the mass motility, the individual motility and the sperm concentration decreased; the following sperm defects were observed: misshapen or piriform heads, sperms with coiled tails, sperms without tail and sperms with proximal cytoplasmic droplet; at the early stages neutrophils were the prevailing nonsperm round cell type, later the proportion of immature germ cells increased and in the long-standing phase there were enlogated spermatids and leucocytes; it is noteworthy that semen evaluation parametres were restored to normal at the late stages of the disease. A. pyogenes was consistently isolated from the semen samples after challenge, as well as from the dissected genitalia. The salient post-mortem findings were: initially, subcutaneous oedema, fluid into the vaginal cavity, congested and distended vessels, increased size of the genitalia and a hard dark area inside the testicles; subsequently, there were changes of size of the genitalia, thickening of scrotum and tunics and presence of fibrin on the testicular surface; in the long-standing phase of the disorder, there were induration of scrotum and tunics with adhesion between the tunics and discolouration of the surface of the genitalia. The prominent histopathological changes were observed in the inoculated testicles; milder changes were seen in the respective epididymides; interstitial oedema, diffuse neutrophilic infiltration and extravasation were observed in the early stages after challenge; lymphocytic infiltration with concurrent fibrosis, mineralization and inspissation of the tubular elements of the seminiferous tubules and presence of vacuolated Sertoli cells were seen later; finally, regeneration of the epithelium and presence of Sertoli cells and spermatogonia with various degrees of spermatogenic activity were evident. These findings, allied to the isolation of A. pyogenes from field cases of ovine orchitis, provide clear evidence that A. pyogenes is pathogenic for the ovine genitalia; however, the mechanisms of transition of the organism from commensal to pathogenic state are not clear. It is also noteworthy that some degree of fertility was restored in the late stages of the disorder. Ultrasonography appeared to be useful for the diagnosis of intra-scrotal abnormalities, especially during investigation of the long-standing stage of the disease, after clinical findings have subsided.


Subject(s)
Corynebacterium Infections/veterinary , Corynebacterium pyogenes , Epididymis/pathology , Orchitis/veterinary , Sheep Diseases/physiopathology , Spermatozoa/pathology , Testis/pathology , Acrosome/diagnostic imaging , Acrosome/pathology , Animals , Corynebacterium Infections/diagnostic imaging , Corynebacterium Infections/physiopathology , Epididymis/diagnostic imaging , Male , Orchitis/diagnostic imaging , Orchitis/physiopathology , Random Allocation , Sheep , Sheep Diseases/diagnostic imaging , Sheep Diseases/microbiology , Sperm Count/veterinary , Sperm Head/diagnostic imaging , Sperm Head/pathology , Sperm Motility , Sperm Tail/diagnostic imaging , Sperm Tail/pathology , Spermatozoa/diagnostic imaging , Testis/diagnostic imaging , Ultrasonography
4.
Dev Biol ; 270(2): 290-307, 2004 Jun 15.
Article in English | MEDLINE | ID: mdl-15183715

ABSTRACT

Macroorchidism (i.e., enlarged testicles) and mental retardation are the two hallmark symptoms of Fragile X syndrome (FraX). The disease is caused by loss of fragile X mental retardation protein (FMRP), an RNA-binding translational regulator. We previously established a FraX model in Drosophila, showing that the fly FMRP homologue, dFXR, acts as a negative translational regulator of microtubule-associated Futsch to control stability of the microtubule cytoskeleton during nervous system development. Here, we investigate dFXR function in the testes. Male dfxr null mutants have the enlarged testes characteristic of the disease and are nearly sterile (>90% reduced male fecundity). dFXR protein is highly enriched in Drosophila testes, particularly in spermatogenic cells during the early stages of spermatogenesis. Cytological analyses reveal that spermatogenesis is arrested specifically in late-stage spermatid differentiation following individualization. Ultrastructurally, dfxr mutants lose specifically the central pair microtubules in the sperm tail axoneme. The frequency of central pair microtubule loss becomes progressively greater as spermatogenesis progresses, suggesting that dFXR regulates microtubule stability. Proteomic analyses reveal that chaperones Hsp60B-, Hsp68-, Hsp90-related protein TRAP1, and other proteins have altered expression in dfxr mutant testes. Taken together with our previous nervous system results, these data suggest a common model in which dFXR regulates microtubule stability in both synaptogenesis in the nervous system and spermatogenesis in the testes. The characterization of dfxr function in the testes paves the way to genetic screens for modifiers of dfxr-induced male sterility, as a means to efficiently dissect FMRP-mediated mechanisms.


Subject(s)
Drosophila Proteins/genetics , Drosophila/genetics , Fragile X Syndrome/genetics , Microtubules/physiology , RNA-Binding Proteins/genetics , Spermatogenesis/physiology , Testis/physiology , Animals , Blotting, Western , Disease Models, Animal , Drosophila/physiology , Drosophila Proteins/metabolism , Drosophila Proteins/physiology , Electrophoresis, Gel, Two-Dimensional , Electrophoresis, Polyacrylamide Gel , Fragile X Mental Retardation Protein , Immunohistochemistry , Male , Microscopy, Confocal , Microscopy, Electron , Microtubules/diagnostic imaging , Molecular Chaperones/metabolism , RNA-Binding Proteins/metabolism , RNA-Binding Proteins/physiology , Sperm Tail/diagnostic imaging , Spermatogenesis/genetics , Testis/metabolism , Testis/ultrastructure , Ultrasonography
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