Sperm Motility Defects Research Articles

A Rare Case of Respiratory Disorders Associated with Two Autosomal Recessive Diseases and Male Infertility

The study of nasal ciliary beat frequency (CBF) and ultrastructure may contribute to the understanding of pathognomonic cases of male infertility associated with defects in sperm motility. This study was designed to report a particular case of male infertility, characterized by the association of two respiratory autosomal recessive genetic diseases (alpha-1-antitrypsin deficiency [AAT-D] and primary ciliary dyskinesia [PCD]). A 39-year-old patient with complete sperm immotility, AAT-D, and bronchiectasis was studied in the Laboratory of Male Fertility, the Department of Urology, the Respiratory Center of a Pediatric Hospital, and in the Department of Clinical Medicine of a Rehabilitation Respiratory Hospital. Family history, physical examination, hormonal analysis, microbial assays, semen analysis, nasal ciliary function, and structure study by digital high-speed video photography and transmission electron microscopy are described. A noninvasive nasal biopsy to retrieve ciliated epithelium lining the inferior surface of the inferior nasal turbinates was performed and CBF was determined. Beat pattern was slightly curved and rigid, not wide, and metacronic in all the observed fields analyzed. CBF was 8.2 Hz in average (reference value, 10–15 Hz) Ultrastructural assay revealed absence of the inner dynein arms in 97% of the cilia observed. The final infertility accurate diagnosis was achieved by the study of nasal CBF and ultrastructure contributing to the patient health management and genetic counseling while deciding fatherhood. Beyond this particular case, the present report may open a new field of studies in male infertility, mainly in cases of asthenozoospermia.

Genetic variation in SPAG16 regions encoding the WD40 repeats is not associated with reduced sperm motility and axonemal defects in a population of infertile males

BackgroundSPAG16 is a critical structural component of motile cilia and flagella. In the eukaryotic unicellular algae Chlamydomonas, loss of gene function causes flagellar paralysis and prevents assembly of the “9 + 2” axoneme central pair. In mice, we have previously shown that loss of Spag16 gene function causes male infertility and severe sperm motility defects. We have also reported that a heterozygous mutation of the human SPAG16 gene reduces stability of the sperm axonemal central apparatus.MethodsIn the present study, we analyzed DNA samples from 60 infertile male volunteers of Western European (Italian) origin, to search for novel SPAG16 gene mutations, and to determine whether increased prevalence of SPAG16 single nucleotide polymorphisms (SNPs) was associated with infertility phenotypes. Semen parameters were evaluated by light microscopy and sperm morphology was comprehensively analyzed by transmission electron microscopy (TEM).ResultsFor gene analysis, sequences were generated covering exons encoding the conserved WD40 repeat region of the SPAG16 protein and the flanking splice junctions. No novel mutations were found, and the four SNPs in the assessed gene region were present at expected frequencies. The minor alleles were not associated with any assessed sperm parameter in the sample population.ConclusionsAnalysis of the SPAG16 regions encoding the conserved WD repeats revealed no evidence for association of mutations or genetic variation with sperm motility and ultrastructural sperm characteristics in a cohort of Italian infertile males.

Asymptomatic seminal infection of herpes simplex virus

In more than half of infertile men, the cause of their infertility is unknown. Several studies revealed the role of viral infections in male infertility. The aim of the present study was to determine the prevalence of herpes simplex virus-1 (HSV-1) and HSV-2 in semen from asymptomatic infertile male patients, and its association with altered semen parameters. A total of 70 semen samples were collected from infertile men who attended the Research and Clinical Center for Infertility in Yazd, Iran. Semen analysis and diagnostic real-time PCR using specific primers and probes for HSV-1 and HSV-2 DNA were performed. Comparison of semen parameters between virally infected and non-infected samples were performed with independent t-test and Mann-Whitney test. Semen analysis showed that infertile men fell into two groups, the male factor group and the unexplained group. HSV-1 and HSV-2 DNA was detected in 16 (22.9%) and 10 (14.3%) of 70 semen samples, respectively. All HSV-positive samples had abnormal semen parameters (the male factor group). Although HSV infection was not associated with sperm motility and morphological defects, it was correlated with lower sperm count in the seminal fluid. The findings suggest that asymptomatic seminal infection of HSV plays an important role in male infertility by adversely affecting sperm count.

Field Data Analysis of Boar Semen Quality

This contribution provides an overview of approaches to correlate sow fertility data with boar semen quality characteristics. Large data sets of fertility data and ejaculate data are more suitable to analyse effects of semen quality characteristics on field fertility. Variation in fertility in sows is large. The effect of semen factors is relatively small and therefore impossible to find in smaller data sets. Large data sets allow for statistical corrections on both sow- and boar-related parameters. Remaining sow fertility variation can then be assigned to semen quality parameters, which is of huge interest to AI (artificial insemination) companies. Previous studies of Varkens KI Nederland to find the contribution to field fertility of (i) the number of sperm cells in an insemination dose, (ii) the sperm motility and morphological defects and (iii) the age of semen at the moment of insemination are discussed in context of the possibility to apply such knowledge to select boars on the basis of their sperm parameters for AI purposes.

A comprehensive work up for an asthenozoospermic man with repeated intracytoplasmic sperm injection (ICSI) failure

Infertility affects about 15-20% couples attempting pregnancy and in about half cases the problem lies in the male. Among the sperm parameters, linear progressive motility is one of the most important predictors of fertility potential. Though genetic and chromosomal abnormalities are important aetiological factors in the pathogenesis of male infertility, the mechanism involved in impaired sperm motility is poorly understood. Here we report mitochondrial DNA (mtDNA) mutations with increased seminal reactive oxygen species (ROS) levels and higher DNA fragmentation level in the sperm resulting in decreased ATP production which plays an important role in sperm motility defect. Thus it is important to understand the aetiology of asthenozoospermia and to distinguish if infertile men harbour nuclear or mtDNA mutation as they are very important prognostic markers. This case study also highlights that routine semen parameters are very modest predictors of fertility outcome but ROS estimation and DNA integrity analysis by Comet assay have better diagnostic and prognostic capabilities. Thus this study is a detailed and comprehensive workup of an infertile asthenozoospermic male.

Mice Lacking the USP2 Deubiquitinating Enzyme Have Severe Male Subfertility Associated with Defects in Fertilization and Sperm Motility

The ubiquitin-proteasome system plays an important role in spermatogenesis. However, the functions of deubiquitinating enzymes in this process remain poorly characterized. We previously showed that the deubiquitinating enzyme USP2 is induced in late elongating spermatids. To identify its function, we generated mice lacking USP2. Usp2 -/- mice appeared normal, and the weights of major organs, including the testis, did not differ from wild type (Usp2 +/+). However, although the numbers of testicular spermatids and epididymal spermatozoa were normal in Usp2 -/- males, these animals had a severe defect in fertility, yielding only 12% as many offspring as Usp2 +/+ littermates. Spermatogenesis in Usp2 -/- mice was morphologically normal except for the presence of abnormal aggregations of elongating spermatids and formation of multinucleated cells in some tubules. The epididymal epithelium was morphologically normal in Usp2 -/- mice, but some abnormal cells other than sperm were present in the lumen. Usp2 -/- epididymal spermatozoa manifested normal motility when incubated in culture media, but rapidly became immotile when incubated in PBS in contrast to Usp2 +/+ spermatozoa, which largely maintained motility under this condition. Usp2 -/- and +/+ spermatozoa underwent acrosome reactions in vitro with similar frequency. In vitro fertilization assays demonstrated a severe defect in the ability of Usp2 -/- spermatozoa to fertilize eggs. This could be bypassed by intracytoplasmic sperm injection or removal of the zona pellucida, which resulted in fertilization rates similar to that of Usp2 +/+ mice. We demonstrate for the first time, using mouse transgenic approaches, a role for the ubiquitin system in fertilization.

Lack of Tyrosylprotein Sulfotransferase-2 Activity Results in Altered Sperm-Egg Interactions and Loss of ADAM3 and ADAM6 in Epididymal Sperm

Tyrosine O-sulfation is a post-translational modification catalyzed by two tyrosylprotein sulfotransferases (TPST-1 and TPST-2) in the trans-Golgi network. Tpst2-deficient mice have male infertility, sperm motility defects, and possible abnormalities in sperm-egg membrane interactions. Studies here show that compared with wild-type sperm, fewer Tpst2-null sperm bind to the egg membrane, but more of these bound sperm progress to membrane fusion. Similar outcomes were observed with wild-type sperm treated with the anti-sulfotyrosine antibody PSG2. The increased extent of sperm-egg fusion is not due to a failure of Tpst2-null sperm to trigger establishment of the egg membrane block to polyspermy. Anti-sulfotyrosine staining of sperm showed localization similar to that of IZUMO1, a sperm protein that is essential for gamete fusion, but we detected little to no tyrosine sulfation of IZUMO1 and found that IZUMO1 expression and localization were normal in Tpst2-null sperm. Turning to a discovery-driven approach, we used mass spectrometry to characterize sperm proteins that associated with PSG2. This identified ADAM6, a member of the A disintegrin and A metalloprotease (ADAM) family; members of this protein family are associated with multiple sperm functions. Subsequent studies revealed that Tpst2-null sperm lack ADAM6 and ADAM3. Loss of ADAM3 is strongly associated with male infertility and is observed in knockouts of male germ line-specific endoplasmic reticulum-resident chaperones, raising the possibility that TPST-2 may function in quality control in the secretory pathway. These data suggest that TPST-2-mediated tyrosine O-sulfation participates in regulating the sperm surface proteome or membrane order, ultimately affecting male fertility.

Effect of ascorbic acid on chicken semen quality during liquid storage

The aim of this study was to evaluate the in vitro effects of ascorbic acid on Ross-308 chicken sperm quality. The semen was collected from eight teen Ross-308 broiler breeder roosters and mixed together. The mixed semen was diluted with modified Ringer’s solution and supplemented with five treatment levels (0%, 0.5%, 1%, 2%, and 3%) of ascorbic acid (w/v). Treatments were evaluated for sperm motility, viability, and morphological defect rates at 0, 3, 6, 10, and 24 h after storage at 4°C. At 0, 3, and 6 of storage, no significant differences were observed between treatments with ascorbic acid and controls for all spermatozoa quality parameters. At 10 h, sperm viability in treatment added with 1% ascorbic acid was significantly higher than control and other treatments. In this time, there were not significant differences among treatments with 0%, 0.5%, 2%, and 3% ascorbic acid. At 24 h, sperm motility in treatment with 1% ascorbic acid and sperm viability in treatments contain 1% and 2% ascorbic acid were higher than controls and other treatments. In this period, morphological defect percent of spermatozoa in treatment with 1% ascorbic acid was significantly lower than control and other treatment groups. Also, at 24 h, sperm morphological defect percent in treatment added with 3% ascorbic acid was significantly higher than that in control and other treatments.

Role of a-kinase anchoring proteins (AKAPs) in reproduction

Rapid spatio-temporal organized intracellular signaling is a pivotal mechanism for regulation of functions in many cells, in particular in the female and male gametes, in which functional regulation through rapid increases in protein content is not possible since the mechanisms of transcription/translation are somehow frozen due to meiosis block or DNA condensation respectively. A kinase-anchoring proteins (AKAPs) represent a functional conserved family of signal-organizing scaffolding proteins, which due to the specific subcellular distribution and focally compartmentalized cyclic-AMP-dependent protein kinase (PKA) and other enzymes, assuring the coordination of cAMP-responsive events and their integration with other intracellular signals. This review summarizes the actual knowledge on AKAP structure and functions, taking into particular account the role of different AKAPs in regulating reproductive functions such as gametogenesis. Evidence for sperm specific AKAP isoforms and their initiated signaling cascades in mature sperm and the role of this focally activated super-molecular signaling complex in motility are discussed in details with particular emphasis on putative relations between AKAP structural and functional alterations and defects in sperm motility.

Insulin/FOXO Signaling Regulates Ovarian Prostaglandins Critical for Reproduction

Abnormalities in insulin/IGF-1 signaling are associated with infertility, but the molecular mechanisms are not well understood. Here we use liquid chromatography with electrospray ionization tandem mass spectrometry to show that the C. elegans insulin/FOXO pathway regulates the metabolism of locally acting lipid hormones called prostaglandins. C. elegans prostaglandins are synthesized without prostaglandin G/H synthase homologs, the targets of nonsteroidal anti-inflammatory drugs. Our results support the model that insulin signaling promotes the conversion of oocyte polyunsaturated fatty acids (PUFAs) into F-series prostaglandins that guide sperm to the fertilization site. Reduction in insulin signaling activates DAF-16/FOXO, which represses the transcription of germline and intestinal genes required to deliver PUFAs to oocytes in lipoprotein complexes. Nutritional and neuroendocrine cues target this mechanism to control prostaglandin metabolism and reproductive output. Prostaglandins may be conserved sperm guidance factors and widespread downstream effectors of insulin actions that influence both reproductive and nonreproductive processes.

A Multiclass Support Vector Machine Model for the Identification of Sperm Motility Patterns in Heterogeneous Populations.

Sperm motility and the capacity to undergo hyperactivation in the female reproductive tract are requirements for normal fertilization in mammals. Quantitative analyses of motility in response to perturbations that modulate sperm metabolism or key signaling events would be facilitated by a method that quickly and accurately distinguishes different types of motility in large populations of sperm. To that end, we generated a multiclass support vector machine (SVM) model that identifies hyperactivated sperm and four other distinct patterns of sperm motility within a heterogeneous population. Our multiclass model is based on 2,434 sperm tracks from twelve CD1 mice that were captured using a Hamilton-Thorne CEROS instrument for computer-assisted sperm analysis (CASA) and visually classified into one of six distinct motility groups: progressive, intermediate, hyperactivated, slow, weakly motile, and bad (noise). The CASA parameters from these classified tracks were used to develop a series of SVM equations using established algorithms, yielding a model that identifies uncharacterized tracks with an overall accuracy of 86%. The model incorporates these equations into a binary decision tree that sequentially sorts tracks into distinct populations. Five standard CASA measurements are factored into each equation, including average path velocity (VAP), straight-line velocity (VSL), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF). The first equation identifies all CASA tracks as either vigorous or non-vigorous. Vigorous tracks are then classified as progressive, intermediate or hyperactivated, while non-vigorous tracks are classified as slow, weakly motile, or bad. We tested our multiclass model by comparing motility profiles of sperm incubated for 2 h in HTF medium with and without bicarbonate. After capacitation for 90 min in the presence of bicarbonate, 20-30% of sperm tracks were classified as hyperactivated and ~10% as intermediate. Without bicarbonate, >65% of sperm tracks remained progressive throughout the incubation and <2% were classified as hyperactivated or intermediate, as expected. The model also accurately categorized tracks from mice with defects in sperm motility. For example, sperm from mice that lack GAPDHS, an essential sperm glycolytic isozyme, showed no progressive motility and >96% were classified as slow or weakly motile. We also assessed the motility profiles of C57BL6/J, 129S1/SvlmJ, and PWK/PhJ sperm over a 2 h in vitro capacitation period and found significant differences among these strains, both in the proportions of progressive and slow sperm and in the percentage of sperm that undergo hyperactivation. By enabling detailed, quantitative comparisons of motility throughout capacitation, this model will be a valuable tool for assessing potential genetic, biomolecular, and pharmaceutical effects on sperm motility in large, heterogeneous populations. Supported by the Eunice Kennedy Shriver NICHD/NIH through U01 HD60481 and cooperative agreement U54 HD35041 as part of the Specialized Cooperative Centers Program in Reproduction and Infertility Research. (platform)

The CIL-1 PI 5-Phosphatase Localizes TRP Polycystins to Cilia and Activates Sperm in C. elegans

C. elegans male sexual behaviors include chemotaxis and response to hermaphrodites, backing, turning, vulva location, spicule insertion, and sperm transfer, culminating in cross-fertilization of hermaphrodite oocytes with male sperm. The LOV-1 and PKD-2 transient receptor potential polycystin (TRPP) complex localizes to ciliated endings of C. elegans male-specific sensory neurons and mediates several aspects of male mating behavior. TRPP complex ciliary localization and sensory function are evolutionarily conserved. A genetic screen for C. elegans mutants with PKD-2 ciliary localization (Cil) defects led to the isolation of a mutation in the cil-1 gene. Here, we report that a phosphoinositide (PI) 5-phosphatase, CIL-1, regulates TRPP complex ciliary receptor localization and sperm activation. cil-1 does not regulate the localization of other ciliary proteins, including intraflagellar transport (IFT) components, sensory receptors, or other TRP channels in different cell types. Rather, cil-1 specifically controls TRPP complex trafficking in male-specific sensory neurons and does so in a cell-autonomous fashion. In these cells, cil-1 is required for normal PI(3)P distribution, indicating that a balance between PI(3,5)P2 and PI(3)P is important for TRPP localization. cil-1 mutants are infertile because of sperm activation and motility defects. In sperm, the CIL-1 5-phosphatase and a wortmannin-sensitive PI 3-kinase act antagonistically to regulate the conversion of sessile spermatids into motile spermatozoa, implicating PI(3,4,5)P3 signaling in nematode sperm activation. Our studies identify the CIL-1 5-phosphatase as a key regulator of PI metabolism in cell types that are important in several aspects of male reproductive biology.

The Effect of Season on Semen Characteristics and Freezability in Bos indicus and Bos taurus Bulls in the Southeastern Region of Brazil

The objectives of this study were to evaluate the effects of season in southeast of Brazil comparing genotypes on semen characteristics, freezability and peripheral plasma concentrations of testosterone. Ejaculates of five Bos indicus bulls and six Bos taurus bulls were evaluated over a period of 27 months, which was divided into winter (July, August, September), spring (October, November, December), summer (January, February, March) and autumn (April, May, June). Semen was evaluated according to standard procedures for ejaculate volume, sperm concentration, gross-motility, progressive motility and sperm morphology. After preparing and freezing the ejaculates according to commercial procedures, the straws were stored in liquid N(2) until post-thaw evaluation. Ejaculate volume, sperm concentration, gross-motility, progressive sperm motility, vigor and morphological sperm defects were significantly influenced by season and genotype (p < 0.05). Heat tolerance was better in B. indicus bulls than in B. taurus bulls characterized by lower values of sperm abnormalities throughout the observation period. The highest values were recorded for abnormal heads followed by cytoplasmatic droplets in B. taurus bulls. The proportion of ejaculates which were eliminated before freezing for reasons of bad quality was lower in the B. indicus bulls. Temporal changes in peripheral plasma testosterone concentrations were higher in B. indicus bulls than in B. taurus bulls not revealing seasonal influences. The results of this study show clear genotype differences regarding semen quality. Freezability of B. taurus semen varies considerably throughout the year, leading to a high proportion of eliminated ejaculates. Collecting semen from B. taurus bulls during the summer in an artificial insemination centre may not be profitable.

Oxidative stress and epididymal sperm transport, motility and morphological defects

Objectives Radical oxidative species (ROS) have an important effect on sperm quality and quantity. Oxidative stress (OS) occurs when production of potentially destructive reactive oxygen species (ROS) exceeds the body's own natural antioxidant defenses, resulting in cellular damage. OS is a common pathology seen in approximately half of all infertile men. Increased ROS generation and reduced antioxidant capacity is negatively correlated with sperm concentration and motility in infertile men. For the first time, we used a more stable and reliable sensitive carbonyl protein (CP) detection method to determine ROS in seminal plasma than measuring ROS directly to clarify the effect of OS on spermatozoa in terms of protein dysfunction. This is the first report to measure CP in seminal plasma as an indicator of OS. Furthermore, for the first time we correlated the results of CP measurement with light microscopy in combination with ultrastructural analysis by electron microscopy. Material and Methods 20 patients with idiopathic oligoasthenoteratozoospermia (iOAT) and 10 fertile controls were enrolled in this study. CP values were measured by enzyme-linked immuno sorbent assay (ELISA) to detect the level of OS. Transmission electron microscope (TEM) was used to detect axonemal anomalies. Results Compared to fertile controls, statistically highly significant higher degrees of abnormal sperm parameters ( P < 0.001) could be found in iOAT patients. CP values were highly significantly elevated in iOAT patients than in normal controls ( P < 0.001). A statistically highly significant difference in different axonemal anomalies were found between iOAT patients and normal controls ( P < 0.001). CP values have been found to be positively correlated with different axonemal anomalies (absence of axoneme ( r 2 = 0.841), missing of central singlet tubules ( r 2 = 0.702) and missing of outer doublet tubules ( r 2 = 0.869). A statistically negative correlation were found between different axonemal anomalies (absent axoneme ( r 2 = −0.780), missing of central singlet tubules ( r 2 = −0.611), and missing of outer doublet tubules ( r 2 = −0.738) and forward progressive sperm motility. Conclusion High levels of CP can be measured in iOAT patients, indicating that OS could underlie the aetipopathogenesis of the syndrome. OS negatively affects flagellar axonemal structure with subsequent impairment of forward progressive sperm motility. This can put an attention for antioxidants as a therapy for iOAT syndrome and further research to find how to decrease ROS production.

Rsp1 Is Indispensable for mRNA Stability during Erythroid Terminal Differentiation

Mouse models have proven invaluable for understanding erythropoiesis. Here, we describe an autosomal recessive inherited anemia in the mouse mutant hem6. Hematologic and transplantation analyses revealed a mild, congenital, hypochromic, microcytic anemia intrinsic to the hematopoietic system that is associated with a decreased red blood cell zinc protoporphyrin to heme ratio, indicative of porphyrin insufficiency. Iron uptake experiments showed that hem6 reticulocytes are defective in heme production, but not cellular iron uptake defects. Male hem6 mice are infertile due to defects in sperm structure and motility. Through positional cloning and BAC complementation, we identified the gene responsible for the hem6 anemia to be Rsp1 (RNAStabilityProtein1) on Chr.3. The anemia phenotype was reproduced by injecting zebrafish embryos with morpholinos specifically targeting the Rsp1 transcript. Multi-tissue, real-time PCR shows that Rsp1 is abundantly expressed in bone marrow and testis, which correlates with the anemia and infertility defects in the hem6 mutant. To further study the function of Rsp1, we conducted yeast two hybrid analysis, employing a human bone marrow library. We found that Rsp1 interacts with the poly (A)-binding protein cytoplasmic 1(PABPC1) and, using microarray analysis, showed that many abundant erythroid specific transcripts, including Hbb-b1, Hba-a1, Alas2, and Mitoferrin1, were significantly down regulated in hem6 reticulocytes. Since PABPC1 is responsible for stabilizing the Hba-a1 transcript [Wang et al. ( 1999) Mol Cell Biol 19:4552–60 and Wang and Kiledjian ( 2000) Mol Cell Biol 20:6334–41], we hypothesized that the relative deficiency in erythroid-specific mRNAs in hem6 reticulocytes might be due to decreased mRNA stability. Indeed, serial microarray analysis of reticulocytes aged in vitro showed that numerous, abundantly expressed erythroid-specific transcripts decayed at faster rates in hem6 reticulocytes compared to control reticulocytes. Furthermore, these mRNAs also have progressively shorter poly (A) tails, suggesting a mechanism for the increased rate of decay. In sum, the evidence supports the conclusion that Rsp1 is a regulator of mRNA stability and gene expression during the terminal differentiation of erythroid cells.

A Mutation on Mouse Chromosome 15 Causes Sperm Motility Defects and Male Infertility.

A Mutation on Mouse Chromosome 15 Causes Sperm Motility Defects and Male Infertility.

Regulation and Function of the Rhox5 Homeobox Gene

Recently, a large cluster of homeobox genes was discovered on the X chromosome that is expressed in reproductive tissues after birth. It is postulated that these reproductive homeobox genes on the X chromosome (Rhox) encode transcription factors that regulate gametogenesis. In support of this, male mice lacking the founding member of this gene cluster, Rhox5, are subfertile, exhibiting increased germ-cell apoptosis and a defect in sperm motility. To identify RHOX5 targets, microarray analyses were used to identify genes differently expressed in postnatal testes from Rhox5-null and control littermates. Highly overrepresented were genes that encode proteins involved in cellular metabolism. Several lines of evidence indicated that one of these, insulin II, is a direct target of RHOX5. Microarray analysis was also used to identify genes differentially expressed in response to physiological levels of Rhox5 in a Sertoli-cell line. Among the few genes identified, the netrin-1 receptor UNC5c, a proapoptotic molecule that is inhibited by RHOX5, was also regulated in vivo, and is thus a candidate to be downstream of RHOX5 in a prosurvival germ-cell pathway. To understand the means by which Rhox5 expression is restricted to Sertoli nurse cells in the testis, a variety of molecular approaches were used in both Sertoli-cell lines and mice. This analysis revealed that both positive and negative cis elements collaborate to confer Sertoli cell-specific gene expression. Acting on the positive cis elements are androgen receptor and GATA transcription factors. Collectively, the results of this study provide an initial glimpse into the regulatory networks that control spermatogenesis.

16 Sperm motility defects in chronic pain and fatigue syndromes

16 Sperm motility defects in chronic pain and fatigue syndromes

JAM-A is present in mammalian spermatozoa where it is essential for normal motility

Junctional adhesion molecules (JAMs) that are expressed in endothelial and epithelial cells and function in tight junction assembly, also perform important roles in testis where the closely-related JAM-A, JAM-B, and JAM-C are found. Disruption of murine Jam-B and Jam-C has varying effects on sperm development and function; however, deletion of Jam-A has not yet been studied. Here we show for the first time that in addition to expression in the Sertoli–Sertoli tight junctions in the seminiferous tubules, the ∼ 32 kDa murine JAM-A is present in elongated spermatids and in the plasma membrane of the head and flagellum of sperm. Deletion of Jam-A, using the gene trap technology, results in flagellar defects at the ultrastructural level. In Jam-A-deficient mice, which have reduced litter size, both progressive and hyperactived motility are significantly affected ( P < 0.0001) before and, more severely, after capacitation. The findings show that JAM-A is involved in sperm tail formation and is essential for normal motility, which may occur via its signal transduction and protein phosphorylation properties. Detection of JAM-A in human sperm proteins indicates that its role may be conserved in sperm motility and that JAM-A may be a candidate gene for the analysis of idiopathic sperm motility defects resulting in male subfertility in the human population.

The Macaque Sperm Actin Cytoskeleton Reorganizes in Response to Osmotic Stress and Contributes to Morphological Defects and Decreased Motility1

Sperm undergo extreme variations in temperature and osmolality during cryopreservation, resulting in cell damage that includes plasma membrane defects, changes in cell volume, decreased motility, and flagellar defects. However, the fundamental biologic mechanisms underlying these events are poorly understood. We investigated the effects of osmotic stress and cytochalasins b (CB) and d (CD), naturally occurring toxins that disrupt actin organization, on the actin cytoskeleton and motility of Rhesus macaque sperm (Macaca mulatta). Sperm were diluted in media of low, medium, or high osmolality, or medium-osmolality media containing CB or CD, were stained with phalloidin-fluorescein isothiocyanate, and were processed for microscopy. The majority of sperm incubated in medium-osmolality media exhibited postacrosomal stain, whereas the minority displayed banding patterns of F-actin stain in the head. High-osmolality media, as well as CB and CD incubation, resulted in reorganization of F-actin into bands of stain in the majority of sperm heads. Cytochalasin b treatment also resulted in curled and looped tails, a phenomenon of hyposmotic stress, and CB and CD caused significant, dose-dependent decreases in motility determined by computer-assisted sperm assessment. Rho A cell populations were determined using flow cytometry, and immunocytochemistry analysis demonstrated that Rho A localization was altered after osmotic stress. Together, our results support a mechanism in which reorganization of the actin cytoskeleton induced by osmotic stress and potentially mediated by a Rho A signaling pathway contributes to sublethal sperm flagellar and motility defects.