The locus of flicker adaptation in the migraine visual system: a dichoptic study.

BACKGROUND: Flickering light has been shown to sensitize the migraine visual system at high stimulus contrast while elevating thresholds at low contrast. The present study employs a dichoptic psychophysical paradigm to ask whether the abnormal adaptation to flicker in migraine occurs before or after the binocular combination of inputs from the two eyes in the visual cortex. METHODS: Following adaptation to high contrast flicker presented to one eye only, flicker contrast increment thresholds were measured in each eye separately using dichoptic viewing. RESULTS: Modest interocular transfer of adaptation was seen in both migraine and control groups at low contrast. Sensitization at high contrast in migraine relative to control participants was seen in the adapted eye only, and an unanticipated threshold elevation occurred in the non-adapted eye. Migraineurs also showed significantly lower aversion thresholds to full field flicker than control participants, but aversion scores and increment thresholds were not correlated. CONCLUSIONS: The results are simulated with a three-stage neural model of adaptation that points to strong adaptation at monocular sites prior to binocular combination, and weaker adaptation at the level of cortical binocular neurons. The sensitization at high contrast in migraine is proposed to result from stronger adaptation of inhibitory neurons, which act as a monocular normalization pool.

Detection and discrimination of flicker contrast in migraine.

AIMS: Flickering light is strongly aversive to many individuals with migraine. This study was designed to evaluate other abnormalities in the processing of temporally modulating visual stimulation. METHODS: We measured psychophysical thresholds for detection of a flickering target and for the discrimination of suprathreshold flicker contrasts (increment thresholds) in 14 migraineurs and 14 healthy controls with and without prior adaptation to high-contrast flicker. Visual discomfort (aversion) thresholds were also assessed. RESULTS: In the baseline (no adaptation) conditions, detection and discrimination thresholds did not differ significantly between groups. Following adaptation, flicker detection thresholds were elevated equivalently in both groups; however, discrimination thresholds were more strongly affected in migraineurs than in controls, showing greater elevation at moderate contrasts and greater threshold reduction (sensitisation) at high contrast (70%). Migraineurs also had significantly elevated discomfort scores, and these were significantly correlated with number of years with migraine. DISCUSSION: We conclude that visual flicker not only causes discomfort but also exerts measurable effects on contrast processing in the visual pathways in migraine. The findings are discussed in the context of the existing literature on habituation, adaptation and contrast-gain control.

Discrimination and identification of periodic motion trajectories.

Humans are extremely sensitive to radial deformations of static circular contours (F. Wilkinson, H. R. Wilson, & C. Habak, 1998). Here, we investigate detection and identification of periodic motion trajectories defined by these radial frequency (RF) patterns over a range of radial frequencies of 2-5 cycles. We showed that the average detection thresholds for RF trajectories range from 1 to 4 min of arc and performance improves as a power-law function of radial frequency. RF trajectories are also detected for a range of speeds. We also showed that spatiotemporal global processing is involved in trajectory detection, as improvement in detection performance with increasing radial deformation displayed cannot be accounted for by local probability summation. Finally, identification of RF trajectories is possible over this RF range. Overall thresholds are about 6 times higher than previously reported for static stimuli. These novel stimuli should be a useful tool to investigate motion trajectory learning and discrimination in humans and other primates.

Protein composition of human epididymosomes collected during surgical vasectomy reversal: a proteomic and genomic approach.

BACKGROUND: The epididymal epithelium secretes membranous vesicles, called epididymosomes, with which a complex mixture of proteins is associated. These vesicles transfer to spermatozoa selected proteins involved in sperm maturation. Epididymosomes in the human excurrent duct have been described, but their protein composition and possible functions are unknown. METHODS AND RESULTS: Epididymosomes were collected during vasovasostomy procedures, purified and submitted to liquid chromatography with hybrid quadrupole time-of-flight mass spectrometry. From all the mass spectra generated, 1022 peptides allowed the identification of 146 different proteins. Identification of some of these proteins was confirmed by western blots. Furthermore, western blot showed that the protein composition of epididymosomes differed from that characterizing prostasomes; membranous vesicles secreted by the prostate. Organization of the epididymosomes proteome according to common functional features suggests that epididymosomes have multiple functions. In order to understand the origin of epididymosomes collected distally, microarray databases of caput, corpus and cauda epididymidis were analysed to determine where along the excurrent duct the encoded proteins associated to epididymosomes are synthesised. Results suggest that some proteins synthesized in the caput and corpus epididymidis are associated with epididymosomes collected distally. CONCLUSIONS: Epididymosomes thus transit along the excurrent duct, and vesicles collected distally represent a mixed population.

Polyol pathway in human epididymis and semen.

Two enzymes are involved in the polyol pathway: an aldose reductase that reduces glucose in sorbitol followed by its oxidation in fructose by sorbitol dehydrogenase. It has been previously shown that both enzymes are presented in the bovine epididymis, where they are associated with membranous vesicles called epididymosomes. Based on the distribution of these enzymes, it has been hypothesized that the polyol pathway can modulate sperm motility during the epididymal transit. In the present study, polyol pathway was investigated in semen and along the epididymis in humans in order to determine if sperm maturation can be associated with this sugar pathway. Western blot analysis shows that both aldose reductase and sorbitol dehydrogenase are associated with ejaculated spermatozoa and prostasomes in humans. These enzymes are also associated with epididymosomes collected during surgical vasectomy reversal. Western blot, Northern blot, and reverse transcription-polymerase chain reaction analysis show that aldose reductase and sorbitol dehydrogenase are expressed at the transcriptional and translational levels along the human epididymis. Unlike what occurs in the bovine model, distribution of these enzymes is rather uniform along the human excurrent duct. Immunohistological studies together with Western blot analysis performed on epididymosomes preparations indicate that the polyol pathway enzymes are secreted by the epididymal epithelium. These results indicate that the polyol pathway plays a role in human sperm physiology.

Gene expression in the epididymis of normal and vasectomized men: what can we learn about human sperm maturation?

Anatomically, the human epididymis is unusual when compared with the excurrent duct of other eutherian mammals. Furthermore, clinical observations suggest that it may not be as important for sperm maturation as is the case for laboratory animals. In contrast, hierarchical clustering of microarray data of epididymides from normal men revealed 2274 modulated qualifiers between the epididymal segments, 1184, 713, and 269 of them being highly expressed in the caput, corpus, and cauda, respectively. The organization of qualifiers according to their similarities by gene ontology indicated that caput transcripts are dedicated to cell-cell adhesion, whereas the corpus is characterized by genes involved in response to other organisms (ie, defense mechanisms) and the cauda transcriptome is specialized in muscle contraction and establishment of localization. A region-specific gene expression pattern thus characterizes the human epididymis as in animal models. In humans, vasectomies have consequences on the epididymal transcriptome. Cluster analysis revealed that 1363 genes are expressed in both normal and vasectomized epididymides, whereas 911 and 660 of them are specifically expressed in normal and vasectomized epididymides, respectively. Three of the affected genes are particularly interesting because of their involvement in sperm biochemical remodeling during epididymal transit: dicarbonyl/l-xylulose reductase, Niemann-Pick disease, type C2, and cysteine-rich secretory protein 1. In some vasovasostomized men, these modifications in gene expression induced by vasectomy are irreversible, thus affecting the biochemical parameters, and potentially, the function of their ejaculated sperm. This may explain the discrepancies between a surgically successful vasovasostomy and fertility recovery.

Vasectomy affects cysteine-rich secretory protein expression along the human epididymis and its association with ejaculated spermatozoa following vasectomy surgical reversal.

The epididymis is essential for the acquisition of sperm fertilizing ability and forward motility. After vasectomy, the flux and composition of the epididymal fluid are modified, causing possible sequelae to the occluded excurrent duct. Some of these sequelae may not be reversible following vasovasostomy, affecting sperm physiology and their fertilizing ability. We previously demonstrated that the epididymal expression in men of a major glycoprotein secreted by the epididymis, cysteine-rich secretory protein 1 (CRISP1), and its encoding mRNA are affected by vasectomy. In this study we showed that following vasectomy, the increased level of CRISP1 is not due to a secretory defect but to its accumulation in the intraluminal compartment of the cauda epididymidis. Western blot analyses were performed to determine the amount of CRISP1 associated with spermatozoa of men who had undergone surgical vasectomy reversal. Spermatozoa of vasovasostomized men are characterized by a significant increase (P < .05) in CRISP1 levels when compared with normal donors. There was no linear correlation between CRISP1 levels and the period of time elapsed between vasectomy and vasovasostomy. CRISP1 was also present in seminal plasma of normal and vasovasostomized men, but not in vasectomized individuals. The soluble concentration of CRISP1 was significantly higher (P < .05) in seminal plasma of vasovasostomized men when compared with normal men. Knowing that one of the proposed functions of CRISP1 is to modulate sperm capacitation, we evaluated the level of tyrosine protein phosphorylation of 2 AXAP proteins of the fibrous sheath, p81 and p105. Spermatozoa of vasovasostomized men were characterized by a 50% increase of protein tyrosine phosphorylation when compared to spermatozoa of normal men (P < .05). Our results are discussed with regard to the fertilizing ability of ejaculated spermatozoa of some vasovasostomized men.

Factors predicting overall success: a review of 747 microsurgical vasovasostomies.

OBJECTIVES: Advances in surgical techniques have improved the outcome of microsurgical vasovasostomy (VV). We performed a retrospective analysis of surgical procedures to determine outcomes and predictors of VV success, to develop Kaplan-Meier Curves for predicting VV outcomes and to evaluate the use of alpha-glucosidase (AG) to predict outcomes. PATIENTS AND METHODS: We undertook a retrospective analysis of 747 modified 1-layer microsurgical VV procedures performed between 1984 and 2000. Obstructive interval, partner status, social status preoperatively and method of vasal obstruction, vasal fluid quality and sperm granuloma intraoperatively were compared with outcome results. Parameters evaluated at follow-up included semen analysis, AG concentration in ejaculate fluid and pregnancy rates. RESULTS: The overall patency rate was 86% and pregnancy rates were 33% and 53% at 1 and 2 years after primary VV, respectively. Preoperative factors associated with successful outcome and pregnancy included shorter obstructive interval and same female partner (p < 0.05). Intraoperative factors predicting success included the use of surgical clips instead of suture at vasectomy, the presence of a sperm granuloma, the presence and quality of vasal fluid, and the presence and quality of sperm in vasal fluid. Further, increased AG in the postoperative semen predicted improved patency and pregnancy outcomes. CONCLUSION: This study confirms the effectiveness of VV for vasectomized men who wish to father children. It also demonstrates that preoperative and intraoperative factors are predictive of the VV outcome. Postoperative AG is also a useful marker of patency and it appears to predict pregnancy outcome.

HE1/NPC2 status in human reproductive tract and ejaculated spermatozoa: consequence of vasectomy.

We have previously demonstrated that the amount of HE1/NPC2 mRNA and protein expressed in the human epididymis is decreased under vasectomy. In this study, western blot analyses showed that many vasovasostomized men are characterized by high HE1/NPC2 levels in spermatozoa when compared with fertile donors. HE1/NPC2 association with sperm from vasovasostomized men was not related to low motility per se as spermatozoa from asthenospermic men have HE1/NPC2 levels similar to those in normal fertile semen samples. Spermatozoa from vasovasostomized men with high amount of HE1/NPC2 are characterized by higher concentration of cholesterol and more lipid raft domains. HE1/NPC2 is secreted in different glycoforms by different tissues of human male reproductive tract. These forms are due to variation in N-glycosylation, and only the deglycosylated form is associated with spermatozoa from some vasovasostomized men. Compared with normal men, seminal plasma of vasectomized men is characterized by a major decrease in immunodetectable HE1/NPC2 without change in the glycosylation pattern. Following surgical vasectomy reversal, seminal plasma HE1/NPC2 was found in similar amounts to the ones characterizing normal men. Considering the potential role of HE1/NPC2 in cholesterol transport during sperm maturation, unusual high levels of this protein associated with spermatozoa of vasovasostomized men may reflect epididymal sequelae occurring when the vas deferens is obstructed.

Expression of heat shock protein 70 in normal and cryptorchid human excurrent duct.

The Hsp70 heat-shock proteins are molecular chaperones that assist other proteins in their folding, transport and assembly into complexes. Most of these proteins are either constitutively expressed or induced by heat shock and other stresses. Heat shock proteins are required for spermatogenesis, and also protect cells from environmental hazards such as heat, radiation, and chemicals. The abdominal position of the cryptorchid testis provokes a temperature elevation which is detrimental to spermatogenesis and causes infertility. The consequences of such a stress on Hsp70 expression were evaluated in normal and cryptorchid epididymides and vas deferens. Hsp70-1 transcript from reproductive organs of normal and cryptorchid men was analysed by real-time quantitative RT-PCR, meanwhile Hsp70 protein was characterized by western blot and immunohistochemical staining analysis. Hsp70-1 mRNA and protein showed equal expression in all segments of normal epididymis and ductus deferens. Hsp70 was specific to basal cells in the epididymal epithelium. For cryptorchid patients, Hsp70-1 mRNA expression in caput epididymidis was unchanged compared with controls. However, in corpus and cauda epididymides as well as in vas deferens, the expression level of Hsp70-1 transcript was higher for the cryptorchid tissues. Changes in mRNA frequency were specifically correlated with the age of the patients. By opposition to the mRNA, western blot analysis revealed that Hsp70 protein levels were not affected by the inguinal location of the epididymis. The data show that Hsp70-1 transcript and protein were constitutively expressed in human excurrent duct and that an inguinal location can stimulate the expression of Hsp70-1 mRNA along the human epididymis and ductus deferens.