The identification of human sperm hyperactivated motility has potential importance in sperm function tests, as well as in quality control assays and in reproductive toxicology investigations. However, relatively little is known about this phenomenon and the variety of definitions used for hyperactivation has led to a great deal of confusion as to its occurrence and physiological relevance. This presentation is a critical review of a number of aspects of hyperactivated motility, including its identification and potential role(s) in mammalian fertilization. The initial sections of the review consider the mechanisms involved in the development and maintenance of mammalian sperm motility, and the structural and functional changes in spermatozoa which occur during transport through the female reproductive tract. The methods available for the quantification of aspects of sperm movement are also discussed, with an historical overview of sperm movement analysis.

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A critical review of the physiological importance and analysis of sperm movement in mammals

Computerized semen analysis: objective measurement of semen characteristics is biased by subjective parameter setting.

The authors examined the value of sperm creatine-kinase (CK) activity parameters to predict sperm fertilizing potential of oligospermic men. Two patient groups from our intrauterine insemination program were studied: fertile oligospermic (32 men/46 specimens) and infertile oligospermic (19 men/82 specimens). In the initial specimens, the CK activities were (mean + SEM IU CK/10(8) sperm): 0.53 +/- 0.09 and 1.17 +/- 0.19 (P less than 0.001). The corresponding values in the swim-up fractions were 0.32 +/- 0.06 and 0.67 +/- 0.08 (P less than 0.001). In a subset of samples by fertile (N = 33) and infertile (N = 66) oligospermic men who had close to identical sperm concentrations (11.9 +/- 0.9 vs. 11.9 +/- 0.5 million sperm/ml) and motility values (23.7 +/- 1.7 vs. 23.0 +/- 1.3%), the CK activities were significantly lower in the fertile group in both the initial (P = 0.02) and in the swim-up (P = 0.002) samples. A logistic regression analysis of all 160 study samples (including 21 normal men/32 samples) further demonstrated that CK activities were predictive of fertilizing potential, whereas sperm concentrations of the samples provided no additional contribution. Sperm CK and similar biochemical markers will facilitate selection of men for various approaches in assisted reproduction.

Sperm creatine kinase activity in fertile and infertile oligospermic men.

Tracks of 30 progressively motile washed human spermatozoa were plotted manually from 200-Hz frame rate video recordings. Tracks at 100, 66.7, 50, 40, 33.3, 25, 20, 10, and 5 Hz were then constructed using every 2nd, 3rd, 4th, 5th, 6th, 8th, 10th, 20th, or 40th point. The 200-Hz tracks were analyzed by traditional manual methods, and all ten sets of tracks analyzed using a computer-assisted method ("Videomot," developed originally to analyze 30-Hz tracks) to eliminate observer bias. Progression velocity (VSL) remained constant under all analysis conditions. Average path velocity (VAP) also remained essentially constant, although Videomot was less reliable at high frame rates due to problems in determining the average path. Curvilinear velocity (VCL) was very frame rate dependent (the 25-Hz mean value was only 56.5% of that at 200 Hz), and Videomot was more accurate than manual analysis at 200 Hz. Values of the amplitude of lateral head displacement (ALH) were acceptable at most frame rates. At less than 66.7 Hz the inclusion of spurious curvilinear track deviations caused lower mean ALH values, and at 5 Hz ALH could not be measured since the track was essentially the average path. Beat/cross frequency (BCF) was also frame rate dependent; at high rates there was the same problem as with ALH measurements, while at less than or equal to 25 Hz the maximum BCF was restricted by the frame rate. We conclude that human sperm movement characteristics can be measured at frame rates ca. 30 Hz but only if the constraints affecting VCL and BCF values are understood and accepted. Finally, less than 10 Hz can only give values for VSL and, perhaps, VAP.

Influence of image sampling frequency on the perceived movement characteristics of progressively motile human spermatozoa.

One hundred fourteen semen samples from Chinese males were analyzed for routine semen parameters including the semen volume, sperm count, percentage motility, and percentage normal morphology. Of these 114 samples, 54 also had movement characteristics of seminal and swim-up sperm evaluated by the computer image analyzer system (Cellsoft; Cryo Resources Co., New York). All semen samples were subjected to the swim-up procedure to harvest the motile sperm before inseminations of human oocytes. Fertilization was considered to have occurred when at least one oocyte was observed with two or more pronuclei. Semen samples were classified as infertile (0% fertilization rate;N=32) or fertile (>0% fertilization rate;N=82) before statistical analyses. There was a significant difference (P<0.005) in percentage normal morphology of seminal sperm between the fertile (mean±SE; 67.3±1.2%) and the infertile (59.3±2.2%) samples. The percentage normal morphology of seminal sperm correlated (r=0.3049;P<0.002) with the fertilization rate and this parameter was selected by the multivariate stepwise discriminant analysis as the discriminator capable of predicting the fertilization rate with 57.9% accuracy. Statistical analyses of samples where sperm movement was also evaluated demonstrated that there was significant differences (P<0.01) between the fertile (N=38) and the infertile (N=16) samples in percentage normal morphology of seminal sperm (67.8±1.8% vs 56.2±2.6%) and curvilinear velocity of swim-up sperm (89.2±3.5 vs 68.2±7.2 μm/sec). The fertilization rates correlated with the percentage normal morphology of seminal sperm (r=0.3868;P<0.005) and velocity of swim-up sperm (r=0.3842;P<0.005). Multivariate stepwise discriminant analysis demonstrated that these two sperm parameters in combination were capable of predicting the fertilization rate with 74.1% accuracy. Our results indicate that seminal sperm morphology, coupled with computerassisted image analysis of movement characteristics of swim-up sperm, can help to predict the outcome of in vitro fertilization of human oocytes.

Predictive value of sperm morphology and movement characteristics in the outcome of in vitro fertilization of human oocytes.

Use of a microcomputer with the multiple exposure photography (MEP) method for rapid analysis of semen characteristics is described. The computer is programmed to calculate information fed from a digitizing tablet while scanning projected images of photographed sperm. In this way sperm concentration, percentage of motility, average velocity and frequency distribution of sperm velocity, as well as percentage of abnormal forms are immediately obtained. The method was found to be as accurate, less prone to error, and significantly less time consuming than the previous manual measuring method. It can greatly facilitate routine semen analysis in laboratories using the MEP method.

Rapid Microcomputer-Based Analysis of Semen Characteristics from Photographs Taken by the MEP Method

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Rapid Microcomputer-Based Analysis of Semen Characteristics from Photographs Taken by the MEP Method