The `Sperm-Rise' Test

Abstract

After centrifugation of a mobile-sperm suspension, the spermatozoa will begin to move upwards from the sediment into the supernatant. This fact constitutes the background to the following method for comparative estimation of thè amounts of motility' in different test suspensions. The method is simple, adequately objective, and recordings can be made in parallel on several different test suspensions in which the spermatozoa have been in contact with various media for an identical time.Test suspensions containing 0\m=.\5% of cells (see below) were prepared from well-mixed stock suspensions of spermatozoa pooled from the cauda epididymidis of a varying number of gonads from slaughtered bulls. When the effects of different drugs upon sperm kinetic activity were studied, the suspending medium was a Ringer solution in which the 100-ml 0\m=.\9% NaCl (0\m=.\154m) component of Mann's Ringer solution (Mann, 1964) was replaced with a 0\m=.\191 m-NaCl solution. This replacement rendered the Ringer solution approximately isosmotic with the fluids from the cauda epipidymidis, which have a freezingpoint depression of \ m=-\ 0\ m=. \ 66\ s=deg\ C (Salisbury & Cragle 1956). When the effects of different ions were investigated, the ionic composition of the 100-ml component of the Ringer solution was altered. All suspending media also contained 0\m=.\1% glucose.Experiments on spermatoza from different stock suspensions were preceded by a preliminary test, in which a 2-0-ml sample of a control suspension con¬ sisting of 0-15 ml of stock suspension +10-0 ml of medium was centrifuged at room temperature (23 to 24°G ) for 15 min at 2000 g. The medium consisted of a mixture of 0-191 M-NaCl and NaF Ringer solution in the proportion of 15:1, in which the sperm motility was immediately blocked by the fluoride (see Lindahl & Wedin, 1963). With the aid of the spermatocrit values then obtained (which usually fell within the range of 0-008 to 0-012 ml), the quantity of stock suspension was calculated which, after mixing with 100 ml of a given test medium and subsequently centrifuging in a 2-0-ml sample, would-if the spermatozoa were immotile-give a spermatocrit value of 0-010 ml. This procedure ensured that the 'sperm rises' determined for different test suspen¬ sions in different experiments could be related to the same spermatocrit value.In the calibrated spermatocrit tubes used (AB Grave, Stockholm, Sweden), the lengths of the narrow parts, which were graded in volume fractions from zero to 0-080 ml (Text- fig. 1), were almost identical (range 54-5 to 55-5 mm), i.e. a volume fraction of 0-001 ml had a height of approximately 55/80, or 0-69 mm. 427

The Permeability of Bull Spermatozoa to Water, Polyhydric Alcohols and Univalent Anions and the Effects of the Anions Upon the Kinetic Activity of Spermatozoa and Sperm Models

“…Bull spermatozoa were obtained from the cauda epididymidis of gonads from slaughtered bulls and collected in NaCl Ringer solution (see below) as in previous studies (Drevius, 1971a(Drevius, , 1972a.…”

“…The concentration of spermatozoa in different test suspensions was adjusted to 0-5% with the aid of the preliminary test described by Drevius (1971a). A sample (2-0 ml) of a given test suspension made up in isotonic, Ringer (see below) will, provided that the spermatozoa are immotile, give a spermatocrit of 0-010 ml when centrifuged for 15 min at 2000 g in tubes of the type described in Drevius (1971a).…”

“…A sample (2-0 ml) of a given test suspension made up in isotonic, Ringer (see below) will, provided that the spermatozoa are immotile, give a spermatocrit of 0-010 ml when centrifuged for 15 min at 2000 g in tubes of the type described in Drevius (1971a).…”

“…Suspending media for different experiments (A) The influence upon sperm motility of different anions was investigated by the 'sperm-rise' test (Drevius, 1971a, and below) on cells suspended in media consisting of the Ringer solution described by Mann (1964), in which the 100 ml of 0-9% (0-154 M-)NaCl component was replaced with 0-191 M-NaCl, or with 0-191 m solutions of NaN03, NaBr, Nal or NaSCN. The osmolalities of these Ringer solutions are approximately 0-353 osmol, i.e.…”

A simple method to measure drug effects on human sperm motility.

“…We have used this method to measure the effect of caffeine on sperm motility. The stimulatory effect of caffeine was first demonstrated in epididymal (Drevius, 1971) and ejaculated (Garbers, First, Sullivan & Lardy, 1971) bovine sperms. Several studies (Bunge, 1973;Haesungcharern & Chulavatnatol, 1973;Schoenfeld, Amelar & Dubin, 1973;Levin et al, 1980;Makler et al, 1980) have shown a similar response in human sperms, although one study (Dougherty, Cockett & Urry, 1976) failed to confirm it.…”