Effects of  -chlorohydrin on the metabolism of testicular and epididymal spermatozoa of rams

Abstract

Spermatozoa were collected from the rete testis and vas deferens of conscious rams. The endogenous oxygen uptake of the spermatozoa was unaffected by alpha-chlorohydrin added in vitro, although this compound abolished the stimulation of oxygen uptake caused by the addition of glycerol. The metabolism of [14C]glycerol by testicular and epididymal spermatozoa was markedly reduced by alpha-chlorohydrin, CO2 production and lactate accumulation being almost totally inhibited. These effects were dependent upon a period of preincubation of the spermatozoa with alpha-chlorohydrin alone, since the presence of glycerol protected the spermatozoa from its action. Longer exposure and a higher concentration of alpha-chlorohydrin were needed with testicular than with epididymal spermatozoa to achieve a maximal effect. The metabolism of [14C]glucose by both sperm types was also inhibited by alpha-chlorohyrin. Spermatozoa of the ram are therefore susceptible to the action of alpha-chlorohydrin throughout the epididymis, although more mature spermatozoa are more affected. It is suggested that alpha-chlorohydrin is converted to an intermediate which is the agent responsible for the inhibition of glycolysis in spermatozoa.

A reversible contraceptive action of some 6-chloro-6-deoxy sugars in the male rat

“…Spermatozoa were expelled from the cauda epididymidis by retrograde flushing (Ford et al, 1977) and pooled within each group. They were dispersed in phosphatebuffered saline (100 mM-NaCl, 3 mM-KCl, 6-5 mM-Na2HP03,1-5 mM-KH2P04, 0-8 mM-CaCl2 and 0-5 mM-MgCl2) (PBS) (Edwards et al, 1976) containing 90 µg penicillin G and 120 µg streptomycin sulphate/ml. Duplicate aliquots (3 ml; 0-74-0-84 IO8 spermatozoa) were incubated with 6 µ (3 uCi) D-[U-14C]glucose (Radiochemical Centre, Amersham) for 2 h at 34°C in stoppered 10 ml conical flasks in a shaking water bath.…”

“…The antifertility action of low doses of -chlorohydrin (3-chloropropan-l,2-diol) on the male (Ericsson & Youngdale, 1970) is associated with a marked reduction in the ability of spermatozoa from treated animals to oxidize glucose (Mohri, Suter, Brown-Woodman, White & Ridley, 1975;Edwards, Dacheux & Waites, 1976;Ford, Harrison & Waites, 1977) and it is possible that this may be the primary cause of the antifertility action. With the glycolytic effect in mind, we have examined the action of some chlorinated sugars on the fertility of male rats.…”

Effects of osmolality, bicarbonate and buffer on the metabolism and motility of testicular, epididymal and ejaculated spermatozoa of boars

“…The extratesticular rete was cannulated as described previously for the ram (Voglmayr et al, 1966;Voglmayr, Scott, Setchell & Waites, 1967). The rete testis fluid was drawn caudally into a vessel, cooled to 4°C as described for the ram (Edwards, Dacheux & Waites, 1976). The boars were kept in a normal pen and the catheters remained patent for up to 8 days, with a fluid flow rate of about 2-5-3 ml/h and 150 IO6 spermatozoa/ml.…”

The interaction of -chlorohydrin with glycerol kinase

“…Sufficient evidence has now accumulated to suggest that a-chlorohydrin is not itself the active compound causing disruption of sperm metabolism. For instance, a period of preincubation of ram spermatozoa with 0-1-1 mM-a-chlorohydrin is required before an inhibition of glycerol metabolism is noted (Edwards, Dacheux & Waites, 1976). Furthermore, the activity of a number of glycolytic enzymes is substantially reduced when measured in ram spermatozoa which have been incubated with 0-03-300 mM-a-chlorohydrin before extraction, yet these same enzymes are not inhibited by the direct addition of a-chlorohydrin to extracts of unincubated spermatozoa (Mohri, Suter, Brown-Woodman, White & Ridley, 1975; Brown-Woodman, Mohri, Mohri, Suter & White, 1978).…”

“…In fact, synthetic a-chlorohydrin phosphate has been shown to be an inhibitor of both sperm glyceraldehyde phosphate dehydrogenase and trióse phosphate isomerase (Mohri et al, 1975;Mashford & Jones, 1978). Moreover, high concentrations of glycerol can overcome the inhibitory effect of a-chlorohydrin on ram spermatozoa (Edwards et al, 1976;Brown-Woodman et al, 1978), suggesting competition between the two compounds for the same pathway. Such competition is supported by the report that a-chlorohydrin is a competitive inhibitor of glycerol kinase prepared from Escherichia coli (Thorner, 1972; see Brooks, Hamilton & Mallek, 1974 (Blackshaw, 1954) and the spermatozoa were separated from seminal plasma by the procedure of Harrison (1976).…”