The metabolism of ejaculated spermatozoa from the fowl

Abstract

It has been pointed out by a number of investigators, notably Lardy and Phillips ('43) and MacLeod ('41), that studies on the metabolism of ejaculated spermatozoa must be carried out using cells removed from, and washed free of the seminal fluid which normally bathes them. Except for the measurement of endogenous oxygen consumption by Lardy and Phillips ('43), and a more recent report by van Tienhoven ('60), work on cock sperm metabolism has involved the use of whole semen. Bogdonoff and Shaffner ('54) reported that the optimum pH for respiration, as measured by methylene blue reduction, of chicken semen was near 7. Zayat and van Tienhoven ('59) presented ZO?. values for cock semen before and after storage for 24 hours at 5°C in three different diluents. Kosin ('44) reported QoZ, and Qw" values, and succinate stimulated respiration of cock semen before and after X-irradiation. Later the oxidative and glycolytic activity of turkey semen was investigated (Kosin, '58), and it was suggested that quantitative aspects of oxidative activity in semen from this animal were similar to those of chicken sperm. Almost nothing is known about the metabolic characteristics of the cock spermatozoan per se. Certainly, the biochemical data obtained with these cells suspended in such a complex and undefined medium as seminal plasma are difficult to interpret, and differ considerably, as will be indicated in the present report, when compared with studies on washed cells.The present investigation is concerned with the respiration and aerobic and anaerobic glycolysis in fowl spermatozoa. In addition, data is presented suggesting breed differences in metabolism which may be of genetic significance. METHODSCock semen was collecteda by the massage technique of Burrows and Quinn ('37). For most experiments ejaculates from several birds were pooled before use in order to avoid individual variation inherent in the semen from any given animal. The cells were concentrated by gentle centrifugation (700 X G for 15 minutes) and washed two times in a calcium-free Krebs Ringer's phosphate buffered at pH 7.1 (Umbreit et al., '51).Oxygen consumption of the sperm suspension (3.0 ml final volume) was measured at 37°C in air, using the conventional Warburg apparatus according to the usual procedures (Umbreit et al., '51). The cells were placed in the sidearm of the flask and tipped into the main compartment after a 10-1 5 minute equilibration period.Glucose utilization and lactic acid production were used as criteria of glycolytic activity and were determined in Ba(OH)2-ZnSOd filtrates of the incubates. Anaerobiosis was attained by gassing the flasks for 10 minutes with commercially prepared specially purified (99.99% ) nitrogen, while shaking in the constant temperature bath at 37"C, after which, at zero time, the cells were tipped into the main compartments of the flasks.Glucose was estimated by the NelsonSomogyi (Somogyi, ' 5 2 ) colorimetric procedure. Lactic acid was determined by the method of Barker and Summerson ('41).

The Distribution of Some Hydrolytic Enzymes in the Semen of the Domestic Fowl, Gallus Domesticus

“…This recalls the results of previous studies on the fowl and turkey where between-breed, between-family and between-bird differences were found in other metabolic features of spermatozoa (Goldberg & Norman, 1961;Mclndoe & Lake, 1969Buckland, 1971). Between-breed differences have also been demonstrated in the survival following storage of spermatozoa in vitro (Wilcox, Shaffner & Wilson, 1961;Lake, 1972) and, apart from this, differences in fertility pheno¬ mena are known to exist between types of domestic birds; it will be of interest to discover whether or not a direct relationship exists between the abovementioned basic metabolic features and functional aspects of sperm activity.…”

Effects of High Pressure Gases on the Motility and Fertilizing Capacity of Avian Spermatozoa Stored in Vitro

“…It was assumed that oxygen was beneficial in the support of the endogenous metabolism of spermatozoa (Goldberg & Norman, 1961). This theory is further supported by the significantly higher fertilizing capacity of spermatozoa stored under air at 21 atm since the latter would produce a higher concentration of oxygen over the semen surface, thereby promoting the oxidative processes associated with sperm metabolism.…”

Reduced Glucose Transport in Sperm from Roosters (Gallus domesticus) with Heritable Subfertility1

“…Fowl sperm can produce ATP under anaerobic conditions to maintain motility and fertilizing ability [28,29]. Even though there is not a sperm specific-LDH in fowl sperm [30][31][32], LDH is critical for ATP production by anaerobic glycolysis [25].…”