Studies of the Distribution and Source of Enzymes in Mammalian Semen

Murdoch, R. N.; White, IG

doi:10.1071/bi9680483

Cited by 36 publications

(30 citation statements)

References 7 publications

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“…Glucose and lactate were estimated in protein-free extracts of the spermatozoal suspension at the beginning and end of all metabolic studies by procedures previously described (Murdoch and White 1968). The 14C02 produced by the spermatozoa from D-[l_14C]-and D-[6-14C]glucose was assayed as Ba14C03 by the procedure of Annison and White (1961).…”

Section: (D) Analytical Methodsmentioning

confidence: 99%

“…Absence of Bicarbonate Hamner and Williams (1964) and Murdoch and White (1968) have shown that bicarbonate is the factor in tubal and uterine fluids stimulating spermatozoal metabolism. It was of interest, therefore, to study the metabolism of spermatozoa after storage in the presence of bicarbonate to see if it would produce a response similar to that obtained with oviduct and uterine fluids in experiment 1.…”

Section: (C) Experiments 3: Metabolism Of Labelled Glucose After Storamentioning

confidence: 99%

“…Oviduct and uterine fluids were collected from oestrous does as described in Part I of this series (Murdoch and White 1968) (Umbreit, Burris, and Stauffer 1959) to give the desired concentration and radioactivity_ The diluent contained streptomycin and penicillin (0· 5 mg/ml of each). The radioactivity of the labelled substrates was checked by counting in a Nuclear Ohicago liquid scintillation spectrophotometer.…”

Section: (A) Genital Tract Fluidsmentioning

confidence: 99%

“…Since both tubal and uterine fluid contain bicarbonate, which stimulates spermatozoal respiration and glycolysis Williams 1963, 1964;Murdoch and White 1968), it may in some way be related to the metabolie changes occurring in spermatozoa after incubation in utero. If, however, bicarbonate is involved, it must precondition the spermatozoa to such metabolic changes as they were washed free of luminal secretions after removal from the uterus and incubated in Warburg flasks containing potassium hydroxide to absorb carbon dioxide (Hamner and Williams 1963;Mounib and Chang 1964;Murdoch and White 1967).…”

Section: Introduotionmentioning

confidence: 99%

“…From this work, it appears that capacitated rabbit spermatozoa may have greater respiratory and glycolytic activity than freshly ejaculated spermatozoa, and may oxidize C-l of the glucose molecule in preference to C-6, indicating Sci., 1968, 21, 961-72. a shift towards pentose shunt activity. Freshly ejaculated spermatozoa oxidize C-l and C-6 of glucose at a similar rate suggesting that the sugar is metabolized predominantly via the Embden-Meyerhof glycolytic pathway.Since both tubal and uterine fluid contain bicarbonate, which stimulates spermatozoal respiration and glycolysis Williams 1963, 1964;Murdoch and White 1968), it may in some way be related to the metabolie changes occurring in spermatozoa after incubation in utero. If, however, bicarbonate is involved, it must precondition the spermatozoa to such metabolic changes as they were washed free of luminal secretions after removal from the uterus and incubated in Warburg flasks containing potassium hydroxide to absorb carbon dioxide (Hamner and Williams 1963;Mounib and Chang 1964;Murdoch and White 1967).…”

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confidence: 99%

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The Influence of the Female Genital Tract on the Metabolism of Rabbit Spermatozoa II. Effect of Storage With Glucose Lactate, Bicarbonate , and Female Genital Tract Fluids

Rn¹,

White²

1968

Aust. Jnl. Of Bio. Sci.

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SummaryIn an attempt to isolate the factors responsible for altering the metabolism of spermatozoa incubated in utero, washed rabbit spermatozoa were stored in vitro for 6 hr at 37°C with either glucose, lactate, bicarbonate, or female genital tract fluids, and their ability to metabolize [1_14C]glucose and [6-14C]glucose was then compared with that of fresh spermatozoa.Spermatozoa suffered a decline in oxygen uptake and glycolysis after storage for 6 hr and the yield of 14C02 from [6-14C]glucose was less than that from [l_14C} glucose. Adding glucose or lactate to the medium lessened these senescence changes and the spermatozoa retained their ability to oxidize C-1 and C-6 of glucose at about the same rate. Low concentrations of uterine fluid, but not bicarbonate, had a similar effect. When higher concentrations of uterine fluid were included in the storage diluent with glucose, subsequent glycolytic activity, but not oxygen uptake, was depressed.No addition to the incubation medium produced the changes characteristic of spermatozoa recovered from the uterus (i.e. increase in glycolysis, oxygen uptake, and oxidation of C-1 of glucose above the fresh controls).It is concluded that these changes cannot be accounted for solely by the beneficial effects of genital tract fluids, glucose, or lactate, and some other unknown property of the uterus must also be involved. The evidence suggests that it is not bicarbonate. INTRODUOTIONAlthough over a decade has passed since the discovery that spermatozoa must spend 2-6 hr in the female reproductive tract before fertilization can occur (Austin 1951(Austin , 1952 Chang 1951;Noyes 1953), no biochemical changes in the spermatozoa have been unequivocally related to this process of capacitation. Recently, however, attempts have been made to approach the problem by comparing the metabolism of freshly ejaculated rabbit spermatozoa with that of spermatozoa after incubation in the uterus for 6 hr (Hamner and Williams 1963;Mounib and Chang 1964;Murdoch and White 1967). From this work, it appears that capacitated rabbit spermatozoa may have greater respiratory and glycolytic activity than freshly ejaculated spermatozoa, and may oxidize C-l of the glucose molecule in preference to C-6, indicating Sci., 1968, 21, 961-72. a shift towards pentose shunt activity. Freshly ejaculated spermatozoa oxidize C-l and C-6 of glucose at a similar rate suggesting that the sugar is metabolized predominantly via the Embden-Meyerhof glycolytic pathway.Since both tubal and uterine fluid contain bicarbonate, which stimulates spermatozoal respiration and glycolysis Williams 1963, 1964;Murdoch and White 1968), it may in some way be related to the metabolie changes occurring in spermatozoa after incubation in utero. If, however, bicarbonate is involved, it must precondition the spermatozoa to such metabolic changes as they were washed free of luminal secretions after removal from the uterus and incubated in Warburg flasks containing potassium hydroxide to absorb carbon dioxide (Hamner and Williams 1963;M...

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Section: (D) Analytical Methodsmentioning

confidence: 99%

Section: (C) Experiments 3: Metabolism Of Labelled Glucose After Storamentioning

confidence: 99%

Section: (A) Genital Tract Fluidsmentioning

confidence: 99%

Section: Introduotionmentioning

confidence: 99%

mentioning

confidence: 99%

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The Influence of the Female Genital Tract on the Metabolism of Rabbit Spermatozoa II. Effect of Storage With Glucose Lactate, Bicarbonate , and Female Genital Tract Fluids

Rn¹,

White²

1968

Aust. Jnl. Of Bio. Sci.

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Cell surface changes in the proteins of rabbit spermatozoa during epididymal passage

et al. 1979

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Differences in the exposure of spermatozoa surface components during epididymal passage have been examined using lactoperoxidase‐catalyzed 125I‐iodination or labeling with 125I‐diazodiiodosulfanilic acid. Labeled surface proteins obtained from caput and cauda epididymides were solubilized in detergent, separated by sodium dodecylsulfate polyacrylamide slab gel electrophoresis, and identified by radiography. Densitometer scans of autoradiograms revealed increased amounts or exposures of surface proteins of ∼35,000, ∼39,000, ∼50,000, and ∼78,000 molecular weight on the cauda epididymal spermatozoa.

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Action of phosphatidylcholine in protecting ram sperm from cold shock

1986

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Rapid cooling (cold shocking) of washed ejaculated ram sperm to 0°C irreversibly reduced motility, tail beat frequency, and respiration and increased the uptake of 45Ca2+. The plasma membranes were removed from the sperm head, and the acrosomes were detached from the nuclei. The plasma membranes of the middle piece were removed, and the mitochondria contained pale and expanded cristae, similar in appearance to ATP‐deprived mitochondria in the “condensed” configuration. The presence of 2.0 mg/ml phosphatidylcholine (lecithin) in the medium prevented ultrastructural damage on cold shock, and the motility, tail beat frequency, respiratory rate, and calcium uptake were maintained at levels similar to washed sperm. As the “protective” effect of phosphatidylcholine against cold shock was maintained to a certain extent after rewashing and centrifuging the sperm prior to cold shock, the interaction of phosphatidylcholine with ram sperm membranes may be fairly “tight” and not easily disrupted.

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Studies of the Distribution and Source of Enzymes in Mammalian Semen

Cited by 36 publications

References 7 publications

The Influence of the Female Genital Tract on the Metabolism of Rabbit Spermatozoa II. Effect of Storage With Glucose Lactate, Bicarbonate , and Female Genital Tract Fluids

The Influence of the Female Genital Tract on the Metabolism of Rabbit Spermatozoa II. Effect of Storage With Glucose Lactate, Bicarbonate , and Female Genital Tract Fluids

Cell surface changes in the proteins of rabbit spermatozoa during epididymal passage

Action of phosphatidylcholine in protecting ram sperm from cold shock

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