Effects of Dilution and Washing on Ram Spermatozoa Studied by the Flow Dialysis Technique

Dott, H. M.; Walton, Arthur

doi:10.1530/jrf.0.0010350

Cited by 35 publications

(33 citation statements)

References 20 publications

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“…Sperm agglutination occurs between spermatozoa with intact plasma and acrosome membranes (Yang et al 2012). Typically head-to-head agglutination takes place when sperm is exposed in vitro to: i) washing media (ram (Dott & Walton 1960)), ii) fluids from the female genital tract (bull (Lindahl 1966)), iii) divalent cations (rabbit (Bedford 1970), bull (Lindahl 1973), iv) bovine serum in combination with semen extender (bull (Senger & Saacke 1976)) and v) IVF medium (boar (Harayama et al 1998(Harayama et al , 2000a). Divalent cations (including Ca 2C , Mg 2C and Mn 2C ) activate an ATPdependent surface reaction which triggers head-to-head sperm agglutination (Harayama et al 1998, 2000a, Yang et al 2012.…”

Section: Discussionmentioning

confidence: 99%

Combined albumin and bicarbonate induces head-to-head sperm agglutination which physically prevents equine sperm–oviduct binding

Leemans¹,

Gadella²,

Stout³

et al. 2016

Reproduction

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In many species, sperm binding to oviduct epithelium is believed to be an essential step in generating a highly fertile capacitated sperm population primed for fertilization. In several mammalian species, this interaction is based on carbohydrate-lectin recognition.D-galactose has previously been characterized as a key molecule that facilitates sperm-oviduct binding in the horse. We used oviduct explant and oviduct apical plasma membrane (APM) assays to investigate the effects of various carbohydrates; glycosaminoglycans; lectins; S-S reductants; and the capacitating factors albumin, Ca 2C and HCO 3 K on sperm-oviduct binding in the horse. Carbohydratespecific lectin staining indicated that N-acetylgalactosamine, N-acetylneuraminic acid (sialic acid) and D-mannose or D-glucose were the most abundant carbohydrates on equine oviduct epithelia, whereas D-galactose moieties were not detected. However, in a competitive binding assay, sperm-oviduct binding density was not influenced by any tested carbohydrates, glycosaminoglycans, lectins or D-penicillamine, nor did the glycosaminoglycans induce sperm tail-associated protein tyrosine phosphorylation. Furthermore, N-glycosidase F (PNGase) pretreatment of oviduct explants and APM did not alter sperm-oviduct binding density. By contrast, a combination of the sperm-capacitating factors albumin and HCO 3 K severely reduced (O10-fold) equine sperm-oviduct binding density by inducing rapid head-to-head agglutination, both of which events were independent of Ca 2C and an elevated pH (7.9). Conversely, neither albumin and HCO 3 K nor any other capacitating factor could induce release of oviduct-bound sperm. In conclusion, a combination of albumin and HCO 3 K markedly induced sperm head-to-head agglutination which physically prevented stallion sperm to bind to oviduct epithelium.

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Section: Discussionmentioning

confidence: 99%

Combined albumin and bicarbonate induces head-to-head sperm agglutination which physically prevents equine sperm–oviduct binding

Leemans¹,

Gadella²,

Stout³

et al. 2016

Reproduction

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“…Dott & Walton (1960) investigated the effects of washing on spermatozoa and published data on the removal of seminal plasma constituents from sperm suspensions using a classical washing procedure. Harrison & White (1972) published results which indirectly indicated the efficiency of their washing procedure.…”

Section: Discussionmentioning

confidence: 99%

“…Some components of seminal plasma interact with the spermatozoa (e.g. the so-called 'sperm-coating antigen' (Weil, 1967)): general measurement of removal of seminal plasma proteins (Dott & Walton, 1960) (1964) reported that rabbit spermatozoa exposed to Ficoll solutions of up to 30% (w/w) were both motile and fertile, and Prakash (1974) reported that bull spermatozoa exposed to 14 % (w/v) Ficoll were very motile. It seems most likely that the loss of motility observed after washing by the dense-layer method is an expression of the 'dilution' effect (Mann, 1964) …”

Section: Discussionmentioning

confidence: 99%

A highly efficient method for washing mammalian spermatozoa

Harrison¹

1976

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“…For example, Emmens & Swyer (1948) and Blackshaw (1953) showed that sperm motility was depressed, while Mann & Lutwak-Mann (1948), Dott & Walton (1960) and Wales & Wallace (1965) observed a reduction of metabolic activity. Workers from several laboratories have measured the loss of a number of intracellular substances, including inorganic ions (Dott & White, 1964), hexokinase and glucose phosphate isomerase (Harrison & White, 1972), glyceraldehyde-3-phosphate dehydro¬ genase (Smith, Mayer & Merilan, 1957), cytochrome (Mann, 1951), plasma¬ logen, protein, hyaluronidase and lipoprotein, from the acrosome (Mann, 1964).…”

Section: Introductionmentioning

confidence: 99%

“…Workers from several laboratories have measured the loss of a number of intracellular substances, including inorganic ions (Dott & White, 1964), hexokinase and glucose phosphate isomerase (Harrison & White, 1972), glyceraldehyde-3-phosphate dehydro¬ genase (Smith, Mayer & Merilan, 1957), cytochrome (Mann, 1951), plasma¬ logen, protein, hyaluronidase and lipoprotein, from the acrosome (Mann, 1964). In addition to the reduction in metabolic activity, Dott & Walton (1960) noticed that washing caused an increase in the proportion of eosinophilic spermatozoa and an increased tendency towards agglutination. These ob¬ servations suggest that washing causes a change in the permeability of the plasma membrane associated with the removal of antiagglutinins.…”

Section: Introductionmentioning

confidence: 99%

The Effects of Washing on the Ultrastructure and Cytochemistry of Ram Spermatozoa

Jones¹,

Holt²

1974

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Summary. Washing ram spermatozoa two or four times with Krebs\x=req-\ Henseleit-Ringer solution, to achieve dilution rates of 100-or 10, 000\ x=req-\ fold, damaged the plasma membranes, acrosomes and mitochondria of some spermatozoa. The proportion of spermatozoa with broken plasma membranes over the acrosome increased with the number of washes.Washing once to achieve a dilution rate of 10-fold was sufficient to remove from spermatozoa all the substrate available for NADPH diaphorase and probably glucose-6-phosphate dehydrogenase, but not quite all the substrate for succinic dehydrogenase. A 10-fold dilution was also sufficient to remove glucose-6-phosphate dehydrogenase from a considerable proportion of spermatozoa, but even three washes to achieve a dilution rate of 1000-fold had little effect on succinic dehydrogenase and NADPH diaphorase activity in spermatozoa.The effects of washing could not be attributed to repeated centrifugation of spermatozoa during the process.

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Effects of Dilution and Washing on Ram Spermatozoa Studied by the Flow Dialysis Technique

Cited by 35 publications

References 20 publications

Combined albumin and bicarbonate induces head-to-head sperm agglutination which physically prevents equine sperm–oviduct binding

Combined albumin and bicarbonate induces head-to-head sperm agglutination which physically prevents equine sperm–oviduct binding

A highly efficient method for washing mammalian spermatozoa

The Effects of Washing on the Ultrastructure and Cytochemistry of Ram Spermatozoa

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