A modified trans‐membrane migration method for measuring the effect of drugs on sperm motility.

Raoof, N. T.; Pearson, Richard M.; Turner, Paul

doi:10.1111/j.1365-2125.1987.tb03175.x

Cited by 8 publications

(4 citation statements)

References 8 publications

(16 reference statements)

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“…This test measures the proportion of sperm moving across the 5 ~m pores of a Nucleopore membrane from an aliquot of semen to buffer solution during 2 h incubation at 37°C. Though this method appears to be simple, quantitative, and reproducible, it suffers, besides possible problems concerning exposure conditions (Raoof et al, 1987), from the disadvantage that only overall changes in sperm swimming activity and not changes in the moving pattern of individual spermatozoa can be recorded. Computer videomicrography, however, offers the possibility to detect and analyze subtle effects of chemical compounds on the swimming pattern of spermatozoa.…”

Section: Discussionmentioning

confidence: 98%

Bovine spermatozoa as anin vitro model for studies on the cytotoxicity of chemicals: effects of chlorophenols

1989

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The suitability of ejaculated bovine spermatozoa as an in vitro model for the assessment of the cytotoxic potential of chemicals was evaluated using several endpoints: swimming activity, adenine nucleotide content, membrane integrity and oxygen consumption. A series of chlorophenols inhibited sperm motion (motility and velocity) in a concentration-dependent manner. This could be determined quantitatively and reproducibly by means of videomicrography and automatic computer image analysis. The sperm immobilizing potency increased with increasing chlorination and was positively correlated with lipophilicity. Concentrations which reduced the percentage of moving sperm to 50% of controls ranged from 43 microM for pentachlorophenol (PCP) to 1440 microM for 4-monochlorophenol (4-MCP). Determinations of adenine nucleotides and percentages of viable cells revealed qualitative differences between the action of PCP and the lower chlorinated phenols. While the latter decreased the total adenine nucleotide contents and the percentage of unstained cells in parallel to motion inhibition, no such changes occurred after exposure to immobilizing concentrations of PCP. Penta-, tetra- and trichlorinated phenols stimulated cellular respiration, indicating their uncoupling activity, at concentrations lower than those necessary for motion inhibition. The results indicate that bovine spermatozoa may become a useful in vitro model for the toxicological evaluation of chemicals providing quantitative as well as qualitative data.

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Section: Discussionmentioning

confidence: 98%

Bovine spermatozoa as anin vitro model for studies on the cytotoxicity of chemicals: effects of chlorophenols

1989

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“…Consequently, the yield is extremely low. Primarily, this method was used for testing the motility of sperm populations treated with various pharmacological agents, but not as a preparation method for assisted reproduction [91]. …”

Section: Sperm Separation Techniquesmentioning

confidence: 99%

Sperm preparation for ART

Henkel

Schill

2003

Reprod Biol Endocrinol

416

175

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The onset of clinical assisted reproduction, a quarter of a century ago, required the isolation of motile spermatozoa. As the indication of assisted reproduction shifted from mere gynaecological indications to andrological indications during the years, this urged andrological research to understand the physiology of male germ cell better and develop more sophisticated techniques to separate functional spermatozoa from those that are immotile, have poor morphology or are not capable to fertilize oocytes. Initially, starting from simple washing of spermatozoa, separation techniques, based on different principles like migration, filtration or density gradient centrifugation evolved. The most simple and cheapest is the conventional swim-up procedure. A more sophisticated and most gentle migration method is migration-sedimentation. However, its yield is relatively small and the technique is therefore normally only limited to ejaculates with a high number of motile spermatozoa. Recently, however, the method was also successfully used to isolate spermatozoa for intracytoplasmic sperm injection (ICSI). Sperm separation methods that yield a higher number of motile spermatozoa are glass wool filtration or density gradient centrifugation with different media. Since Percoll® as a density medium was removed from the market in 1996 for clinical use in the human because of its risk of contamination with endotoxins, other media like IxaPrep®, Nycodenz, SilSelect®, PureSperm® or Isolate® were developed in order to replace Percoll®. Today, an array of different methods is available and the selection depends on the quality of the ejaculates, which also includes production of reactive oxygen species (ROS) by spermatozoa and leukocytes. Ejaculates with ROS production should not be separated by means of conventional swim-up, as this can severely damage the spermatozoa. In order to protect the male germ cells from the influence of ROS and to stimulate their motility to increase the yield, a number of substances can be added to the ejaculate or the separation medium. Caffeine, pentoxifylline and 2-deoxyadenosine are substances that were used to stimulate motility. Recent approaches to stimulate spermatozoa include bicarbonate, metal chelators or platelet-activating factor (PAF). While the use of PAF already resulted in pregnancies in intrauterine insemination, the suitability of the other substances for the clinical use still needs to be tested. Finally, the isolation of functional spermatozoa from highly viscous ejaculates is a special challenge and can be performed enzymatically to liquefy the ejaculate. The older method, by which the ejaculate is forcefully aspirated through a narrow-gauge needle, should be abandoned as it can severely damage spermatozoa, thus resulting in immotile sperm.

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“…However, this method did not take into consideration the dilution of lithium by buffer that occurred in the lower chamber during the incubation period. A modified TMMR technique (Raoof et al, 1987) provides exposure of the semen to a constant concentration of lithium throughout the 2 h incubation time. This is achieved by placing in the lower chamber a buffer containing the same concentration of the drug as in the upper chamber.…”

Section: Introductionmentioning

confidence: 99%

“…To examine the effect of lithium on sperm motility, the modified trans-membrane migration ratio (TMMR) method (Raoof et al, 1987) was used. In this modification of the technique, lithium in buffer was put in both the upper and the lower chambers.…”

Section: Introductionmentioning

confidence: 99%

Lithium inhibits human sperm motility in vitro.

Raoof

Pearson

Turner

1989

Brit J Clinical Pharma

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Lithium is a widely prescribed drug used for the treatment of bipolar affective illness. Previous reports on its effects on sperm motility and male fertility are conflicting. The effect of lithium on human sperm motility was examined in vitro using the modified transmembrane migration method. This technique takes account of the dilution of lithium that occurs during the incubation. Lithium inhibits human sperm motility in vitro in concentrations comparable with those reported to be achieved in semen after oral administration.

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A modified trans‐membrane migration method for measuring the effect of drugs on sperm motility.

Cited by 8 publications

References 8 publications

Bovine spermatozoa as anin vitro model for studies on the cytotoxicity of chemicals: effects of chlorophenols

Bovine spermatozoa as anin vitro model for studies on the cytotoxicity of chemicals: effects of chlorophenols

Sperm preparation for ART

Lithium inhibits human sperm motility in vitro.

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