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

Abstract

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.

Cadmium, lead, selenium, and zinc in semen of occupationally unexposed men

“…In addition to conventional semen parameters motion activity of the spermatozoa was analysed using computerassisted videomicrography. The equipment employed (Brunner, Munnich, Germany) represented an advanced version of the system described previously (Seibert, 1988;Seibert et al, 1989). The following parameter settings for computer analysis were used in the present study: number of frames analysed: 16; sampling frequency: 25 Hz; minimal number of frames for classification: 9; cell size range: 5-30 pixels; linear velocity threshold: 10 pm s-'.…”

Reprotoxicity of intrauterine insemination and in vitro fertilization-embryo transfer disposables and products: a 4-year survey

“…Table 1 summarizes the range of in vitro and in vivo bioassays currently available for reprotoxicity testing of ART products. These reprotoxicity bioassays test the possible negative influence on the reproductive mechanisms and processes like maturation, motility, and fertilization of mammalian spermatozoa and oocytes of mice, hamsters, rabbits, and bovine (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22). Exposed mouse embryos are being cultured in vitro or transplanted in vivo to test their developmental potential (14,18).…”

Automated analysis of rabbit sperm motility and the effect of chemicals on sperm motion parameters

“…In vitro methods to assess the potential cytotoxic effects of chemicals would be faster and cheaper than toxicology tests utilizing animals and would be in concert with present societal attitude towards animal welfare, since animal use would be greatly reduced. Sperm motility is inhibited or altered by a variety of chemicals, and in vitro chemical cytotoxicity studies with bull spermatozoa have demonstrated a correlation between chemically induced motility inhibition and endpoints of assays that measure cell death (Petzoldt et al, 1985;Seibert et al, 1989;Kemp et al, 1990). These previous studies how-ever, have made no attempts to produce a motion derived toxicological endpoint.…”