Labelling of living mammalian spermatozoa with the fluorescent thiol alkylating agent, monobromobimane (MB): immobilization upon exposure to ultraviolet light and analysis of acrosomal status

Journal Article · Sat Mar 01 00:00:00 EST 1986 · J. Exp. Zool.; (United States)

DOI:https://doi.org/10.1002/jez.1402370310· OSTI ID:5576453

Cummins, J M; Fleming, A D; Crozet, N; Kuehl, T J; Kosower, N S; Yanagimachi, R

Living spermatozoa of seven mammalian species were treated with the thiol-alkylating fluorescent labelling compound, monobromobimane (MBBR). MB-labelling alone had no effect on sperm motility, nor on the time course or ability of golden hamster spermatozoa to undergo the acrosome reaction when capacitated in vitro. Exposure of MB-labelled spermatozoa to ultraviolet (UV) light and excitation of the MB fluorochrome resulted in virtually immediate immobilization of the spermatozoa without affecting acrosomal status. UV exposure of unlabelled spermatozoa for up to 30 sec had no effect upon motility. Immobilization of MB-labelled spermatozoa depended on the midpiece being irradiated, as irradiation of the head alone, or of the more distal parts of the principal piece, had little or no effect upon motility. Labelling with MB followed by immobilization of individually selected spermatozoa was most useful for detailing the course and site of occurrence of the acrosome reaction during penetration of the cumulus oophorus by golden hamster spermatozoa in vitro. In these often hyperactivated spermatozoa, precise determination of the acrosomal status could not often otherwise be made due to the difficulty in visualizing the acrosomal region of a vigorously thrashing, hyperactivated spermatozoon. This technique should prove valuable in a variety of studies on sperm motility, capacitation and fertilization, and could also be extended to other cell systems.

• Research Organization: Univ. of Queensland, St. Lucia, Australia
• OSTI ID: 5576453
• Journal Information: J. Exp. Zool.; (United States), Vol. 3
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
SPERMATOZOA
BIOLOGICAL RADIATION EFFECTS
LABELLING
ALKYLATING AGENTS
FERTILITY
HAMSTERS
MAMMALS
ULTRAVIOLET RADIATION
VIABILITY
ANIMALS
BIOLOGICAL EFFECTS
ELECTROMAGNETIC RADIATION
GAMETES
GERM CELLS
RADIATION EFFECTS
RADIATIONS
RODENTS
VERTEBRATES
560121* - Radiation Effects on Cells- External Source- (-1987)