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

Cummins J.M., Fleming A.D., Crozet N., Kuehl T.J., Kosower N.S. and Yanagimachi R. (1986) 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 of Experimental Zoology, 237 3: 375-382. doi:10.1002/jez.1402370310


Author Cummins J.M.
Fleming A.D.
Crozet N.
Kuehl T.J.
Kosower N.S.
Yanagimachi R.
Title 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 name Journal of Experimental Zoology   Check publisher's open access policy
ISSN 0022-104X
Publication date 1986-01-01
Sub-type Article (original research)
DOI 10.1002/jez.1402370310
Open Access Status Not yet assessed
Volume 237
Issue 3
Start page 375
End page 382
Total pages 8
Language eng
Subject 1103 Clinical Sciences
Abstract 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, hyper‐activated 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. Copyright
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: Scopus Import - Archived
 
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