Sperm-macrophage interaction in the mouse: a quantitative assay in vitro using 111indium oxine-labeled sperm

Journal Article · Tue Dec 01 00:00:00 EST 1987 · Biol. Reprod.; (United States)

DOI:https://doi.org/10.1095/biolreprod37.5.1170· OSTI ID:7240940

Olive, D L; Weinberg, J B; Haney, A F

The role of reproductive tract macrophages in contraception and reproductive failure has become widely recognized. However, in vitro analysis of sperm phagocytosis by macrophages has relied upon a semi-quantitative method of sperm counting that is of limited accuracy and reproducibility. We have developed an assay using murine sperm labeled with /sup 111/indium oxine, and results indicate the labeling to be rapid and efficient. Incorporation of /sup 111/indium into sperm increased the dose and sperm concentration and reached 90% maximal uptake after 15 min incubation, with maximal uptake occurring at 30 min. No decrease in sperm motility was noted with levels of oxine in excess of those required for significant labeling. Maximal labeling efficiency occurred in phosphate-buffered saline (PBS), with Dulbecco's modified Eagle's medium (DMEM) + 10% adult bovine serum (ABS) producing significantly less uptake. Label dissociation was detectable in PBS at room temperature, but at 37 degrees C in DMEM + 10% ABS, loss of label occurred at a rate of 23.5%/h. Addition of labeled sperm to murine macrophage monolayers under optimal conditions resulted in uptake of /sup 111/indium by macrophages, while free label was unincorporated. Results indicated assay specificity for macrophage-limited uptake, with insignificant label uptake by nonphagocytic murine fibroblasts and better sensitivity than sperm counting. Macrophages from Bacillus Calmette-Guerin (BCG)-infected mice resulted in a decrease in sperm uptake. Female macrophages showed greater capacity for sperm uptake than those of the male mouse. These initial studies demonstrated the utility of this model system in enhancing the understanding of sperm-macrophage interaction in the female reproductive tract.

• Research Organization: Duke Univ. Medical Center, Durham, NC (USA)
• OSTI ID: 7240940
• Journal Information: Biol. Reprod.; (United States), Vol. 37:5
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
INDIUM 111
UPTAKE
MACROPHAGES
BIOLOGICAL FUNCTIONS
SPERMATOZOA
PHAGOCYTOSIS
MICE
OXINE
RADIOASSAY
REPRODUCTION
TRACER TECHNIQUES
ANIMAL CELLS
ANIMALS
AROMATICS
AZAARENES
AZINES
BETA DECAY RADIOISOTOPES
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
FUNCTIONS
GAMETES
GERM CELLS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
INDIUM ISOTOPES
INTERMEDIATE MASS NUCLEI
ISOMERIC TRANSITION ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
MINUTES LIVING RADIOISOTOPES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHAGOCYTES
PYRIDINES
QUINOLINES
RADIOISOTOPES
RODENTS
SOMATIC CELLS
VERTEBRATES
551001* - Physiological Systems- Tracer Techniques