Local Regulation of Macrophage Subsets in the Adult Rat Testis: MATERIALS AND METHODS(5)

MATERIALS AND METHODS(5)

For counting and subsequent analysis, the macrophages were separated into total monocytes and macrophages (labeled by combined ED1 and ED2 immunohistochemistry) and total resident macrophages (all ED2+ cells). In the previous study, the ED2+ resident macrophages represented 80% of the total testicular mononuclear phagocyte population. Intratesticular ED1+ED2- monocyte/macrophages and putative ‘‘long-term resident’’ macrophages (ED2+ED1- cells) were calculated by subtraction, as previously described. birth control pills

Hormone RIAs

Serum samples were assayed for LH and FSH by specific RIAs, as previously described. Assay sensitivity was 0.09 ng/ml for LH and 1.5 ng/ml for FSH. Testicular IF and homogenates were assayed for testosterone without extraction using a direct 125I-testosterone RIA, or after extraction with chloroform-hexane using a [3H]testosterone RIA. Extraction recoveries were 78-82%.

MIF ELISA

Testicular IF was assayed for immunoreactive MIF by a sandwich ELISA employing a monoclonal anti-MIF capture antibody, a rabbit polyclonal anti-MIF detector antibody, and a purified recombinant mouse MIF standard, as previously described. Note that some samples were not available for MIF assay because of limitations on sample volumes.

This entry was posted in Adult Rat Testis and tagged Adult Rat Testis, Inhibitory Factor, Macrophage, Seminiferous Tubules, Testosterone.