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Enrichment of lysozyme-containing cells from the rabbit endocervix by unit gravity sedimentation
Abstract:
Lysozyme is a bacteriolytic enzyme component of the secretory granules of endocervical mucous cells. In order to study the subcellular distribution of this enzyme in specific cell populations, endocervical cells from estrous and 5-day pseudopregnant rabbits were separated by unit gravity sedimentation. The application of this technique to pronase-dispersed endocervical cells from estrous rabbits resulted in the isolation and enrichment of two mucous cell types that were distinguished morphologically into type I and type II cell populations. Lysozyme was identified in both cell types, using an unlabeled antibody enzyme method, and the degree of staining paralleled the number of mucous granules. In the absence of estrogen dominance in 5-day pseudopregnant rabbits, there was a 50% reduction in the number of mucous cells with a concomitant reduction in both the number of secretory granules per cell and the intracellular concentration of lysozyme. In the absence of ovarian steroid hormones, i.e., 15-16 weeks after ovariectomy, endocervical cells were devoid of secretory granules and lysozyme staining was negative. Enriched populations of endocervical cells represent a potential experimental model for studying the hormonal role in the regulation of lysozyme synthesis by specific cell populations.