Alterations in apoptotic markers and egg-specific protein gene expression in the chicken oviduct during pause in laying induced by tamoxifen

The aim of this study was to examine the cell apoptosis, gene expression and activity of caspases 2, 3, 8 and 9, and the mRNA expression of selected egg-specific proteins in the chicken oviduct during pause in egg laying induced by tamoxifen (TMX) treatment. The experiment was carried out on Hy-Line Brown laying hens. The control birds were treated subcutaneously with vehicle (ethanol) and the experimental ones with TMX at a dose of 6 mg/kg of body weight. Hens were treated daily until a pause in egg laying occurred and sacrificed on Day 7 of the experiment. Within the oviductal wall, the highest number of apoptotic cells (TUNEL-positive) was found in the luminal epithelium and the lowest in the stroma. The administration of TMX increased the percentage of apoptotic cells in the magnum, isthmus, and shell gland as well as immunoreactivity for caspases 3 and 9. Real-time PCR analysis revealed the segment-dependent mRNA expression of caspases 2, 3, 8 and 9. Treatment of hens with TMX elevated the level of caspase-2 transcript in the infundibulum, caspases 2, 3 and 8 in the isthmus, and caspase-9 in the shell gland (P < 0.05 - P < 0.001). As shown by fluorometric method TMX caused an increase in the activity of caspases 3 and 8 in the magnum, isthmus and shell gland, and the activity of caspases 2 and 9 in the isthmus and shell gland (P < 0.05 - P < 0.01). The expression of ovalbumin, avidin and ovocleidin-116 mRNAs was decreased (P < 0.05 - P < 0.001), ovocalyxin-36 mRNA level tended to increase, and ovocalyxin-32 expression was not affected by TMX. The results obtained indicate that caspases are involved in the chicken oviduct regression during a pause in laying induced by TMX, and estrogen is involved in the regulation of examined caspase expression and activity. The changes in mRNA transcript levels of some examined egg-specific proteins after TMX treatment suggest that there is a relationship between estrogen action and the expression of these genes.