Specific locations and amounts of denatured collagen and collagen-specific chaperone HSP47 in the uterine cervices of old cows compared with those of heifers

Abstract

Collagen, the most abundant extra-cellular matrix in the reproductive tract, performs a critical role in pregnancy. Although detecting damaged collagen in tissues is challenging, we recently developed a new in situ detection method using a denatured collagen detection reagent in bovine oviducts and uteri. Utilizing this method, we evaluated the hypothesis that the locations and amounts of denatured collagen in the uterine cervices of old cows are different from those in young heifers as a result of repeated pregnancies and deliveries. We compared damaged collagen in the uterine cervix at the mid-luteal phase between post-pubertal growing nulliparous heifers (22.1 ± 1.0 months old; n = 5) and old multiparous cows (143.1 ± 15.6 months old; 9 ± 1 parities; sacrificed at least 3 months after the last parturition by vaginal delivery; n = 5). Picrosirius red staining showed collagen in almost all parts of the cervices. Expectedly, the amount of damaged collagen was increased in the cervices of old cows. Additionally, we combined in situ detection and fluorescence immunohistochemistry of the collagen-specific molecular chaperone, the 47 kDa heat shock protein (HSP47). Increased HSP47 amounts were observed in the cervices from the old cows, but damaged collagen and HSP47 were not located in the same areas. The age differences were confirmed by western blotting using the anti-HSP47 antibody. These findings revealed the specific location and amounts of denatured collagen in the uterine cervices of old cows compared with those of heifers.