Collagen and glycosaminoglycan profiles in the canine cervix during different stages of the estrous cycle and in open- and closed-cervix pyometra

Abstract

The extracellular matrix of the cervix that comprises collagen, elastin, proteoglycans and glycosaminoglycans (GAGs) is thought to have an essential role in cervical relaxation. This study investigated the proportion of collagen and smooth muscle as well as the GAGs in cervices obtained from healthy bitches at different stages of the estrous cycle and bitches with open- and closed-cervix pyometra. Cervices were collected after ovariohysterectomy. The proportion of collagen to smooth muscle was determined using Masson's trichrome staining. Alcian blue staining was used to evaluate the relative distribution of cervical GAGs. The proportion of cervical collagen relative to smooth muscle was higher at estrus compared to anestrus (P≤0.05). It was also higher (P≤0.05) in bitches with open- compared to those with closed-cervix pyometra. Overall, hyaluronan (HA) was the predominant GAG in the canine cervix. In the luminal epithelium, the staining intensity for HA was stronger in estrus than in anestrus (P≤0.05), but not in diestrus (P>0.05). On the contrary, the intensity for the combined keratan sulfate (KS) and heparan sulfate (HS) was stronger in anestrus than in estrus and diestrus (P≤0.05). In bitches with pyometra, the staining intensity of the stroma for KS and HS was weaker in open- compared to closed-cervix pyometra (P≤0.05). Collectively, the different profiles of collagen and GAG suggest that the metabolism of both collagen and GAGs in the canine cervix is associated with hormonal statuses during the estrous cycle and cervical patency of bitches with pathological uterine conditions, such as pyometra.

Fig. 1.

Masson’s trichrome staining for blue-stained collagen and red-counterstained smooth muscle in the cervix of healthy bitches at estrus (A), diestrus (B), anestrus (C) and in bitches with open-cervix (D) and closed-cervix pyometra (E). Bar=100 µM.

Fig. 2.

Alcian blue staining in body of the cervical tissue (A–C) of an anestrous bitch. Differences in staining intensity among the 3 varying concentrations of MgCl₂ can be noted; total GAGs at 0.05 MgCl₂ (A), total sulfated GAGs at 0.4 MgCl₂ (B) and highly sulfated GAGs at 0.8 MgCl₂ (C). Cervical cell layers are labeled: luminal epithelium (LE), stroma (S) and smooth muscle (M). Bar=100 µM.