Molecular studies on oestrogen α and progesterone receptors and histomorphometric analysis of canine uteri following aglepristone treatment

Aglepristone, a competitive progesterone antagonist, is successfully used in various progesterone-dependent conditions. This study investigated uterine histomorphometric analysis, and expressions of the oestrogen α receptor (ERα) and progesterone receptor (PR) in uteri of bitches following the single dose of aglepristone treatment. Twelve client-owned healthy diestrous bitches were used in the study. The single dose of aglepristone (Alizine^®, 10 mg/kg) was injected subcutaneously 5 days before ovariohysterectomy in the treatment group (n = 6); bitches without treatment served as a control group (n = 6). Uteri were collected for histomorphometric analysis, ERα and PR gene, and protein expressions studies. The mRNA expressions of ERα and PR were determined by RT-qPCR. Immunohistochemical analysis was used to evaluate the ERα and PR protein expressions using an H-score in five parts of the uterus. The results demonstrated glandular epithelium height significantly decreased (p < .05) and ERα mRNA increased (p < .01) in treated dogs. Of the treated bitches, lower expression levels of ERα were observed in the luminal epithelium, crypt and glandular epithelium, with higher expression in the endometrial stroma and myometrium (p < .05); however, PR expression decreased in the luminal epithelium, crypt and glandular epithelium (p < .01). In conclusion, reduction of the uterine glandular epithelium and ERα mRNA upregulation together with changes in ERα and PR expressions were observed in the treated bitches. However, changes in uterine ERα and PR expressions in the treated bitches depended on tissue layers. The treatment had no effect on serum oestradiol and progesterone levels.     

Differences in the expression of luteinizing hormone and follicle-stimulating hormone receptors in the lower urinary tract between intact and gonadectomised male and female dogs

Receptors for LH (LHR) and FSH (FSHR) are expressed in the canine lower urinary tract (LUT). As gonadectomy results in an increase in plasma LH and FSH, the objective of this study was to determine whether there are any differences in the expression of LHR and FSHR in the LUT between intact and gonadectomised dogs. Four regions of the LUT, i.e. body and neck of the bladder as well as proximal and distal urethra, were collected from 20 healthy dogs (5 intact males, 5 intact anoestrous females, 4 castrated males and 6 spayed females). The mRNA and protein expression of receptors was determined by in situ hybridization and immunohistochemistry, respectively, and assessed semi-quantitatively incorporating both the distribution and the intensity of specific staining. Expression of LHR and FSHR was present in all tissue layers (epithelium, sub-epithelial stroma and muscle) of each region with different levels of the expression. Overall mRNA and protein expression for both LHR and FSHR was significantly (P < 0.001) lower in gonadectomised dogs. Intact dogs had more (P < 0.05) LHR and FSHR mRNA and protein in all tissue layers of the four regions, except for LHR mRNA expression in the sub-epithelial stroma where no differences were observed between the two statuses. Decreases in LHR and FSHR mRNA and protein in gonadectomised dogs appeared to be more consistent in spayed bitches compared to castrated males. Lower expression of LHR and FSHR observed in gonadectomised dogs may adversely affect the normal canine LUT function.