Prostaglandin (PG) E2 plays a crucial role in the endocrine network of canine parturition and we hypothesized that PGE2, 15-hydroxyprostaglandin dehydrogenase (HPGD) and PG-transporter (PGT) might be involved in the development of primary uterine inertia (PUI). We investigated PTGE synthase (PTGES), PTGE receptors 2/4 (PTGER2/4), HPGD and PGT expression on the mRNA- and protein-level in interplacental (IP) and uteroplacental (UP) tissues of bitches presented with dystocia undergoing emergency caesarean section. Groups were formed retrospectively based on strict criteria: PUI (n = 12; small/normal/large litter - PUI-S/N/L: n = 5/4/3), and obstructive dystocia (OD, n = 8). Respective mRNA expressions (ratio) between PUI and OD in IP and UP, between PUI dogs with different litter sizes, between PUI-N and OD in IP, and overall between IP and UP were compared. PTGES, PTGER2, PTGER4, HPGD and PGT mRNA expressions did not differ significantly between PUI and OD in IP or UP. PUI-N PTGES mRNA expression was higher than PUI-S/L (P = 0.0203/P = 0.0186) and OD (P = 0.0314). Higher PTGES (P = 0.0112) and a tendency for higher PTGER2 (P = 0.059) mRNA-expressions were detected in UP versus IP. Other than hypothesized, we did not find a difference in PGE2 production and signaling between PUI and OD, indicating that altered uterine PTGES, PTGER2, PTGER4, HPGD and PGT expression was likely not causative for PUI. However, higher PTGES expression in PUI-N compared to OD might point to a possible role of PGE2 during the course of parturition. Higher PTGES expression in PUI-N compared to PUI-S/L indicates an influence of litter size, the underlying cause and biological relevance of which remain to be clarified.