Intermedin and its receptor components in the reproductive systems of the rat and the effect of intermedin on uterine contraction

Abstract

Intermedin (IMD) is a peptide hormone discovered in 2004 belonging to the calcitonin/calcitonin gene-related peptide superfamily. It signals through a Gprotein coupled receptor by the coupling of a calcitonin receptor-like receptor (CRLR) and one of the receptor activity-modifying proteins (RAMPs) 1-3. Due to its similarity to adrenomedullin in structure and functions, IMD is also known as adrenomedullin 2 (ADM2).

Among members of the superfamily, IMD shares the highest degree of homology with ADM, which is a multifunctional vasodilator ubiquitously expressed in various tissues and organs and has been studied by our group for its reproductive functions. It is hypothesized that IMD may be present in the reproductive systems of the rat and exert some effects on reproductive functions.

The objectives of this study were to investigate the expression of IMD and its receptor components in the male and female reproductive systems of the rat, the changes in expression across the oestrous cycle, and its effect on uterine contraction. The gene expression levels of Imd and its receptor components and peptide levels of IMD were measured by RT-PCR and enzyme immunoassay respectively. The effect of IMD on the uterine contraction was studied by the organ bath technique.

Imd mRNA and IMD levels were detected in the testis, epididymis, ventral prostate, coagulating gland, and seminal vesicle of the male rat and the ovary, oviduct, and uterus of the female rat, suggesting possible roles for IMD in both the male and female reproductive systems. In the male, the Imd mRNA levels were the highest in the seminal vesicle but lowest in the testis and the epididymis and IMD peptide levels were the highest in the coagulating gland but lowest in the epididymis. In the female, the Imd mRNA and IMD peptide levels were the highest in the oviduct and the uterus respectively while both the Imd and IMD levels were the lowest in the ovary.

Imd mRNA and IMD levels displayed cyclic changes in various female reproductive tissues across the oestrous cycle. In the ovary, positive immunostaining was detected in the follicles and corpora lutea with more staining in the latter. The Imd mRNA level was significantly higher at prooestrus than dioestrus while the IMD peptide level was significantly higher at metoestrus than dioestrus. In the oviduct, the Imd mRNA level was the lowest at dioestrus but the IMD peptide level was the highest at dioestrus. Positive immunostaining was observed in the ciliated epithelial cells. Uterine Imd mRNA level was the highest at prooestrus while the IMD level was the highest at dioestrus. IMD was found in the luminal and glandular epithelia. IMD significantly reduced the uterine contraction amplitude and frequency but not the basal tone. CGRP receptor antagonist hCGRP8-37 and ADM receptor antagonist hADM22-52 partially abolished the inhibitory effect of IMD on uterine contraction while the IMD specific receptor antagonist hIMD17-47 completely blocked the actions. Enzyme inhibitors of NO (L-NAME) and PI3K (Wortmannin) pathways diminished the IMD-mediated effects on uterine contraction while cAMP/PKA blocker KT5720 had no effect.