Characterization of the Bovine Mammary Gland Serotonergic System

Abstract

Previous research indicates the presence of a feedback inhibitor of lactation (FIL) in milk (Peaker and Wilde, 1996). Recently, tryptophan hydroxylase I (TPH1), the rate-limiting enzyme in serotonin (5-HT) biosynthesis, was identified in the mouse mammary gland and to be regulated by prolactin (PRL). Furthermore, 5-HT was present in rodent milk and addition of 5-HT to in vitro mammary cultures and in vivo administration of 5-HT to lactating mice reduced milk protein synthesis. Studies were conducted to determine the presence of the enzymatic machinery necessary to produce 5-HT in the bovine mammary gland, the presence of specific 5-HT receptors within the bovine mammary gland, the effects of 5-HT and non-selective and selective receptor antagonists on milk protein gene expression in cultures of primary bovine mammary epithelial cells (BMEC), and to determine the effects of intra-mammary infusions of 5-HT and a non-selective 5-HT receptor antagonist on milk production and composition in lactating dairy cows. Tryptophan hydroxylase I, aromatic amino acid decarboxylase (AADC) and the 5-HT reuptake transporter (SERT) were detected in lactating bovine mammary tissue by qRT-PCR and immunohistochemical staining. Additionally, mRNA for the 5-HT 1B, 2A, 2B, 4 and 7 receptors were detected in bovine mammary tissue as well as BMEC by qRT-PCR and in situ hybridization. Tryptophan hydroxylase I mRNA is in BMEC and up-regulated by PRL. Serotonin down-regulates milk protein gene expression but has not apparent effects on apoptosis and methysergide, ritanserin, SB-224289, and pimozide (receptor antagonists) increased milk protein gene expression in BMEC. Intra-mammary 5-HT infusions decreased overall milk yield in late-lactating dairy cows by 11.1%. Intra-mammary infusions of METH increased overall milk yield by 10.9%. In conclusion, the enzymatic machinery for 5-HT biosynthesis and uptake, as well as receptors involved 5-HT signaling are present in the bovine mammary gland. Furthermore, 5-HT appears to be a FIL in the bovine, but further research regarding its cellular mechanism of action and the location of its receptor populations should be conducted.