Mechanistic target of rapamycin (mTOR) activation during ruminant mammary development and function : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Animal Science at Massey University, Palmerston North, New Zealand

Abstract

This thesis examines the abundance of total and activated mechanistic target of rapamycin (mTOR) pathway components in the developing and functional ruminant mammary gland. mTOR pathway activation is stimulated by a wide range of intra- and extracellular signals, such as amino acids (AA) and hormones, making the mTOR pathway a potential candidate for the development of intervention strategies designed to increase ruminant lactation potential. Tissues from two trials shown to improve lactation potential; dam-fetal nutrition and exogenous growth hormone (GH) administration during lactation, were used to measure changes in total and activated mTOR pathway protein abundance. Results show mammary glands of d 140 fetal lambs carried by maintenance fed dams and dairy cows administered exogenous GH, had increased abundance of total and activated mTOR and mitogen activated protein kinase (MAPK) pathway proteins. Increased abundance was associated with changes in biochemical indices. In the GH study MAPK pathway activation was stimulated by IGF-1 signaling whilst mTOR pathway activation was proposed to be mediated by AA signalling. Data from the GH study shows, L-arginine a known activator of the mTOR pathway, was the only AA reduced in both plasma and the lactating gland. Upstream factors were not identified for the phenotype observed in the dam-fetal nutrition study, but similar mechanisms were proposed. To elucidate the potential regulation of mTOR pathway activation by L-arginine and examine the effect on milk production, in vitro bovine cell culture models were evaluated. Results show that none of the models evaluated produced a lactating phenotype – a pre-requisite to accurately study the lactating gland in vitro. Finally, this thesis shows L-arginine administration from d 100 to d 140 of pregnancy, in twin bearing ewes had no effect on mTOR protein abundance or activation. However, administration from d 100 to parturition improved maternal gland health. In summary, this thesis associates improved lactation potential with increased total and activated mTOR pathway protein abundance, and the administration of L-arginine during late gestation with improved gland health. These findings provide fundamental knowledge that may lead to the development of novel technologies to increase ruminant gland performance and health