Central and peripheral effects of L-citrulline on thermal physiology and nitric oxide regeneration in broilers

The mechanism that mediates L-citrulline (L-Cit) hypothermia is poorly understood, and the involvement of nitric oxide signaling has not been fully elucidated. Therefore, this study aimed to determine L-Cit's influence on body temperature and to ascertain the central and peripheral mechanisms associated with this response. Chicks responded to intracerebroventricular (ICV) injection of L-Cit with high and low body temperatures (P < 0.05) depending on the dose tested, for both the surface and rectal temperatures. Peripheral (i.p.) L-Cit injection did not affect body temperature responses. Nitric oxide (NO) concentration and NO synthase (NOS) were influenced with varying doses of L-Cit. Hypothalamic NO was increased at 4 µg L-Cit whereas, plasma iNOS was elevated at 2µg L-Cit treatment. However, i.p. L-Cit did not change the NO content, rather it induced higher (P < 0.05) plasma tNOS and iNOS activity, and further upregulated iNOS and nNOS gene expression in the hypothalamus. In addition, ICV L-Cit potentiated a pro- versus anti-inflammatory milieu with the induction of IL-8, IL-10, and TGFβ (P < 0.05), which may be related to the changes in body temperature. Following ICV L-Cit administration, it was observed that L-Cit caused dose variable changes in the ultrastructure of hypothalamic neurons. The lowest dose was associated with a higher number of dead or degenerating neurons, whereas the highest L-Cit dose had fewer neuronal numbers with larger sizes. Therefore, this study shows that central and peripheral L-Cit administration imposes changes in body temperature, nitric oxide production, and inflammatory responses, in a dose-dependent manner.