Table 1.
Primary and secondary antibodies used in the study
| Antibody | Host | Catalogue number | Dilution | Manufacturer |
|---|---|---|---|---|
| Primary antibody | ||||
| anti-VIP | rabbit | ab22736 | 1:400 | Abcam Limited, Cambridge, UK |
| anti-CART | rabbit | H-003-62 | 1:2,000 | Phoenix Pharmaceuticals, Burlingame, CA, USA |
| anti-nNOS | rabbit | 160870 | 1:200 | Cayman Chemical, Ann Arbor, MI, USA |
| anti-SP | mouse | ab14184 | 1:1,000 | Abcam Limited, Cambridge, UK |
| Secondary antibody | ||||
| anti-mouse/anti-rabbit | goat | DPVB-HRP | RTU | ImmunoLogic, Duiven, the Netherlands |
Fig. 1.
Prenatal effect of β-hydroxy-β-methylbutyrate (HMB) supplementation on the area occupied by cocaine-and-amphetamine-regulated transcript (CART)-immunoreactive (IR), substance P (SP)-IR, vasoactive intestinal peptide (VIP)-IR, and neuronal nitric oxide synthase (nNOS)-IR nerve fibres in the muscular layer of the duodenum, jejunum and ileum (a, b and c), the submucosa of the duodenum, jejunum and ileum (d, e and f), and the mucosa of the duodenum, jejunum and ileum (g, h and i) in newborn piglets prenatally supplemented with HMB and control piglets. * – significant difference between the control and HMB groups at P-value < 0.05; ** – at P-value < 0.01; *** – at P-value < 0.001
Fig. 2.
Representative photomicrographs of the immunohistochemical reactions for cocaine-and-amphetamine-regulated transcript (CART) (A and B), substance P (SP) (C and D), vasoactive intestinal peptide (VIP) (E and F) and neuronal nitric oxide synthase (nNOS) (G and H) in the duodenum (A, B, E and F) and jejunum (C, D, G and H) of control (A, C, E and G) and prenatally HMB-exposed (B, D, F and H) piglets. Arrow – positive reaction. Scale bar = 200 μm