Effects of protein deprivation and re-feeding on P2X2 receptors in enteric neurons

Table 1 Characteristics of primary and secondary antibodies

Tissue antigen	Host	Dilution	Code and reference
NOS	Sheep	1:2000	H205
Calbindin	Mouse	1:500	Swant 300
Calretinin	Goat	1:100	CG1 Swant
ChAT	Goat	1:50	Chemicon
Donkey anti-rabbit IgG Alexa 488		1:500	Molecular probes
Donkey anti-sheep IgG Alexa 594		1:100	Molecular probes
Donkey anti-mouse IgG Alexa 594		1:200	Molecular probes

NOS: Nitric oxide synthase; ChAT: Choline acetyltransferase.

Table 2 Body weight and small-intestinal area of pre- and postnatal protein-deprived rats at 42 d postnatal

	N	D	DR
Body weight (g)	160 ± 10	40 ± 18a	100 ± 22
Area of small intestine (cm2)	20.41 ± 3.7	13.3 ± 0.6a	16.5 ± 0.4

^aP < 0.05 vs protein deprived, Tukey’s test for multiple comparisons.

Table 3 Results of the profile area (μm²), major and minor axes of nitric oxide synthase-IR, calbindin-IR, calretinin-IR and choline acetyltransferase-IR neurons in the ileal myenteric and submucosal plexuses of the N, D and DR groups

	N	D	DR
Myenteric plexus
NOS
Profile	240.4 ± 30.7	197.1 ± 25.8	225.8 ± 26.7
Major axes	25.2 ± 2.1	21.7 ± 1.6a	25.1 ± 1.9
Minor axes	12.1 ± 0.6	11.3 ± 0.8	11.4 ± 1.1
Calbindin
Profile	227.7 ± 43.1	223.6 ± 26.3	307.5 ± 58c
Major axes	23.5 ± 2.8	24.1 ± 2.9	27.9 ± 4.6
Minor axes	12.2 ± 1.1	11.9 ± 0.3	14.0 ± 0.8c
Calretinin
Profile	397.7 ± 39.6	259.2 ± 48.8a	331.5 ± 24.5
Major axes	29.3 ± 2.1	22.3 ± 3.5a	27.5 ± 2.7
Minor axes	16.7 ± 0.6	14.1 ± 0.5a	15.1 ± 0.5
ChAT
Profile	229.4 ± 39.4	183.6 ± 39.3	198.7 ± 37.5
Major axes	21.8 ± 2.4	19.4 ± 2.4	20.1 ± 2.5
Minor axes	12.8 ± 1.0	11.7 ± 0.7	12.1 ± 0.8
Submucosal plexus
Calbindin
Profile	244.2 ± 47.6	256.7 ± 34.8	310.7 ± 46.2
Major axes	24.8 ± 1.7	27.4 ± 2.2	27.4 ± 1.7
Minor axes	12.4 ± 1.4	12.2 ± 0.8b	14.5 ± 1.2
Calretinin
Profile	233.8 ± 51.5	200 ± 4.6	242.5 ± 41.6
Major axes	24.1 ± 2.1	20.8 ± 0.5b	25.7 ± 3.1
Minor axes	12.4 ± 1.7	12.1 ± 0.3	12.1 ± 1.4
ChAT
Profile	185.5 ± 18.4	154.3 ± 22.4	175.3 ± 20.0
Major axes	20.1 ± 1.7	17.5 ± 0.8b	18.8 ± 0.7
Minor axes	11.4 ± 0.2	11.1 ± 0.9	11.5 ± 1.1

^aP < 0.05,

^bP < 0.001 vs N and DR groups;

^cP < 0.05 vs N and D groups. Tukey’s test for multiple values, mean ± SE, n = 5. NOS: Nitric oxide synthase; ChAT: Choline acetyltransferase.