Abundance and significance of neuroligin-1 and glutamate in Hirschsprung’s disease

Table 1 Comparison of nutritional status, age and basal metabolic index between Hirschsprung’s disease and non-Hirschsprung’s disease subjects

Item	HSCR(n = 90)	non-HSCR(n = 50)	P value
Age (mo)	7.2 ± 3.15	7.9 ± 2.07	NS
Serum total protein (g/L)	68.8 ± 5.17	70.2 ± 4.29	NS
Serum albumin (g/L)	48.2 ± 7.63	50.5 ± 6.19	NS
Hemoglobin (g/L)	129.1 ± 10.07	131.9 ± 9.89	NS
Blood urea nitrogen (mmol/L)	3.23 ± 1.01	2.91 ± 1.27	NS
Length (cm)	66.2 ± 4.94	67.9 ± 5.18	NS
Weight (kg)	7.8 ± 2.11	8.2 ± 2.75	NS
Basal metabolic index	8.6% ± 0.04%	9.1% ± 0.05%	NS

HSCR: Hirschsprung’s disease; NS: Not significant.

Table 2 Detailed information of antibodies and primers

Antigen	Primary antibody	Dilution	Applications
Neuroligin-1	Goat-anti-human polyclonal	1/100	Detect Nlgn-1 with immunofluorescence on LMMP
Neuroligin-1	Goat-anti-human polyclonal	1/50	Detect Nlgn-1 with Immunohistochemistry on paraffin-embedded sections
Neuroligin-1	Goat-anti-human polyclonal	1/200	Detect Nlgn-1 with Western-blot
Glutamate	Mouse-anti-human monoclonal	1/200	Detect Glu with immunofluorescence on LMMP
Glutamate	Mouse-anti-human monoclonal	1/200	Detect Glu with Immunohistochemistry on LMMP
Glutamate	Mouse-anti-human monoclonal	1/400	Detect Nlgn-1 with Western-blot
β-actin	Rat-anti-human polyclonal	1/2000	Western-blot internal reference
Secondary antibody	Dilution	Applications	Source
Donkey anti-goat Texas Red Secondary	1/500	Label Nlgn-1 with immunofluorescence	ZSGB-BIO China
Goat anti-mouse FITC Secondary	1/200	Label Glu with immunofluorescence	ZSGB-BIO China
Horseradish Peroxidase-conjugated goat-anti-rat IgG	1/500	Detect β-actin with Western-blot	SANTA CRUZ United States
Horseradish Peroxidase-conjugated rabbit-anti-goat IgG	1/1000	Detect Nlgn-1 with Western-blot	SANTA CRUZ United States
Horseradish Peroxidase-conjugated goat-anti-mouse IgG	1/1000	Detect Glu with Western-blot	SANTA CRUZ United States
Primer	Primer sequence (5’→3’)	Annealing temperature (°C)	Product size (bp)
Neuroligin-1	F: GCAAGACCAGAGCAGAGACT	59	314
	R: CACCACCAAAGAATCCAATGTT
β-actin	F: AGCGAGCATCCCCCAAAGTT	60	285
	R: GGGCACGAAGGCTCATCATT

F: Forward primer; R: Reverse primer.