title: Abundance and significance of neuroligin-1 and glutamate in Hirschsprung's disease.
abstract: OBJECTIVE
  To investigate the abundance and potential diagnostic significance of neuroligin-1 and glutamate (Glu) in Hirschsprung's disease (HSCR).
  
  
  METHODS
  Ninety children with HSCR and 50 children without HSCR matched for similar nutritional status, age and basal metabolic index were studied. The expression and localization of neuroligin-1 and Glu were assessed using double-labeling immunofluorescence staining of longitudinal muscles with adherent myenteric plexus from the surgically excised colon of children with HSCR. Western blot analysis, quantitative real-time PCR (qRT-PCR) and immunohistochemistry were performed to evaluate the abundance of neuroligin-1 and Glu in different HSCR-affected segments (ganglionic, transitional, and aganglionic segments). Enzyme-linked immunosorbent assay (ELISA) was used to detect and compare serum Glu levels in the long-segment HSCR, short-segment HSCR and non-HSCR samples.
  
  
  RESULTS
  Neuroligin-1 and Glu were co-expressed highest to lowest in the ganglionic, transitional and aganglionic segments based on Western blot (neuroligin-1: 0.177 ± 0.008 vs 0.101 ± 0.006, 0.177 ± 0.008 vs 0.035 ± 0.005, and 0.101 ± 0.006 vs 0.035 ± 0.005, P < 0.005; Glu: 0.198 ± 0.006 vs 0.115 ± 0.008, 0.198 ± 0.006 vs 0.040 ± 0.003, and 0.115 ± 0.008 vs 0.040 ± 0.003, P < 0.005) and qRT-PCR (neuroligin-1: 9.58 × 10(-5) ± 9.94 × 10(-6) vs 2.49 × 10(-5) ± 1.38 × 10(-6), 9.58 × 10(-5) ± 9.94 × 10(-6) vs 7.17 × 10(-6 ±) 1.12 × 10(-6), and 2.49 × 10(-5) ± 1.38 × 10(-6) vs 7.17 × 10(-6) ± 1.12 × 10(-6), P < 0.005). Serum Glu level was the highest to lowest in the non-HSCR, short-type HSCR and long-type HSCR samples based on ELISA (in nmol/μL, 0.93 ± 0.31 vs 0.57 ± 0.25, 0.93 ± 0.31 vs 0.23 ± 0.16, and 0.57 ± 0.25 vs 0.23 ± 0.16, P < 0.005).
  
  
  CONCLUSIONS
  Neuroligin-1 and Glu may represent new markers of ganglion cells, whose expression may correlate with the pathogenesis, diagnosis, differential diagnosis or classification of HSCR.
authors.affiliation: Jian Wang, Wen-Tong Zhang, Hong-Chao Yang, Ai-Wu Li, Department of Pediatric Surgery, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.affiliation: Jian Wang, Wen-Tong Zhang, Hong-Chao Yang, Ai-Wu Li, Department of Pediatric Surgery, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Du
authors.affiliation: Jian Wang, Wen-Tong Zhang, Hong-Chao Yang, Ai-Wu Li, Department of Pediatric Surgery, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China.
authors.forename: Ya-Ru
authors.initials: YR
authors.lastname: Mou
authors.affiliation: Jian Wang, Wen-Tong Zhang, Hong-Chao Yang, Ai-Wu Li, Department of Pediatric Surgery, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China.
authors.forename: Jian-Yi
authors.initials: JY
authors.lastname: Niu
authors.affiliation: Jian Wang, Wen-Tong Zhang, Hong-Chao Yang, Ai-Wu Li, Department of Pediatric Surgery, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China.
authors.forename: Wen-Tong
authors.initials: WT
authors.lastname: Zhang
authors.affiliation: Jian Wang, Wen-Tong Zhang, Hong-Chao Yang, Ai-Wu Li, Department of Pediatric Surgery, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China.
authors.forename: Hong-Chao
authors.initials: HC
authors.lastname: Yang
authors.affiliation: Jian Wang, Wen-Tong Zhang, Hong-Chao Yang, Ai-Wu Li, Department of Pediatric Surgery, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China.
authors.forename: Ai-Wu
authors.initials: AW
authors.lastname: Li
mesh_terms: D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western; D016022:Case-Control Studies; D015816:Cell Adhesion Molecules, Neuronal / Q000032:analysis* / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D003106:Colon / Q000294:innervation*; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay; D005819:Genetic Markers; D018698:Glutamic Acid / Q000032:analysis*; D006627:Hirschsprung Disease / Q000145:classification / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D009197:Myenteric Plexus / Q000737:chemistry*; D011237:Predictive Value of Tests; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D015415:Biomarkers; D015816:Cell Adhesion Molecules, Neuronal; D005819:Genetic Markers; C093438:neuroligin 1; D018698:Glutamic Acid
keywords: Ganglion cells; Glutamate; Hirschsprung’s disease; Neuroligin-1; Pathogenesis