Figure 2

Neuronal and glial cell loss occurs in myenteric plexus ganglia of necrotizing enterocolitis patients. Intestinal specimens were examined from patients with acute necrotizing enterocolitis (NEC; n = 18), from age-matched patients with small bowel atresia (BA; n = 8), from NEC patients undergoing stoma closure (n = 14), and from BA patients undergoing stoma closure (n = 6). (A) Neuronal cell numbers in the myenteric ganglia were counted and neurons per ganglion presented as mean ± standard error of the mean. (B) The number of glial cells per ganglion was extrapolated by subtracting the number of HuC/D-positive neurons from the total numbers of with 4′,6-diamidino-2-phenylindole-labeled nuclei. (C) The ratio of the cytoplasmic area of HuC/D immunoreactive (IR) neurons to myenteric ganglionic area was calculated. (D) Neuronal nitric oxide synthase (nNOS) and HuC/D double-labeled cells in myenteric plexuses were counted and the ratio of nNOS-expressing neurons to total HuC/D stained neurons was calculated. (E) mRNA expression of nNOS as determined by real-time reverse-transcription polymerase chain reaction (RT-PCR). (F) mRNA expression of choline acetyl transferase (chAT) as determined by real-time RT-PCR. *P <0.05; ⁺P <0.01.