Localization of postresection EGF receptor expression using laser capture microdissection

• Published in: Journal of pediatric surgery Vol. 38; no. 3; pp. 440 - 445
• Main Authors: Knott, Andrew W., Erwin, Christopher R., Profitt, Sherri A., Juno, Russell J., Warner, Brad W.
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA Elsevier Inc 01.03.2003; Elsevier
• Subjects: adaptation; Adaptation, Physiological; Animals; Biological and medical sciences; cell isolation; Cell Separation - instrumentation; Cell Separation - methods; Computer Systems; enterocyte; epidermal growth factor; Frozen Sections; Gastroenterology. Liver. Pancreas. Abdomen; Histocytological Preparation Techniques - instrumentation; Ileum - surgery; Ileum - ultrastructure; Intestinal Mucosa - chemistry; Intestinal Mucosa - ultrastructure; intestine; Lasers; Male; Medical sciences; Mice; Mice, Inbred ICR; Microvilli - chemistry; Microvilli - ultrastructure; Muscle, Smooth - chemistry; Muscle, Smooth - ultrastructure; Other diseases. Semiology; Polymerase Chain Reaction; receptor; Receptor, Epidermal Growth Factor - biosynthesis; Receptor, Epidermal Growth Factor - genetics; RNA, Messenger - isolation & purification; Short Bowel Syndrome - metabolism; Short Bowel Syndrome - pathology; Short gut syndrome; Stomach, duodenum, intestine, rectum, anus; Stomach. Duodenum. Small intestine. Colon. Rectum. Anus; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases; Surgery of the digestive system; Short gut syndrome; receptor; intestine; cell isolation; adaptation; epidermal growth factor; enterocyte; Animal model; Microdissection; Epidermal growth factor; Intestinal disease; Diagnosis; Postoperative; Pathophysiology; Rodentia; Gut; Surgical resection; Experimental study; Gene expression; Vertebrata; Mammalia; Microscopy; Mouse; Animal; Digestive diseases; Technique
• ISSN: 0022-3468; 1531-5037
• DOI: 10.1053/jpsu.2003.50076

Background/Purpose: Epidermal growth factor (EGF) and its receptor (EGFR) are key components in the genesis of adaptation after small bowel resection (SBR). Within intestinal homogenates, EGFR expression is increased after SBR; however, the exact cells responsible for altered EGFR expression are unknown. In this study, laser capture microdissection (LCM) microscopy was used to elucidate the specific cellular compartment(s) responsible for postresection changes in EGFR expression. Methods: Male ICR mice underwent a 50% proximal SBR or sham operation. After 3 days, frozen sections were taken from the remnant ileum. Individual cells from villi, crypt, muscularis, and mesenchymal compartments were isolated by LCM. EGFR mRNA expression for each cell compartment was quantified using real-time polymerase chain reaction (PCR). Results: EGFR expression was increased after SBR within the crypt (2-fold) and muscularis compartments (3-fold). There were no changes detected after SBR in the villus tips or mesenchymal compartments. Conclusions: Increased expression of EGFR in crypts directly correlates with the zone of cell proliferation and supports the hypothesis that EGFR signaling is crucial for the mitogenic stimulus for adaptation. The finding of increased EGFR expression in the muscular compartment is novel and may implicate a role for EGFR as a mediator of the muscular hyperplasia seen after massive SBR. J Pediatr Surg 38:440-445. Copyright 2003, Elsevier Science (USA). All rights reserved.