title: Age-related gene expression analysis in enteric ganglia of human colon after laser microdissection.
abstract: The enteric nervous system (ENS) poses the intrinsic innervation of the gastrointestinal tract and plays a critical role for all stages of postnatal life. There is increasing scientific and clinical interest in acquired or age-related gastrointestinal dysfunctions that can be manifested in diseases such as gut constipation or fecal incontinence. In this study, we sought to analyze age-dependent changes in the gene expression profile of the human ENS, particularly in the myenteric plexus. Therefore, we used the laser microdissection technique which has been proven as a feasible tool to analyze distinct cell populations within heterogeneously composed tissues. Full biopsy gut samples were prepared from children (4-12 months), middle aged (48-58 years) and aged donors (70-95 years). Cryosections were histologically stained with H&E, the ganglia of the myenteric plexus identified and RNA isolated using laser microdissection technique. Quantitative PCR was performed for selected neural genes, neurotransmitters and receptors. Data were confirmed on protein level using NADPH-diaphorase staining and immunohistochemistry. As result, we demonstrate age-associated alterations in site-specific gene expression pattern of the ENS. Thus, in the adult and aged distal parts of the colon a marked decrease in relative gene expression of neural key genes like NGFR, RET, NOS1 and a concurrent increase of CHAT were observed. Further, we detected notable regional differences of RET, CHAT, TH, and S100B comparing gene expression in aged proximal and distal colon. Interestingly, markers indicating cellular senescence or oxidative stress (SNCA, CASP3, CAT, SOD2, and TERT) were largely unchanged within the ENS. For the first time, our study also describes the age-dependent expression pattern of all major sodium channels within the ENS. Our results are in line with previous studies showing spatio-temporal differences within the mammalian ENS.
authors.affiliation: CELLT Research Group, Translational Centre for Regenerative Medicine, University of Leipzig Leipzig, Germany.
authors.forename: Susan
authors.initials: S
authors.lastname: Hetz
authors.affiliation: Department of General and Visceral Surgery, St. George's Hospital Leipzig, Germany.
authors.forename: Ali
authors.initials: A
authors.lastname: Acikgoez
authors.affiliation: Department of Tissue Engineering and Regenerative Medicine, University Hospital Wuerzburg Wuerzburg, Germany.
authors.forename: Corinna
authors.initials: C
authors.lastname: Moll
authors.affiliation: Division of Molecular biological-biochemical Processing Technology, Center for Biotechnology and Biomedicine (BBZ), University of Leipzig Leipzig, Germany.
authors.forename: Heinz-Georg
authors.initials: HG
authors.lastname: Jahnke
authors.affiliation: Division of Molecular biological-biochemical Processing Technology, Center for Biotechnology and Biomedicine (BBZ), University of Leipzig Leipzig, Germany.
authors.forename: Andrea A
authors.initials: AA
authors.lastname: Robitzki
authors.affiliation: Department of Pediatric Surgery, University of Leipzig Leipzig, Germany.
authors.forename: Roman
authors.initials: R
authors.lastname: Metzger
authors.affiliation: CELLT Research Group, Translational Centre for Regenerative Medicine, University of Leipzig Leipzig, Germany ; Department of Tissue Engineering and Regenerative Medicine, University Hospital Wuerzburg Wuerzburg, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Metzger
keywords: aging; enteric nervous system; laser microdissection; myenteric plexus; sodium channels