TY  - JOUR
A1  - Hetz, Susan
A1  - Acikgoez, Ali
A1  - Moll, Corinna
A1  - Jahnke, Heinz-Georg
A1  - Robitzki, Andrea A.
A1  - Metzger, Roman
A1  - Metzger, Marco
T1  - Age-related gene expression analysis in enteric ganglia of human colon after laser microdissection
JF  - Frontiers in Aging Neuroscience
N2  - 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.
KW  - enteric nervous system
KW  - myenteric plexus
KW  - aging
KW  - sodium channels
KW  - laser microdissection
Y1  - 2014
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-118308
SN  - 1663-4365
VL  - 6
ER  - 
TY  - JOUR
A1  - Meir, Michael
A1  - Kannapin, Felix
A1  - Diefenbacher, Markus
A1  - Ghoreishi, Yalda
A1  - Kollmann, Catherine
A1  - Flemming, Sven
A1  - Germer, Christoph-Thomas
A1  - Waschke, Jens
A1  - Leven, Patrick
A1  - Schneider, Reiner
A1  - Wehner, Sven
A1  - Burkard, Natalie
A1  - Schlegel, Nicolas
T1  - Intestinal epithelial barrier maturation by enteric glial cells is GDNF-dependent
JF  - International Journal of Molecular Sciences
N2  - Enteric glial cells (EGCs) of the enteric nervous system are critically involved in the maintenance of intestinal epithelial barrier function (IEB). The underlying mechanisms remain undefined. Glial cell line-derived neurotrophic factor (GDNF) contributes to IEB maturation and may therefore be the predominant mediator of this process by EGCs. Using GFAP\(^{cre}\) x Ai14\(^{floxed}\) mice to isolate EGCs by Fluorescence-activated cell sorting (FACS), we confirmed that they synthesize GDNF in vivo as well as in primary cultures demonstrating that EGCs are a rich source of GDNF in vivo and in vitro. Co-culture of EGCs with Caco2 cells resulted in IEB maturation which was abrogated when GDNF was either depleted from EGC supernatants, or knocked down in EGCs or when the GDNF receptor RET was blocked. Further, TNFα-induced loss of IEB function in Caco2 cells and in organoids was attenuated by EGC supernatants or by recombinant GDNF. These barrier-protective effects were blunted when using supernatants from GDNF-deficient EGCs or by RET receptor blockade. Together, our data show that EGCs produce GDNF to maintain IEB function in vitro through the RET receptor.
KW  - enteric glial cells
KW  - neurotrophic factors
KW  - intestinal epithelial barrier
KW  - GDNF5
KW  - RET6
KW  - inflammatory bowel disease
KW  - enteric nervous system
KW  - gut barrier
KW  - intercellular junctions
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-258913
SN  - 1422-0067
VL  - 22
IS  - 4
ER  -