Postnatal neural progenitor cells of the enteric nervous system are a potential source for future cell replacement therapies of developmental dysplasia like Hirschsprung’s disease. However, little is known about the molecular mechanisms driving the homeostasis and differentiation of this cell pool. In this work, we conducted Affymetrix GeneChip experiments to identify differences in gene regulation between proliferation and early differentiation of enteric neural progenitors from neonatal mice. We detected a total of 1333 regulated genes that were linked to different groups of cellular mechanisms involved in cell cycle, apoptosis, neural proliferation, and differentiation. As expected, we found an augmented inhibition in the gene expression of cell cycle progression as well as an enhanced mRNA expression of neuronal and glial differentiation markers. We further found a marked inactivation of the canonical Wnt pathway after the induction of cellular differentiation. Taken together, these data demonstrate the various molecular mechanisms taking place during the proliferation and early differentiation of enteric neural progenitor cells.
The enteric nervous system (ENS) is a largely autonomous and highly complex neuronal network found in the gastrointestinal tract. Its two major plexuses are integrated into the layered anatomy of the gut wall and, together with central modulating influences, exert control over gastrointestinal motility, secretion, ion-homeostasis, and immunological mechanisms [
HSCR is hallmarked by an aganglionic distal bowel leading to life-threatening disturbances in intestinal motility. Today’s therapeutic gold standard, the surgical resection of the affected gut segments, is nevertheless associated with problematic long-term outcomes with regard to continence [
Genome-wide gene-expression analyses are a useful tool to examine the genetic programs and cellular interactions and have been widely used to identify potential markers or signalling mechanisms especially in CNS neurospheres or cancer tissues. Further, gene-expression assays have also helped to unravel genetic prepositions associated with HSCR [
Here, we used an Affymetrix microarray analysis to evaluate the genetic expression profile of proliferating murine enteric neural stem cells and its changes during the early differentiation
Cell culturing was conducted as described previously [
Neonatal (P0) C57BL/6 mice without regard to sex were decapitated and the whole gut was removed. After removal of adherent mesentery the longitudinal and circular muscle layers containing myenteric plexus could be stripped as a whole from the small intestine. Tissue was chopped and incubated in collagenase type XI (750 U/mL; Sigma-Aldrich, Taufkirchen, Germany) and dispase II (250
Time schedule of enterosphere culture. The timeline illustrates the schedule of
Single free-floating enterospheres (50 enterospheres/dish) were picked again, washed 3 times in Tris buffer, and transferred into new petri dishes containing either proliferation medium or differentiation medium. Differentiation medium consists of DMEM/F12 containing N2 supplement (1 : 100), penicillin (100 U/mL), streptomycin (100
Enterospheres were proliferated or differentiated for 2 more days, thereby forming the two experimental groups “proliferation” and “differentiation.” The difference in expression between those two groups (differentiation versus proliferation) was successively compared by microarray analysis as described below.
Affymetrix microarray analysis was conducted similar to previously published data in three independent experiments, each with cell cultures prepared from 2 pups from the same litter [
Total RNA of enterospheres of both groups was extracted using the RNeasy Micro Kit (Qiagen). RNA quality was evaluated on Agilent 2100 Bioanalyzer with RNA integrity numbers (RIN) of the samples in this study being in the range from 8 to 10. RIN numbers higher than 8 are considered optimal for downstream application [
Double-stranded cDNA was synthesized from 100 ng of total RNA, subsequently linearly amplified, and biotinylated using the GeneChip WT cDNA Synthesis and Amplification Kit (Affymetrix, Santa Clara, CA, USA) according to the manufacturer’s instructions. 15
Normalization of raw data was performed by the Partek Software 6.6, applying an RMA (Robust Multichip Average) algorithm. Significance was calculated using a
In this study, we investigated the changes of the genetic expression profile that occur during the transition from proliferating to differentiating enteric neural progenitor cells
Analysis of mRNA expression was performed on a GeneChip Mouse Gene 1.0 ST array that determines the expression profile of 28.853 genes. Each gene was interrogated by a median of 27 probes that are spread along the full gene.
In total, the gene chip detected 1454 transcripts to be at least 1.5-fold differentially expressed between proliferating and differentiating enterospheres. 1333 of these transcripts code for already identified proteins. 541 genes were found to be upregulated and 792 genes were found to be downregulated in comparison to proliferating enterospheres (see Supplementary Table
We used the ingenuity pathway analysis software (IPA) and data mining with the science literature search engine
Selected genes related to cell cycle.
Gene | Encoded protein | Fold change | Cell cycle |
---|---|---|---|
AURKA | Aurora kinase A | −2.712 |
|
AURKB | Aurora kinase B | −4.146 |
|
CCNA2 | Cyclin A2 | −4.652 |
|
CCNB1 | Cyclin B1 | −5.752 |
|
CCNB2 | Cyclin B2 | −3.392 |
|
CCND1 | Cyclin D1 | −2.476 |
|
CCND3 | Cyclin D3 | −1.539 |
|
CCNE1 | Cyclin E1 | −1.777 |
|
CCNE2 | Cyclin E2 | −2.847 |
|
CCNF | Cyclin F | −3.211 |
|
CDC6 | Cell division cycle 6 | −1.936 |
|
CDC20 | Cell division cycle 20 | −3.113 |
|
CDC25B | Cell division cycle 25B | −1.636 |
|
CDC25C | Cell division cycle 25C | −2.414 |
|
CDC45 | Cell division cycle 45 | −1.769 |
|
CDCA2 | Cell division cycle associated 2 | −3.461 |
|
CDCA3 | Cell division cycle associated 3 | −3.003 |
|
CDCA5 | Cell division cycle associated 5 | −3.053 |
|
CDCA7L | Cell division cycle associated 7-like | −4.123 |
|
CDCA8 | Cell division cycle associated 8 | −3.467 |
|
CDK1 | Cyclin-dependent kinase 1 | −3.227 |
|
CDK15 | Cyclin-dependent kinase 15 | 1.618 |
|
CDK19 | Cyclin-dependent kinase 19 | 1.619 |
|
CDK5R1 | Cyclin-dependent kinase 5, regulatory subunit 1 (p35) | 1.597 | — |
CENPA | Centromere protein A | −1.895 |
|
CENPE | Centromere protein E, 312 kDa | −4.140 |
|
CENPF | Centromere protein F, 350/400 kDa | −3.927 |
|
CENPI | Centromere protein I | −2.899 |
|
CENPK | Centromere protein K | −2.813 |
|
CENPL | Centromere protein L | −1.864 |
|
CENPM | Centromere protein M | −3.407 |
|
CENPN | Centromere protein N | −2.465 |
|
CENPU | Centromere protein U | −1.624 |
|
SKA1 | Spindle and kinetochore associated complex subunit 1 | −1.532 |
|
SKA2 | Spindle and kinetochore associated complex subunit 2 | −1.582 |
|
SKA3 | Spindle and kinetochore associated complex subunit 3 | −3.490 |
|
SKP2 | S-phase kinase-associated protein 2, E3 ubiquitin protein ligase | −1.845 |
|
SPC25 | SPC25, NDC80 kinetochore complex component | −4.148 |
|
Neural differentiation/development.
Gene | Encoded protein | Fold change |
---|---|---|
|
||
ABCG2 | ATP-binding cassette, subfamily G (WHITE), member 2 (junior blood group) | −1.526 |
ASPM | asp (abnormal spindle) homolog, microcephaly associated ( |
−4.911 |
CDT1 | Chromatin licensing and DNA replication factor 1 | −1.528 |
EGFL7 | EGF-like-domain, multiple 7 | 3.132 |
EPHA2 | EPH receptor A2 | −1.529 |
ETV4 | ets variant 4 | −1.934 |
ETV5 | ets variant 5 | −2.844 |
−2.651 | ||
FABP7 | Fatty acid binding protein 7, brain | −2.095 |
|
||
|
||
ATOH8 | Atonal homolog 8 ( |
1.932 |
AXL | AXL receptor tyrosine kinase | 2.015 |
CRIM1 | Cysteine-rich transmembrane BMP regulator 1 (chordin-like) | 1.999 |
CRLF1 | Cytokine receptor-like factor 1 | 2.382 |
DAB1 | Dab, reelin signal transducer, homolog 1 ( |
−2.297 |
ELK3 | ELK3, ETS-domain protein (SRF accessory protein 2) | −1.613 |
ESCO2 | Establishment of sister chromatid cohesion N-acetyltransferase 2 | −4.767 |
GAP43 | Growth associated protein 43 | 1.613 |
GLDN | Gliomedin | 5.809 |
HMOX1 | Heme oxygenase (decycling) 1 | 1.884 |
KLF9 | Kruppel-like factor 9 | 1.592 |
Lmo3 | LIM domain only 3 | 1.542 |
MAP6 | Microtubule-associated protein 6 | 1.874 |
MYRF | Myelin regulatory factor | 2.527 |
NEUROD4 | Neuronal differentiation 4 | 2.036 |
OLIG1 | Oligodendrocyte transcription factor 1 | 2.660 |
Pvr | Poliovirus receptor | 1.768 |
RGS4 | Regulator of G-protein signaling 4 | 1.955 |
S1PR1 | Sphingosine-1-phosphate receptor 1 | 5.073 |
SOCS2 | Suppressor of cytokine signaling 2 | 2.052 |
2.335 | ||
WIPF1 | WAS/WASL interacting protein family, member 1 | 1.587 |
|
||
|
||
CALB2 | Calbindin 2 | 1.616 |
CNP | 2′,3′-Cyclic nucleotide 3′-phosphodiesterase | 1.732 |
GFAP | Glial fibrillary acidic protein | 2.239 |
MBP | Myelin basic protein | 1.768 |
Mturn | Maturin, neural progenitor differentiation regulator homolog ( |
1.853 |
OMG | Oligodendrocyte myelin glycoprotein | −1.822 |
OPALIN | Oligodendrocytic myelin paranodal and inner loop protein | 39.246 |
PLP1 | Proteolipid protein 1 | 1.630 |
S100B | S100 calcium binding protein B | −1.675 |
TUBB2A | Tubulin, beta 2A class IIa | 1.608 |
TUBB2B | Tubulin, beta 2B class IIb | 1.535 |
TUBB3 | Tubulin, beta 3 class III | 1.976 |
|
||
|
||
ABAT | 4-Aminobutyrate aminotransferase | −1.512 |
ADRA1D | Adrenoceptor alpha 1D | 1.803 |
ADRA2A | Adrenoceptor alpha 2A | 2.900 |
ADRA2B | Adrenoceptor alpha 2B | −2.093 |
CHRM2 | Cholinergic receptor, muscarinic 2 | 1.635 |
CHRM3 | Cholinergic receptor, muscarinic 3 | −1.715 |
CHRNA7 | Cholinergic receptor, nicotinic, alpha 7 (neuronal) | 1.772 |
COMT | Catechol-O-methyltransferase | 1.515 |
DDC | DOPA decarboxylase (aromatic L-amino acid decarboxylase) | 1.711 |
DNM3 | Dynamin 3 | 2.643 |
EPHA5 | EPH receptor A5 | 2.076 |
GRIA3 | Glutamate receptor, ionotropic, AMPA 3 | −1.528 |
GRIA4 | Glutamate receptor, ionotropic, AMPA 4 | −1.997 |
GRIK2 | Glutamate receptor, ionotropic, kainate 2 | −1.565 |
GRM5 | Glutamate receptor, metabotropic 5 | −1.600 |
HTR1B | 5-Hydroxytryptamine (serotonin) receptor 1B, G-protein-coupled | −2.377 |
HTR2B | 5-Hydroxytryptamine (serotonin) receptor 2B, G-protein-coupled | 2.205 |
LRRTM2 | Leucine-rich repeat transmembrane neuronal 2 | 3.665 |
LRRTM3 | Leucine-rich repeat transmembrane neuronal 3 | 2.210 |
NTM | Neurotrimin | 1.820 |
PENK | Proenkephalin | 3.478 |
PRR7 | Proline rich 7 (synaptic) | 1.788 |
SLC10A4 | Solute carrier family 10, member 4 | 1.824 |
1.867 | ||
SLITRK2 | SLIT and NTRK-like family, member 2 | −2.414 |
SLITRK6 | SLIT and NTRK-like family, member 6 | 1.672 |
STON2 | Stonin 2 | 4.054 |
STXBP3 | Syntaxin-binding protein 3 | 1.730 |
Stxbp3b | Syntaxin-binding protein 3B | 1.637 |
SV2C | Synaptic vesicle glycoprotein 2C | 1.929 |
SYT6 | Synaptotagmin VI | 2.571 |
|
||
|
||
ATF3 | Activating transcription factor 3 | 2.579 |
DOK4 | Docking protein 4 | 4.937 |
FEZ2 | Fasciculation and elongation protein zeta 2 (zygin II) | 1.547 |
NAV2 | Neuron navigator 2 | 1.647 |
NRCAM | Neuronal cell adhesion molecule | 2.496 |
PLXNB3 | Plexin B3 | 1.739 |
RGMA | Repulsive guidance molecule family member a | 1.552 |
RNF165 | Ring finger protein 165 | −1.548 |
ROBO2 | Roundabout, axon guidance receptor, homolog 2 ( |
−2.211 |
SEMA3B | Sema domain, immunoglobulin domain (Ig), short basic domain, secreted (semaphorin) 3B | 3.692 |
SEMA3E | Sema domain, immunoglobulin domain (Ig), short basic domain, secreted (semaphorin) 3E | 2.877 |
SEMA4F | Sema domain, immunoglobulin domain (Ig), transmembrane domain (TM) and short cytoplasmic domain, (semaphorin) 4F | 4.891 |
SEMA6A | Sema domain, transmembrane domain (TM), and cytoplasmic domain (semaphorin) 6A | −1.707 |
SRGAP1 | SLIT-ROBO Rho GTPase activating protein 1 | 1.524 |
UNC5B | unc-5 homolog B ( |
−1.927 |
|
||
|
||
ARTN | Artemin | 2.423 |
FGF2 | Fibroblast growth factor 2 (basic) | 2.264 |
FGF5 | Fibroblast growth factor 5 | 7.704 |
GDF10 | Growth differentiation factor 10 | −2.361 |
GDF11 | Growth differentiation factor 11 | 1.604 |
GDNF | Glial cell derived neurotrophic factor | 4.325 |
GFRA3 | GDNF family receptor alpha 3 | 1.707 |
MET | MET protooncogene, receptor tyrosine kinase | 6.680 |
NGFR | Nerve growth factor receptor | 1.728 |
NTRK3 | Neurotrophic tyrosine kinase, receptor, type 3 | −1.575 |
SNX16 | Sorting nexin 16 | 1.641 |
SPHK1 | Sphingosine kinase 1 | 1.704 |
SPRY1 | Sprouty homolog 1, antagonist of FGF signaling ( |
−1.647 |
Differentiation of smooth muscle cells/ICCs.
Gene | Encoded protein | Fold change |
---|---|---|
|
||
ACTA2 | Actin, alpha 2, smooth muscle, aorta | 1.693 |
ACTG2 | Actin, gamma 2, smooth muscle, enteric | 2.336 |
ACTN1 | Actinin, alpha 1 | −1.724 |
AEBP1 | AE binding protein 1 | 2.702 |
AFAP1 | Actin filament associated protein 1 | 1.638 |
ARID5B | AT-rich interactive domain 5B (MRF1-like) | 1.521 |
Cald1 | Caldesmon 1 | −1.535 |
CNN1 | Calponin 1, basic, smooth muscle | 1.652 |
ENG | Endoglin | −1.552 |
ENPP1 | Ectonucleotide pyrophosphatase/phosphodiesterase 1 | −1.522 |
ENPP2 | Ectonucleotide pyrophosphatase/phosphodiesterase 2 | 2.959 |
ENTPD1 | Ectonucleoside triphosphate diphosphohydrolase 1 | 1.636 |
FOSL2 | FOS-like antigen 2 | 2.566 |
GAMT | Guanidinoacetate N-methyltransferase | 1.725 |
MYO1E | Myosin IE | 1.569 |
MYO5A | Myosin VA (heavy chain 12, myoxin) | 1.680 |
MYO7B | Myosin VIIB | 1.710 |
MYO18A | Myosin XVIIIA | 1.994 |
MYPN | Myopalladin | 1.570 |
NEB | Nebulin | 1.569 |
Nebl | Nebulette | 2.378 |
NUP210 | Nucleoporin 210 kDa | −1.838 |
RBM24 | RNA binding motif protein 24 | 1.548 |
SMTN | Smoothelin | −1.778 |
SSPN | Sarcospan | 1.603 |
TAGLN | Transgelin | 2.706 |
|
||
|
||
GUCY1A3 | Guanylate cyclase 1, soluble, alpha 3 | −1.876 |
GUCY1B3 | Guanylate cyclase 1, soluble, beta 3 | −2.008 |
KIT | v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog | −1.798 |
KITLG | KIT ligand | −1.541 |
ECM.
Gene | Encoded protein | Fold change |
---|---|---|
CHSY3 | Chondroitin sulfate synthase 3 | −1.645 |
COL6A5 | Collagen, type VI, alpha 5 | 1.527 |
COL12A1 | Collagen, type XII, alpha 1 | −1.973 |
COL14A1 | Collagen, type XIV, alpha 1 | 6.135 |
COL16A1 | Collagen, type XVI, alpha 1 | 1.666 |
COL18A1 | Collagen, type XVIII, alpha 1 | 1.595 |
COL27A1 | Collagen, type XXVII, alpha 1 | 1.522 |
COLGALT2 | Collagen beta(1-O)galactosyltransferase 2 | −1.564 |
CSPG4 | Chondroitin sulfate proteoglycan 4 | −2.952 |
CSPG5 | Chondroitin sulfate proteoglycan 5 (neuroglycan C) | −1.585 |
CYR61 | Cysteine-rich, angiogenic inducer, 61 | 1.748 |
ECM1 | Extracellular matrix protein 1 | 2.580 |
HSPG2 | Heparan sulfate proteoglycan 2 | 1.923 |
ITGA1 | Integrin, alpha 1 | −1.665 |
ITGA4 | Integrin, alpha 4 (antigen CD49D, alpha 4 subunit of VLA-4 receptor) | −2.324 |
ITGA7 | Integrin, alpha 7 | 4.203 |
ITGA8 | Integrin, alpha 8 | −2.262 |
ITGA11 | Integrin, alpha 11 | 1.762 |
ITGB3 | Integrin, beta 3 (platelet glycoprotein IIIa, antigen CD61) | −5.342 |
ITGB4 | Integrin, beta 4 | 1.567 |
KRT80 | Keratin 80 | 2.833 |
LAMA4 | Laminin, alpha 4 | −1.537 |
LAMA5 | Laminin, alpha 5 | 1.684 |
LOX | Lysyl oxidase | 3.250 |
LOXL4 | Lysyl oxidase-like 4 | 2.427 |
2.417 | ||
MATN2 | Matrilin 2 | 2.570 |
MMP2 | Matrix metallopeptidase 2 (gelatinase A, 72 kDa gelatinase, 72 kDa type IV collagenase) | 1.668 |
MMP9 | Matrix metallopeptidase 9 (gelatinase B, 92 kDa gelatinase, 92 kDa type IV collagenase) | −5.557 |
MMP15 | Matrix metallopeptidase 15 (membrane-inserted) | −2.017 |
MMP16 | Matrix metallopeptidase 16 (membrane-inserted) | −1.634 |
MMP17 | Matrix metallopeptidase 17 (membrane-inserted) | 1.612 |
MMP19 | Matrix metallopeptidase 19 | 3.236 |
MMP28 | Matrix metallopeptidase 28 | 1.956 |
NDST3 | N-deacetylase/N-sulfotransferase (heparan glucosaminyl) 3 | −5.557 |
P4HA1 | Prolyl 4-hydroxylase, alpha polypeptide I | −1.958 |
PLOD3 | Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 3 | 2.250 |
UGDH | UDP-glucose 6-dehydrogenase | 1.529 |
Wnt.
Gene | Encoded protein | Fold change |
---|---|---|
|
||
FZD7 | Frizzled class receptor 7 | −2.271 |
LEF1 | Lymphoid enhancer-binding factor 1 | −2.680 |
LRP5 | Low density lipoprotein receptor-related protein 5 | −1.571 |
LRRK2 | Leucine-rich repeat kinase 2 | 1.677 |
TCF19 | Transcription factor 19 | −2.217 |
F7L1 | Transcription factor 7-like 1 (T-cell specific, HMG-box) | −1.762 |
WNT5A | Wingless-type MMTV integration site family, member 5A | −2.325 |
WNT7B | Wingless-type MMTV integration site family, member 7B | 2.942 |
|
||
|
||
ARL4C | ADP-ribosylation factor-like 4C | 2.179 |
AXIN2 | Axin 2 | 1.744 |
CCND1 | Cyclin D1 | −2.476 |
CSRNP1 | Cysteine-serine-rich nuclear protein 1 | 1.822 |
RACGAP1 | Rac GTPase activating protein 1 | −3.201 |
SPRY2 | Sprouty homolog 2 ( |
−1.771 |
SPRY4 | Sprouty homolog 4 ( |
−2.771 |
WISP1 | WNT1 inducible signaling pathway protein 1 | 2.489 |
|
||
|
||
APOE | Apolipoprotein E | 1.704 |
DKK2 | Dickkopf WNT signaling pathway inhibitor 2 | 1.731 |
EDIL3 | EGF-like repeats and discoidin I-like domains 3 | 2.258 |
FRZB | Frizzled-related protein | 1.938 |
HIC1 | Hypermethylated in cancer 1 | −1.731 |
JADE1 | Jade family PHD finger 1 | −1.656 |
LRP4 | Low density lipoprotein receptor-related protein 4 | 1.979 |
NARF | Nuclear prelamin A recognition factor | −1.699 |
NEDD4L | Neural precursor cell expressed, developmentally downregulated 4-like, E3 ubiquitin protein ligase | 1.588 |
NKD1 | Naked cuticle homolog 1 ( |
3.220 |
NOTUM | Notum pectinacetylesterase homolog ( |
2.631 |
NOV | Nephroblastoma overexpressed | 2.050 |
PRICKLE1 | Prickle homolog 1 ( |
1.536 |
TLE3 | Transducin-like enhancer of split 3 | 1.714 |
TRIB2 | Tribbles pseudokinase 2 | −1.637 |
|
||
|
||
DAAM2 | Dishevelled associated activator of morphogenesis 2 | −1.993 |
PSRC1 | Proline/serine-rich coiled-coil 1 | −2.235 |
TNIK | TRAF2 and NCK interacting kinase | 1.677 |
TRAF4 | TNF receptor-associated factor 4 | −1.673 |
Detected regulatory influences on the canonical Wnt pathway. Scheme of the canonical Wnt pathway. Pointy arrowheads indicate an activating, blunt arrowheads, an inhibitory influence. The fold-change in expression of genes is written under the respective gene acronyms and colours indicate a general upregulation (red) or downregulation (green). For detailed explanation of the signalling cascade and regulated genes, see text.
The proliferation and differentiation of enteric neural progenitor cells during embryonic and postnatal development are controlled by a complex interplay of various intrinsic and extrinsic factors. Their exact timing is crucial for proper migration and proliferation of neural crest cells and for their differentiation into the various neural cell types that compose the complex neural structures of the ENS. Although research in recent years extended our understanding of ENS development and its pathologies [
Our experiment aimed to elucidate gene regulations in enterospheres that occur while ENS progenitor cells leave their proliferative state and begin to differentiate into more defined and specific cell types. The results of the Affymetrix gene expression analysis showed the up- and downregulation of overall 1333 known genes that code for already identified proteins. 171 of these genes could be linked to cell proliferation (Table
We also checked the regulated genes for apoptosis markers to see whether the stop in proliferation was related to cell death (Supplementary Table
To further evaluate the proliferative conditions of cell types present in enterospheres, we focused on different cell specific markers of neural progenitors as well as neurons, glial, or smooth muscle cells. We consider this complex cellular composition of the enterospheres an advantage compared to more purified neural crest derived neurospheres as we are able to capture complex interactions and secretion mechanisms between cell types that might also play an important role
Furthermore, the establishment of neuronal cell communication was strongly regulated. Here, we found an increased expression of genes related to synaptogenesis (LRRTM2 and 3 [
Additionally, we found that regulated genes directly involved in the differentiation of muscle cells and/or enteric pacemaker cells called interstitial cells of Cajal (Table
Moreover, the regulation of 43 extracellular matrix proteins like collagens, integrins, proteoglycans, and matrix metallopeptidases points to a reconstruction of extracellular environment that has been discussed to influence neural stem cell behaviour [
Within the dataset, it was of special interest to find particularly many regulated genes related to the canonical Wnt pathway (Table
Taken together, it is conceivable that canonical Wnt signalling plays a role in the maintenance of the enteric progenitor pool during proliferation and is switched off at the beginning of differentiation conditions. Indeed, our previous gene expression analyses [
This study focused on the changes in gene expression of enteric neural progenitor cells occurring within the first two days of transition from a proliferative state to differentiation
The authors declare that there is no conflict of interests regarding the publication of this paper.
Peter Helmut Neckel and Roland Mohr contributed equally to this work.
The project was supported by a grant from the German Federal Ministry for Education and Research (01GN0967). The authors would like to thank Andrea Wizenmann, Andreas Mack, and Sven Poths for their helpful advice.