Immunoreactivity of Hu proteins facilitates identification of myenteric neurones in guinea‐pig small intestine

The myenteric plexus of the enteric nervous system controls the movement of smooth muscles in the gastrointestinal system. They extend their axons between two peripheral smooth muscle layers to form a tubular meshwork arborizing the gut wall. How a tubular axonal meshwork becomes established without invading centrally toward the gut epithelium has not been addressed. We provide evidence here that sonic hedgehog (Shh) secreted from the gut epithelium prevents central projections of enteric axons, thereby forcing their peripheral tubular distribution. Exclusion of enteric central projections by Shh requires its binding partner growth arrest specific gene 1 (Gas1) and its signaling component smoothened (Smo) in enteric neurons. Using enteric neurons differentiated from neurospheres in vitro, we show that enteric axon growth is not inhibited by Shh. Rather, when Shh is presented as a point source, enteric axons turn away from it in a Gas1-dependent manner. Of the Gαi proteins that can couple with Smo, G protein α Z (Gnaz) is found in enteric axons. Knockdown and dominant negative inhibition of Gnaz dampen the axon-repulsive response to Shh, and Gnaz mutant intestines contain centrally projected enteric axons. Together, our data uncover a previously unsuspected mechanism underlying development of centrifugal tubular organization and identify a previously unidentified effector of Shh in axon guidance.enteric neuron | axon guidance | chemorepellent | Hedgehog | Gas1

show abstract

“…S2C). Similar results were obtained using anti-HuC/D to label enteric neuron cell bodies (24,25) (Fig. S2C).…”

Section: Gas1 and Smo Are Required In Enteric Neurons To Prevent Centralsupporting

confidence: 72%

Gas1 is a receptor for sonic hedgehog to repel enteric axons

Jin

Martinelli

Zheng

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Neuronal protein HuC/HuD (HuC/D) monoclonal (clone 16A11) antibody labeled neuronal cell nuclei and perikarya, recognizing an epitope within the carboxyterminal domain of HuD (Marusich et al, 1994). Staining pattern of cellular morphology and distribution is the same as previously described (Lin et al, 2002;Murphy et al, 2007). The staining pattern of biotin-conjugated HuC/D monoclonal antibody is identical to that of the unconjugated antibody.…”

Section: Antibody Characterizationmentioning

confidence: 79%

Quantitative assessment of glial cells in the human and guinea pig enteric nervous system with an anti‐Sox8/9/10 antibody

Hoff¹,

Zeller²,

Weyhern³

et al. 2008

J of Comparative Neurology

109

View full text Add to dashboard Cite

Quantitative changes of enteric glia (EGC) have been implicated in gastrointestinal disorders. To facilitate future studies of EGC in human pathology, we aimed to characterize thoroughly glial markers in the human enteric nervous system (ENS) and to compare EGC in man and guinea pig. Whole-mount preparations of the enteric nerve plexuses from human and guinea pig ileum and colon were labeled with antibodies against S100b, glial fibrillary acidic protein (GFAP), and p75NGFR and the transcription factors Sox8/9/10 and neuronally counterstained. Abundant immunoreactivity (IR) for S100b, GFAP, p75NGFR, and Sox8/9/10 was detected in EGC of all studied regions. Although the cytoplasmatic staining pattern of most markers did not permit glial quantification, the nuclear localization of Sox8/9/10-IR allowed to identify and count all EGC individually. In both man and guinea pig, myenteric ganglia were larger and contained more EGC and neurons than submucous ganglia. Furthermore, there were more EGC in the human than in the guinea pig myenteric plexus (MP), glial density was consistently higher in the human ENS, and the glia index (glia:neuron ratio) ranged from 1.3 to 1.9 and from 5.9 to 7.0 in the human submucous plexus (SMP) and MP, respectively, whereas, in guinea pig, the glia index was 0.8-1.0 in the SMP and 1.7 in the MP. The glia index was the most robust quantitative descriptor within one species. This is a comprehensive set of quantitative EGC measures in man and guinea pig that provides a basis for pathological assessment of glial proliferation and/or degeneration in the diseased gut.

show abstract

“…2A). The specific neuronal localization was confirmed by double labeling with anti-HuC/HuD neuronal protein mouse monoclonal antibody that has been shown to label all enteric neurons Lin et al, 2002) (Fig. 2C).…”

Section: Identification Of the Submucosal Bmentioning

confidence: 90%

Action of Bradykinin in the Submucosal Plexus of Guinea Pig Small Intestine

Gao²,

Liu³

et al. 2004

J Pharmacol Exp Ther

View full text Add to dashboard Cite

Intracellular recording methods with "sharp" microelectrodes were used to study actions of bradykinin (BK) on electrical behavior of morphologically identified neurons and the identification and localization of BK receptors in the submucosal plexus of guinea pig small intestine. Exposure to BK depolarized the membrane potential and elevated excitability in submucosal neurons with AH-type electrophysiological behavior and Dogiel II multipolar morphology and in neurons with S-type electrophysiological behavior and uniaxonal morphology. BK-evoked depolarizing responses were associated with increased neuronal input resistance in AH-type neurons and decreased input resistance in S-type neurons. -BK mimicked the excitatory action of BK. Western blot analysis and reverse transcription-polymerase chain reaction confirmed the expression of B 2 receptor protein and mRNA. Binding studies with a fluorescently labeled BK 2 antagonist found expression of B 2 receptors on a majority of the ganglion cells. B 2 receptors occupied 82% of the neurons that expressed immunoreactivity for neuropeptide Y, 75% of the neurons that expressed vasoactive intestinal peptide, 84% of the neurons that expressed substance P, 71% of the neurons that expressed choline acetyltransferase, and all neurons that expressed calbindin immunoreactivity. The results suggest that the B 2 receptor mediates the excitatory action of BK on submucosal plexus neurons. Pathophysiological significance of the excitatory actions on secretomotor neurons might be stimulated mucosal secretion and the secretory diarrhea associated with intestinal inflammatory states.Chemical signaling is a mode of physiological communication between the enteric nervous system (ENS) and the enteric immune system (Wood, 2002). Neural networks in the myenteric and submucosal plexuses of the ENS interact to form an independent integrative nervous system that controls intestinal motility, secretion, and blood flow and interact to organize the activity of each of the three effector systems into functional patterns of digestive behavior (Wood, 1994a;Gershon, 1998;Wood et al., 1999). In so doing, the ENS functions in interactive concert with the enteric immune system. Populations of immune/inflammatory cells (lymphocytes, granulocytes, and mast cells) surround the ENS and expand in number during pathological inflammatory states (e.g., enteric infections, inflammatory bowel disease, and radiation-induced enteritis). Members of the immune/inflammatory cell populations communicate with the ENS in paracrine manner through the release of chemical mediators (Frieling et al., 1994;Wood, 2002;Liu et al., 2003a). Exposure to the inflammatory mediators histamine, mast cell proteases, prostaglandins, leukotrienes, and cytokines modifies both neuronal excitability and neurotransmission (Nemeth et al., 1984;Frieling et al., 1994;Dekkers et al., 1997;Xia et al., 1999;Gao et al., 2002; Liu et al., 2003a,b).The present study was focused on bradykinin (BK) as one of the putative mediators formed during i...

show abstract

Immunoreactivity of Hu proteins facilitates identification of myenteric neurones in guinea‐pig small intestine

Cited by 107 publications

References 32 publications

Gas1 is a receptor for sonic hedgehog to repel enteric axons

Gas1 is a receptor for sonic hedgehog to repel enteric axons

Quantitative assessment of glial cells in the human and guinea pig enteric nervous system with an anti‐Sox8/9/10 antibody

Action of Bradykinin in the Submucosal Plexus of Guinea Pig Small Intestine

Contact Info

Product

Resources

About