Chemical Coding Research Articles

Chemical coding of neurons expressing δ- and κ-opioid receptor and type I vanilloid receptor immunoreactivities in the porcine ileum

Opioid drugs have profound antidiarrheal and constipating actions in the intestinal tract and are effective in mitigating abdominal pain. Mediators of intestinal inflammation and allergy produce increased mucosal secretion, altered bowel motility and pain due to their ability to evoke enteric secretomotor reflexes through primary afferent neurons. In this study, the distribution of delta- and kappa-opioid receptor (DOR and KOR, respectively) immunoreactivities in chemically identified neurons of the porcine ileum was compared with that of the capsaicin-sensitive type 1 vanilloid receptor (VR1). DOR and VR1 immunoreactivities were observed to be highly localized in choline acetyltransferase (ChAT)- and calcitonin gene-related peptide (CGRP)-positive neurons and nerve fibers of the submucosal and myenteric plexuses and both receptors exhibited frequent colocalization. In the inner submucosal plexus, they also were colocalized in substance P (SP)-positive neurons. Neurons in the outer submucosal plexus expressed DOR immunoreactivity alone or in combination with VR1. KOR-immunoreactive neurons were found only in the myenteric plexus; these cells coexpressed immunoreactivity to ChAT, CGRP, vasoactive intestinal peptide (VIP) or nitric oxide synthase (NOS). In addition, some KOR-positive neurons coexpressed immunoreactivities to DOR and VR1. Based on their neurochemical coding, opioid and vanilloid receptor-immunoreactive neurons in the submucosal and myenteric plexuses may include primary afferents and constitute novel therapeutic targets for the palliation of painful intestinal inflammatory, hypersensitivity and dysmotility states.

Distribution and chemical coding of cocaine and amphetamine-regulated transcript (CART) peptide-immunoreactive neurons in the guinea pig bowel and gallbladder

Distribution and chemical coding of cocaine and amphetamine-regulated transcript (CART) peptide-immunoreactive neurons in the guinea pig bowel and gallbladder

Distribution and chemical coding of cocaine and amphetamine-regulated transcript (CART) peptide-immunoreactive neurons in the guinea pig bowel and gallbladder

Distribution and chemical coding of cocaine and amphetamine-regulated transcript (CART) peptide-immunoreactive neurons in the guinea pig bowel and gallbladder

Chemical coding of intrinsic and extrinsic nerves in the guinea pig gallbladder: distributions of PACAP and orphanin FQ.

The complexity of the neural regulation of the gallbladder is reflected by the variety of neuroactive compounds that are found in the intrinsic and extrinsic nerves of the guinea pig gallbladder. The studies reported here used antisera to test for the presence of gallbladder nerves that are immunoreactive for the neuroactive peptides, pituitary adenylyl activating polypeptide (PACAP), and/or orphanin FQ (OFQ, also known as nociceptin). PACAP immunoreactivity was observed in nerve fibers of the paravascular plexus that were also immunoreactive for calcitonin gene-related peptide. These nerve fibers, which are also immunoreactive for substance P, could be followed into the ganglionated plexus. Within the ganglia, a small proportion of neurons was found to be immunoreactive for PACAP; these neurons were also immunoreactive for vasoactive intestinal peptide and nitric oxide synthase. Immunoreactivity for OFQ was observed in the perivascular plexus in nerve fibers that were also immunoreactive for tyrosine hydroxylase. These nerves were previously shown to be immunoreactive for neuropeptide Y. In the ganglionated plexus, immunoreactivity was observed in all gallbladder neurons, as demonstrated by double staining with antiserum directed against the neuron-specific RNA binding protein, Hu. OFQ immunoreactivity was also present in the small catecholaminergic neurons that are observed in a subset of the ganglia. These results further demonstrate the neurotransmitter diversity of the nerves of the gallbladder, and they provide an incentive for studies of the actions of these compounds in the gallbladder wall.

Neurobiologische Grundlagen der Aufmerksamkeit: „Über die Freiheit der Wahl”

Introduction: Attention is viewed as the selective aspect of perception. This selective process is represented as a choice, often but not always constrained by the initial possibilities, motivation and adaptive aims. Fundamental among numerous useful divisions of the concept, is the differentiation of automatic and controlled attention that largely incur serial and parallel processing of information (a classification is proposed in figure 1). Neurophysiology: Selective attention to a stimulus implies that some information is passed on for further processing - this passage depends on the synchronous firing of the relevant neuronal pathways. The EEG sums these and a look at the average after an event shows excitatory and inhibitory components (event-related potentials, ERPs) that relate to the processing stages from registration, inhibiting interference from other processes and stimulus categorisation. Negativities (excitation) can be recorded for deviant detection (automatic, e.g. mismatch negativity, MMN) and attended traces (controlled processing, e.g. negative difference, Nd - see figure 2 for their maturation across adolescence). Gamma, beta and theta frequency bands in the EEG reflect differentially the binding of related features in nearer and further regions of function. Neurotransmitter Roles: The chemical coding of these neural systems with the biogenic amine transmitters allows for a division of labour in the mechanisms necessary for information. These are described in terms of volume control (serotonin), tuning (noradrenaline) and switching (dopamine). The effect of acetylcholine seems to reflect a mechanism by which a stimulus by means of its salience captures automatic processing. (The ascending sensory path, cortical feedback and the mesocorticolimbic source of modulation are schematically illustrated in figure 3) An Anatomy of Attention: Functional magnetic imaging (fMRI) of brain regions activated during attentional processing show that there are 'epicentres' of attentional function superimposed on variable diffusely distributed activity patterns (Mesulam, 1999). Frontal regions exert an executive role, facilitated by cingulate activity in conflict and error-control. Parietal loci on the left and right register temporal and spatial identification, respectively, of events whose relevance can be assessed by comparator mechanisms in the temporal lobe. (Figure 4, compares fMRI and PET activation [frontal, cingulate & intra-parietal sulcus] during an attentional task, Mesulam, 1999.) Choices of what is relevant - selective attention - for the adaptive organisation of response thus reflect a concerted effort within a network of heteromodal cortices based on the basic mechanisms for sorting information arising out of the brainstem.

Distribution and chemical coding of orphanin FQ-immunoreactive neurons in the guinea pig bowel

Distribution and chemical coding of orphanin FQ-immunoreactive neurons in the guinea pig bowel

Distribution, chemical coding and origin of nitric oxide synthase-containing nerve fibres in the guinea pig nasal mucosa

The distribution, chemical coding and origin of nitric oxide synthase (NOS)-containing nerve fibres in the respiratory mucosa of the nasal septum of the guinea pig were examined using nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry and immunohistochemistry. A rich supply of NADPH-d-positive nerve fibres was observed around blood vessels and in nasal glands where nerve fibres frequently penetrated into the epithelia of acini and intralobular ducts. NADPH-d reactivity was also found in the nerve fibres located under or within the respiratory epithelium. Combined immunofluorescence and histochemical staining of the same preparation demonstrated virtually complete overlapping of NOS immunoreactivity and NADPH-d reactivity in nerve fibres, indicating that NADPH-d can be used as a marker for NOS-containing neurons. Double-labelling using antibodies to vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), and calcitonin gene-related peptide (CGRP) revealed that NADPH-d-positive nerve fibres frequently contained VIP or NPY, but not CGRP. Pterygopalatine ganglionectomy significantly reduced the number of NADPH-d-positive nerve fibres innervating the respiratory epithelium as well as blood vessels and nasal glands. Neither superior cervical ganglionectomy nor sensory denervation by capsaicin treatment affected the distribution of NADPH-d-positive fibres. These results indicate that NOS-containing nerve fibres innervating the respiratory epithelium as well as blood vessels and nasal glands in the guinea pig originate mainly from the pterygopalatine ganglion, and suggest that NO may play a significant role as a neurotransmitter and/or neuromodulator in the control of the respiratory epithelium as well as vasculature and nasal glands.

Intrinsic choroidal neurons in the duck eye express galanin

Recently, it has been shown that the choroid of the duck eye harbours approximately 1,000 intrinsic choroidal neurons positive for vasoactive intestinal polypeptide and neuronal nitric oxide synthase. Their connections and functional significance are largely unknown. This study was performed to establish a typical chemical code for these neurons and to define their targets by using immunocytochemistry and confocal laser scanning microscopy. Almost all intrinsic choroidal neurons coexpressed galanin (GAL), vasoactive intestinal polypeptide (VIP), and neuronal nitric oxide synthase (nNOS)/NADPH-diaphorase. A few stained for GAL and/or nNOS only. Among extrinsic ganglia, GAL/VIP/nNOS coexpressing neurons were only found in the pterygopalatine ganglion where they accounted for approximately 30% of the neuronal population. Thus, GAL/VIP/nNOS-positive nerve fibres around branches of the ciliary artery and within the nonvascular smooth muscle stroma of the choroid may originate mainly from intrinsic neurons and to some extent in a subpopulation of pterygopalatine ganglionic neurons exhibiting the same chemical coding. Close contacts of GAL-positive fibres upon intrinsic choroidal neurons may indicate reciprocal connections between them. Thus, intrinsic choroidal neurons may represent peripherally displaced pterygopalatine ganglion neurons forming a local network for regulation of vascular and nonvascular smooth muscle tone in the duck choroid. They may be integrated in the neuronal circuitry controlling intraocular pressure, choroidal thickness, accommodation, and axial bulbus length.

Distribution and chemical coding of orphanin FQ/nociceptin-immunoreactive neurons in the myenteric plexus of guinea pig intestines and sphincter of Oddi.

Longitudinal muscle-myenteric plexus preparations of guinea pig intestines and sphincter of Oddi (SO) were immunostained for orphanin FQ/nociceptin. Orphanin FQ-immunoreactive (OFQ-IR) neurons and nerve fibers were relatively abundant in the SO, duodenum, ileum, cecum, and distal colon, with fewer neurons and nerve fibers observed in the proximal colon. Double staining with antibodies directed against the neuron-specific RNA binding protein Hu revealed that while the numbers of OFQ-IR neurons per ganglion decreased along the gut tube, similar proportions (7-9%) of neurons in these regions were OFQ-IR, whereas <1% of the neurons in the proximal colon were OFQ positive. In the ileum, where 8% of the myenteric neurons were OFQ-IR, all OFQ-IR neurons expressed choline acetyltransferase. In addition, multiple-label immunohistochemistry demonstrated that 58% of the OFQ-IR neurons were calretinin-IR, 52% were substance P-IR, and 28% were enkephalin-IR. Nitric oxide synthase immunoreactivity was observed in about 5% of OFQ-IR neurons, or 0.4% of the total population, and a similar proportion of the OFQ-IR neurons was positive for vasoactive intestinal peptide. No OFQ-IR neurons were immunoreactive for calbindin, somatostatin, or serotonin. These results, combined with previous studies of chemical coding and projection patterns in the guinea pig myenteric plexus, indicate that OFQ-IR is expressed preferentially in excitatory motor neurons projecting to the longitudinal and circular muscle layers, as well as a small subgroup of descending interneurons. Because OFQ is expressed by excitatory motor neurons, and because this peptide inhibits excitatory neurotransmission in the guinea pig ileum, it is likely that OFQ acts through a feedback autoinhibitory mechanism.

Catecholaminergic, cholinergic and peptidergic innervation of gut-associated lymphoid tissue in porcine jejunum and ileum.

With its abundance of neurons and immunocytes, the gut is a potentially important site for the study of the interaction between the nervous and immune systems. Using immunohistochemical techniques, we tested the hypothesis that gut-associated lymphoid tissue in the porcine small intestine might receive catecholaminergic, cholinergic and peptidergic innervation. Antibodies against protein gene product (PGP) 9.5 were employed to detect neuronal membranes; antibodies against tyrosine hydroxylase (TH), type 2 vesicular monoamine transporter (VMAT-2) and choline acetyltransferase (ChAT) were used to detect catecholaminergic and cholinergic neurons; and antibodies to neuromedin U-8 (NMU-8), substance P (SP) and vasoactive intestinal peptide (VIP) were also used. PGP9.5-immunoreactive nerve fibers were observed between jejunal Peyer's patch (PP) follicles and in submucosal ganglia localized at the base of continuous ileal PP. Many ChAT-positive and a few TH-/VMAT-2-immunoreactive neurons or axons adjacent to jejunal and ileal PP were observed. Neurons and fibers from ganglia situated between or at the base of PP follicles manifested robust immunoreactivities to VIP and NMU-8; relatively less SP immunoreactivity was observed at these locations. All neuromedin-U 8-positive neurons observed exhibited immunoreactivity to ChAT as did some VIP-positive neurons. The specific chemical coding of enteric neurons in close apposition to jejunal and ileal PP and the differential localization of neuropeptides within the jejunal and ileal PP are indicative of neuroimmunomodulation at these sites.

Mucosa of the guinea pig gastric corpus is innervated by myenteric neurones with specific neurochemical coding and projection preferences.

The present study identified and characterised myenteric neurones involved in the innervation of the gastric mucosa. We applied retrograde neuronal tracing methods by using the dye DiI (1,1'-didodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorat) in combination with the immunohistochemical demonstration of choline acetyltransferase (ChAT), enkephalin (ENK), neuropeptide Y (NPY), nitric oxide synthase (NOS), substance P (SP), and vasoactive intestinal peptide (VIP). This method showed distinct neurochemical coding of DiI-labelled neurones with projections to the mucosa (mucosa neurones): ChAT/- (indicating the presence of ChAT only, 32%), ChAT/NPY/ +/- VIP (22%), NOS/NPY/ +/- VIP (19%), ChAT/SP/ +/- ENK (12%), NOS/- (indicating the presence of NOS only, 8%), or ChAT/ENK (4.6%). DiI-labelled mucosa neurones did not contain calretinin, serotonin, or somatostatin. All ChAT population had primarily ascending projections, whereas the NOS populations had mainly descending projections. Both were further classified as longitudinally and circumferentially projecting neurones, the latter having projection preferences towards the lesser or greater curvature. All subpopulations exhibited projection preferences. Nitrergic projections primarily arose from cell bodies located at the lesser curvature. ChAT/- projections, which dominated the cholinergic pathway, mainly arose from cell bodies located at the greater curvature. The other major cholinergic pathway with the code ChAT/NPY/ +/- VIP consisted of neurones located mainly at the lesser curvature. The results suggest specific coding of gastric myenteric neurones with projections to the mucosa. Polarised projections consisted of ascending cholinergic and descending nitrergic neurones; the additional presence of NPY/VIP was a prominent feature in both pathways. Chemical coding, polarity, and projection preferences of enteric pathways to the gastric mucosa are remarkably different from those of other regions in the gut.

Multidimensional chemotopic responses to n‐aliphatic acid odorants in the rat olfactory bulb

In an effort to understand the means by which similar chemical odorants are encoded in the mammalian brain, we exposed rats to a homologous series of n-aliphatic acids and mapped the response of the entire olfactory bulb glomerular layer by using a high-resolution [14C]-2-deoxyglucose uptake technique. We found that these similar odorants evoked spatially clustered but distinct responses in the bulb that changed systematically with carbon chain length. In addition to these chemotopic responses, different odorants within the series evoked systematic differences along two other dimensions: amount of deoxyglucose uptake and extent of the glomerular layer showing high activity. Increases along these two dimensions also were correlated with increasing carbon number. The focal glomerular responses were mirrored by responses in deeper bulb layers. Decreasing the odorant concentration decreased the deoxyglucose uptake within focal regions. The focal regions of activity occurred in pairs involving both medial and lateral representations in the bulb, perhaps reflecting the paired medial and lateral projections of olfactory sensory neurons expressing specific types of odorant feature receptor proteins. The observed spatial pattern of response also may explain both the failure of some bulb lesions to interfere with behavioral olfactory responses and the success of other lesions in blocking olfactory responses. These data support a model of parallel, distributed processing of odorants along multiple dimensions. They also support the notion that analyses of the spatial relationships among odorant responses in the olfactory bulb can demonstrate aspects of the mechanism for odor chemical coding. J. Comp. Neurol. 409:529–548, 1999. © 1999 Wiley-Liss, Inc.

Duodenal neurons provide nicotinic fast synaptic input to sphincter of Oddi neurons in guinea pig.

We have investigated the existence of neural connections between the duodenum and the sphincter of Oddi (SO). Stimulation of duodenal myenteric fiber bundles elicited synaptic responses in SO neurons, which included nicotinic fast excitatory postsynaptic potentials (EPSPs), slow EPSPs, and alpha(2)-adrenoreceptor-mediated inhibitory postsynaptic potentials. After 48 h in organ culture, when extrinsic fibers had diminished, only the fast EPSPs persisted. Duodenal mucosal stimulation also elicited nicotinic fast EPSPs in SO neurons. There was no association between the SO neurons that received duodenal input and their chemical coding. A reciprocal projection also exists from the SO to the duodenum. In acute and cultured preparations, duodenal myenteric stimulation caused antidromic responses in 20% of SO neurons. Furthermore, 45.6 +/- 10.5 neurons in SO ganglia were retrogradely labeled from dye application sites in the duodenum. It is proposed that bidirectional neural communication occurs between the duodenum and the SO and that duodenal neurons provide excitatory fast synaptic input to SO neurons through a reflex that can be activated at the duodenal mucosa.

Morphological classifications of enteric neurons -- 100 years after Dogiel

The first differentiation of enteric neurons into three morphological types was done by the russian histologist A. S. Dogiel on the basis of the different shapes and lengths of their dendrites. Although a number of authors considered his results during the following decades, only a division into two types withstood time: type I neurons had one long and several short processes, whereas type II neurons were characterized by several long processes. Some further structural features were discussed but substantial progress was not made until the late 1970s. This stagnation was due to some inaccuracies in Dogiel's descriptions, to the fact that most histologists in this field followed the reticular concept of the nervous system, to the idea that enteric neurons represent no more than a vegetative, postganglionic relay station between the central nervous system and the periphery, and to methodological difficulties. With the application of modern neuroanatomical techniques it was realized that the enteric nervous system contains a considerable number of neuronal subpopulations. The search for morphological correlates of the chemical diversity of enteric neurons was done mainly in the pig and the guinea-pig. In the pig, additional structural features such as axonal projection, distribution of neurons within ganglia, within different plexuses and along the length of the gut, blood supply etc. were included as criteria for further refining neuronal classification. Most of our knowledge about functional features of enteric neurons, e.g. chemical coding, neuronal connectivity, electrophysiological behaviour, was derived from studies in the guinea-pig small intestine. In light of interspecies differences, comparison of findings from different species is mandatory. The search for morphological and functional peculiarities of human enteric neuronal circuitry has to consider all methodological and conceptual advances made within the past 100 years since the pioneering work of Dogiel.

Neurochemical coding of myenteric neurons in the guinea-pig antrum.

Electrophysiological studies of myenteric neurons in the guinea-pig antrum suggest that different neuroactive compounds are involved in synaptic transmission. It is not known what neurotransmitters and neuropeptides are present and to what extent they colocalize. Immunohistochemical stainings were performed on whole-mount preparations of the guinea-pig antrum. Immunoreactivity for neuron-specific enolase was used as a general marker and was set at 100%. There was no overlap between cholinergic and nitrergic neurons, resulting in two separate subpopulations. The presence of choline acetyltransferase immunoreactivity was used to identify the cholinergic subset, which accounted for 56% of the cells. Immunoreactivity for nitric oxide synthase, on the other hand, was displayed in 40.7% of the neurons. Substance-P immunoreactivity was present in 37.4% of the cells and vasoactive intestinal peptide and neuropeptide Y in 21.7% and 28.6%, respectively. Small subsets of neurons had immunoreactivity for serotonin (3.9%), calretinin (6.8%) and calbindin (0.5%). Colocalization studies revealed several subgroups of neurons, containing one or more of the screened markers. Though some similarity is found in the chemical coding of the antrum compared to that of the small intestine and the corpus, remarkable differences can be seen in the occurrence of some subpopulations. Cholinergic neurons are not as predominant as in other parts of the gut, serotonin presence is doubled and some vasointestinal-peptide-positive neurons express substance P. These differences might reflect the highly specialized function of the antrum; however, the exact role of these classes remains to be established.

Multidimensional chemotopic responses to n-aliphatic acid odorants in the rat olfactory bulb

In an effort to understand the means by which similar chemical odorants are encoded in the mammalian brain, we exposed rats to a homologous series of n-aliphatic acids and mapped the response of the entire olfactory bulb glomerular layer by using a high-resolution [14C]-2-deoxyglucose uptake technique. We found that these similar odorants evoked spatially clustered but distinct responses in the bulb that changed systematically with carbon chain length. In addition to these chemotopic responses, different odorants within the series evoked systematic differences along two other dimensions: amount of deoxyglucose uptake and extent of the glomerular layer showing high activity. Increases along these two dimensions also were correlated with increasing carbon number. The focal glomerular responses were mirrored by responses in deeper bulb layers. Decreasing the odorant concentration decreased the deoxyglucose uptake within focal regions. The focal regions of activity occurred in pairs involving both medial and lateral representations in the bulb, perhaps reflecting the paired medial and lateral projections of olfactory sensory neurons expressing specific types of odorant feature receptor proteins. The observed spatial pattern of response also may explain both the failure of some bulb lesions to interfere with behavioral olfactory responses and the success of other lesions in blocking olfactory responses. These data support a model of parallel, distributed processing of odorants along multiple dimensions. They also support the notion that analyses of the spatial relationships among odorant responses in the olfactory bulb can demonstrate aspects of the mechanism for odor chemical coding.

Innervation of the fibro-elastic type of the penis: an immunohistochemical study in the male pig

The occurrence and colocalization of several biologically active neuropeptides, catecholamine-, acetylcholine- or nitric oxide-synthesizing enzymes-tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (D beta H), choline acetyl-transferase (ChAT) and nitric oxide synthase (NOS I), respectively, as well as the vesicular acetylcholine transporter (VAChT) were investigated in the penile glans (GP), corpus and crura (CP), as well as in the retractor penis muscle (RPM) of juvenile and adult boars. Immunohistochemistry revealed that nerves immunoreactive (IR) to TH, D beta H, vasoactive intestinal polypeptide (VIP) and somatostatin (SOM) were the most numerous, followed (in decreasing order of density) by nerves IR to NOS, neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP), galanin (GAL), Leu5-enkephalin (LENK) and ChAT/VAChT. The CP contained the largest number of nerve fibres followed by the RPM, GP and corpus. Enzyme/peptide-containing nerves were associated with both the vascular and non-vascular penile structures. However, differences existed for their density and intrapenile distribution. Nerve terminals IR for different combinations of VIP, GAL or SOM were more frequent than those IR for NOS or CGRP in the non-vascular penile structures while the vasculature and the RPM received a prominent TH/D beta H-, VIP-, SOM- or NOS-IR nerve input. The present data indicate that the porcine penis receives nerve fibres that exhibit diverse chemical codes and that differences in the chemical coding of the nerve fibres may depend on their penile target-structure.

From the cholinergic gene locus to the cholinergic neuron

The cholinergic gene locus (CGL) was first identified in 1994 as the site (human chromosome 10q11.2) at which choline acetyltransferase and a functional vesicular acetylcholine transporter are co-localized. Here, we present recent neuroanatomical, developmental, and evolutionary insights into the chemical coding of cholinergic neurotransmission that have been gleaned from the study of the CGL, and its protein products VAChT and ChAT, which comprise a synthesis-sequestration pathway that functionally defines the cholinergic phenotype.

Identification of cardiovascular pathways in the sympathetic nervous system.

1. Sympathetic autonomic neurons show distinct patterns of expression of a range of neurochemicals that can be detected immunohistochemically. Often, functionally homologous neurons in the autonomic nervous system express identical combinations of substances that serve as a chemical code that allows them to be identified among other autonomic neurons. 2. In the rat stellate ganglion, where many neurons express either immunoreactivity (IR) to neuropeptide Y (NPY) or the calcium-binding protein calbindin, a population of large post-ganglionic neurons found along the medical border of the stellate ganglion, around the origin of the cardiac nerves, expressed intense IR to both substances at all ages examined, from early postnatal to adult. 3. In the heart, in the first few postnatal weeks, many nerve terminals were IR for both NPY and calbindin, but, with increasing age, calbindin-IR was progressively lost from NPY-IR terminals. Nerve terminals IR for both calbindin and NPY were not seen around pulmonary blood vessels or in the trachea or the thymus. 4. Nerve terminals IR for calretinin, another calcium-binding protein, were present in dense pericellular baskets around neurons in the stellate IR for both calbindin and NPY. The terminals also contained nitric oxide synthase (NOS)-IR. 5. It is suggested that the calbindin- and NPY-IR neurons in the stellate ganglion are the post-ganglionic neurons that innervate the heart and that the nerve terminal containing calretinin and NOS-IR that surround them are the cardiac preganglionic terminals. It thus appears possible, in the rat, to identify the sympathetic cardiac pathway arising in the spinal cord and controlling the heart purely on the basis of chemical coding.

The identification and chemical coding of cholinergic neurons in the small and large intestine of the mouse

Background The recent availability of antisera to the vesicular acetylcholine transporter (VAChT) and choline acetyltransferase (ChAT) that demonstrate peripheral cholinergic neurons has made possible the anatomical identification of cholinergic neurons in the enteric nervous system. In this study, we localised cholinergic neurons in the mouse small and large intestine and identified which substances are found colocalised in the cholinergic neurons. Methods Immunohistochemical single and double staining techniques were used on whole mount preparations and frozen sections to examine the localisation and chemical coding of cholinergic neurons in the small and large intestine of the mouse. Cholinergic neurons were identified using antisera to ChAT or VAChT. Results In both the small and large intestine, numerous ChAT-immunoreactive nerve cell bodies were present in the myenteric and submucous ganglia, and ChAT- and VAChT-immunoreactive nerve terminals were abundant in the myenteric and submucous plexuses and the external muscle. Previous studies have identified two major classes of myenteric neurons in the small intestine of the mouse—those containing calretinin plus substance P, and those containing nitric oxide synthase (NOS) plus vasoactive intestinal peptide (VIP). Double-label studies showed that the vast majority of the calretinin/substance P neurons were cholinergic neurons, whereas only a small proportion of the NOS/VIP cells were cholinergic; the noncholinergic NOS/VIP neurons were motor neurons or interneurons, whereas the cholinergic NOS/VIP neurons appeared to be exclusively interneurons. In the small intestine, all of the 5-HT–loaded neurons and a subpopulation of the calbindin neurons were also cholinergic. In the large intestine, there was a pattern of overlaps similar to that found in the small intestine, except that in the large intestine approximately 25% of the calretinin cells were not cholinergic. Only approximately one third of the GABA-loaded neurons in the large intestine were cholinergic. Conclusions Large subpopulations of motor neurons and interneurons in the mouse small intestine are cholinergic neurons. Anat. Rec. 251:185–199, 1998. © 1998 Wiley-Liss, Inc.