Serotonin-immunoreactive neurons and mast cells in the mouse esophagus suggest involvement of serotonin in both motility control and neuroimmune interactions.

BACKGROUND: Serotonin is a major transmitter in the gastrointestinal tract, but little is known about the serotonergic system in the esophagus. METHODS: The aim of this study was to use multilabel immunofluorescence to characterize serotonin-positive nerve cell bodies and fibers and their relationship with other neuronal and non-neuronal elements in the mouse esophagus. Antibodies against serotonin, vesicular acetylcholine transporter (VAChT), choline acetyltransferase (ChAT), protein gene product 9.5 (PGP 9.5), and α-bungarotoxin (α-BT), were used. KEY RESULTS: Serotonin-containing perikarya represented ∼10% of all PGP 9.5-positive myenteric neurons. Serotonin-positive varicose nerve fibers were found in the lamina muscularis mucosae and present on ∼13% of α-BT-labeled motor endplates in addition to VAChT-immunoreactive motor terminals. As ChAT-positive neurons of the compact formation of the nucleus ambiguus were negative for serotonin, serotonin-positive varicosities on motor endplates are presumed to be of enteric origin. On the other hand, cholinergic ambiguus neurons were densely supplied with serotonin-positive varicosities. The tela submucosa and tunica adventitia contained large numbers of serotonin-positive mast cells, a few of which were in close association with serotonin-positive nerve fibers. CONCLUSIONS & INFERENCES: The mouse esophagus is endowed with a rich serotonin-positive intrinsic innervation, including enteric co-innervation of striated muscles. Serotonin may modulate vagal motor innervation of esophageal-striated muscles not only at the central level via projections of the raphe nuclei to the nucleus ambiguus but also at the peripheral level via enteric co-innervation. In addition, mast cells represent a non-neuronal source of serotonin, being involved in neuroimmune processes.

Galanin modulates vagally induced contractions in the mouse oesophagus.

Nitrergic myenteric neurons co-innervating motor endplates were previously shown to inhibit vagally induced contractions of striated muscle in the rodent oesophagus. Immunohistochemical demonstration of putative co-transmitters, e.g. galanin, in enteric neurons prompted us to study a possible role of galanin in modulating vagally mediated contractions in an in vitro vagus nerve-oesophagus preparation of the mouse. Galanin (1-16) (1-100 nmol L(-1)), in the presence of the peptidase inhibitor, phenanthroline monohydrate, inhibited vagally induced contractions in a concentration-dependent manner (control: 100%; galanin 1 nmol L(-1): 95.6 +/- 1.6%; galanin 10 nmol L(-1): 57.3 +/- 6.5%; galanin 100 nmol L(-1): 31.2 +/- 8.1%, n = 5). The non-selective galanin receptor antagonist, galantide (100 nmol L(-1)), blocked the inhibitory effect of galanin (10 nmol L(-1)) while the selective non-galanin receptor 1 and galanin receptor 3 antagonists, M871 (1 micromol L(-1)) and SNAP37889 (100 nmol L(-1)), respectively, and the nitric oxide synthase inhibitor, NG-nitro-l-arginine methyl ester (L-NAME) (200 micromol L(-1)), failed to affect this galanin-induced response. Simultaneous application of galantide (100 nmol L(-1)) and L-NAME (200 micromol L(-1)) significantly reduced the inhibitory effect of capsaicin (30 mumol L(-1)) on vagally induced contractions when compared with its effect in the presence of L-NAME alone or in combination with the selective galanin receptor 2 or 3 antagonists. An inhibitory effect of piperine on vagally induced contractions was reduced neither by galantide nor by L-NAME. Immunohistochemistry revealed galanin immunoreactive myenteric neurons and nerve fibres intermingling with cholinergic vagal terminals at motor endplates. These data suggest that galanin from co-innervating enteric neurons co-operates with nitric oxide in modulating vagally induced contractions in the mouse oesophagus.

Enteric co-innervation of striated muscle fibres in human oesophagus.

Oesophageal striated muscle of several mammalian species receives dual innervation from both vagal motor fibres originating in the brain stem and enteric nerve fibres originating in myenteric ganglia. The aim of this study was to investigate this so-called enteric co-innervation in the human oesophagus. Histochemical and immunohistochemical methods combined with confocal laser scanning microscopy were utilized to study innervation of 14 oesophagi obtained from body donors (age range 47-95 years). In addition, the distribution of striated and smooth muscle in longitudinal and circular layers of the tunica muscularis was studied semiquantitatively. The upper half of the oesophagus was built up of both muscle types with a predominance (>50-60%) of striated muscle, whereas the lower half consisted of smooth muscle only. The majority of motor endplates was compact and ovoid. Enteric nerve fibres on approximately 17% of motor endplates stained for neuronal nitric oxide synthase, vasoactive intestinal polypeptide, galanin and neuropeptide Y and were completely separated from vagal cholinergic nerve terminals. There was remarkable variability of co-innervation rates between striated muscle bundles with some reaching almost 50%. Myenteric neurons representing the putative source of enteric co-innervating nerve fibres, stained for all these markers, which were almost completely colocalized with NADPH-diaphorase. Our study provides evidence for enteric co-innervation of striated muscle in human oesophagus. From these and recent functional results in various rodent species, we suggest that this innervation component represents an integral part of an intramural reflex mechanism for local most likely inhibitory modulation of oesophageal motility.

Tachykinins are involved in local reflex modulation of vagally mediated striated muscle contractions in the rat esophagus via tachykinin NK1 receptors.

The objective of the present study was to investigate the hypothesis of the presence of a local neural reflex modulating the vagally mediated contractions of striated muscle in the rat esophagus and to determine the possible involvement of tachykinins in such a local neural reflex. Electrical stimulation of the vagus nerve evoked twitch contractile responses that were abolished by d-tubocurarine (5 microM). Capsaicin (1-100 microM) inhibited the vagally mediated twitch contractions o f the normal rat esophageal preparations concentration-dependently but not those of the neonatally capsaicin-treated ones. NG-nitro-L-arginine methyl ester (100 microM), a nitric oxide synthase inhibitor, blocked the inhibitory effect of capsaicin and exogenous application of a nitric oxide donor (1 mM) inhibited the vagally mediated twitch contractions. Capsaicin suppressed acetylcholine release from the normal rat esophageal segments evoked by vagus nerve stimulation but not that from the neonatally capsaicin-treated ones. A selective tachykinin NK1 receptor antagonist (0.1 or 1 microM) attenuated the inhibitory effect of capsaicin. However, antagonists of tachykinin NK2, tachykinin NK3 and calcitonin gene-related peptide receptors (1 microM) did not have any effect. A tachykinin NK1 receptor agonist (1 or 5 microM) inhibited the vagally mediated twitch contractions, which was prevented by NG-nitro-L-arginine methyl ester (100 microM). These data suggest that the rat esophagus might have a local neural reflex inhibiting the vagally mediated striated muscle motility, which consists of capsaicin-sensitive sensory neurons and myenteric nitrergic neurons, and that tachykinins might be involved in the neural reflex through tachykinin NK1 receptors.

Vascularization of the area between free grafts and irradiated graft beds in the neck in rats.

Inflammatory lesions of the vascular endothelium after preoperative radiotherapy often cause healing-delayed healing of free flaps in the irradiated graft bed. We investigated changes in neovascularization in the transition area between grafted tissues and irradiated tissues of the graft bed. We irradiated the neck(30 and 50 Gy total dose) in 102 Wistar rats and then grafted a free myocutaneous gracilis flap to the irradiated region of the neck 4 weeks later. We examined histologically the tissues of the graft, the transition area between the graft and the irradiated graft bed, and the graft bed. In contrast to control rats, the tissues in the irradiated animals showed a qualitatively reduced and a more irregular capillary distribution, with substantial fibrosis in the irradiated graft bed. We also found significant differences in vascularization and mean capillary lumen in the transitional zone between graft and graft bed in the irradiated rats compared with controls (P = 0.004 and P < 0.001, respectively). Both number and diameter of capillaries were reduced in the irradiated graft bed tissue. The graft failed to improve vascularization in the transitional zone between graft and irradiated tissue, so we conclude that it is the vascularization status of the bed tissue rather than that of the transplant tissue that is the limiting factor for graft healing.

A strikingly constant ratio exists between Langerhans cells and other epidermal cells in human skin. A stereologic study using the optical disector method and the confocal laser scanning microscope.

Langerhans cells play an important part in the immune surveillance of the human epidermis. Therefore, a certain distribution and numerical relationship to other epidermal cells can be expected. To quantify epidermal Langerhans cells population extensive studies have been performed using two-dimensional quantification methods on vertical sections or epidermal sheet preparations. Whereas methods using vertical sections were complicated considerably by the sampling procedure, the dendritic shape, and the suprabasal, nonrandom distribution of Langerhans cells, epidermal sheet preparations have their limitations regarding the numerical relationship of Langerhans cells to total epidermal cells and the epidermal morphology as such. In order to improve the validity of data the three-dimensional dissector method combined with confocal laser scanning microscopy has been applied to quantify the number of Langerhans cells and other epidermal cell nuclei per volume unit in cryosections of 24 punch biopsies of normal breast skin of eight women. Furthermore, the ratio of Langerhans cells to other epidermal cells, their number per biopsy, and per skin surface area were calculated. To minimize the bias by shrinkage the reference volume was estimated using Cavalieri's principle. A constant ratio of one Langerhans cells to 53 other epidermal cells was identified in breast skin (interindividual correlation coefficient: 0.952, p < 0.0001). Thus, Langerhans cells represent 1.86% of all epidermal cells; however, a wide interindividual range was found for the number of Langerhans cells per mm2 (912-1806; mean +/- SD 1394 +/- 321) and other epidermal cells per mm2 (47,315-104,588; mean +/- SD 73,952 +/- 19,426). This explains the conflicting results achieved by conventional morphometric assessments relating cell numbers to skin surface area, ignoring the varying thickness of the epidermis. The surprisingly constant relationship of Langerhans cells to other epidermal cells stresses the hypothesis of an epidermal Langerhans cells unit where one Langerhans cells seems to be responsible for the immune surveillance of 53 epidermal cells.

Enteric co-innervation of striated muscle fibers in the esophagus: just a "hangover"?

Striated muscle of the esophagus was until recently considered to consist of "classical" skeletal muscle fibers innervated by cholinergic vagal motoneurons. The recently described co-innervation originating from enteric neurons expressing nNOS, VIP, NPY, and galanin added a new dimension of complexity. The aim of this study was to summarize current knowledge about, and to get further hints as to the possible function of enteric co-innervation of striated esophageal muscle fibers. Aldehyde fixed rat esophagi were processed for immunocytochemistry for CGRP or VAChT (to demonstrate vagal motor terminals), nNOS/NADPH-d, VIP, NPY, and galanin (to demonstrate enteric terminals), met-enkephalin, mu opiate receptor, muscarinic receptors m1-3, soluble guanylyl cyclase, and cGMP dependent kinase type I and II. Motor endplates were visualized using fluorochrome tagged alpha-bungarotoxin to label nicotinic receptors, or with AChE histochemistry. Besides light and confocal laser scanning microscopy, immuno electron microscopy was also employed. Up to 80% of motor endplates were co-innervated. In addition to nNOS, VIP, NPY, and galanin, many enteric terminals in esophageal motor endplates expressed met-enkephalin. Some appeared to stain for the muscarinic m(2) receptor. There was prominent immunostaining for the micro opioid receptor in the sarcolemma at both junctional and extrajunctional sites. Immunostaining for soluble guanylyl cyclase was prominent immediately beneath the clusters of nicotinic receptors. Enteric varicosities and vagal terminals intermingled in motor endplates often without intervening teloglial processes. During ontogeny, initially high co-innervation rates were reduced to adult levels in a cranio-caudally progressing manner. We conclude that, in addition to a possible nitrergic, VIP-, NPY-, and galaninergic modulation of neuromuscular transmission by enteric neurons, opioidergic mechanisms could play a role. On the other hand, cholinergic influence on enteric neurons may be exerted also by the nucleus ambiguus via motor endplates, in addition to the input from the dorsal motor nucleus. The observations that enteric nerve fibers contact striated muscle fibers at specialized sites, i.e., motor endplates, and that these contacts appear in an ordered cranio-caudal sequence after cholinergic motor endplates have been established point to a specific function in neuronal control of esophageal muscle rather than to be an unspecific "hangover" from the smooth muscle past of this organ.

Spatial and temporal organization of TrkB expression in the developing musculature of the mouse esophagus.

TrkB expression was investigated immunocytochemically in the developing musculature of mouse esophagus using conventional and confocal laser scanning microscopy. To demonstrate spatial relationships of TrkB immunoreactive cells to striated and smooth muscle fibers we combined TrkB immunocytochemistry with fluorochrome-tagged alpha-bungarotoxin for labeling of nicotinic acetylcholine receptors, and alpha-smooth muscle actin for labeling of smooth muscle cells. At developmental stages E15 to P7, TrkB immunoreactive cells transiently occurred in a transformation zone where striated intermingled with smooth muscle fibers. This transformation zone started in the rostral esophagus at E15, moved caudally, and disappeared between P7 and P10 in the caudal esophagus. The first TrkB-immunoreactive cells appeared in the outer muscle layer at E15. No TrkB-positive cells exhibited acetylcholine receptor clusters or were positive for alpha-smooth muscle actin. A few showed slight alpha-bungarotoxin staining over their entire surface. Taken together, the appearance of TrkB-expressing cells in the transformation zone suggest a role in muscle transdifferentiation. Alternatively, these results, together with recent in vitro data, suggest that TrkB is expressed in a subpopulation of myoblasts in which acetylcholine receptor clustering may be inhibited through a TrkB-mediated pathway.

Histochemical and immunocytochemical study of nitrergic innervation in human nasal mucosa.

Nitric oxide (NO) is a free radical gas that has been found to be produced in neuronal cells by the action of the enzyme brain nitric oxide synthase (bNOS). The aim of this study was to identify NO-containing nerve structures in the human nasal mucosa by localizing bNOS and to find out whether NO production is attached to the parasympathetic system. For this purpose, immunocytochemistry with antibodies to bNOS and neurofilament was performed. Additionally, nicotinamide-adenine dinucleotide phosphate diaphorase (NADPH-d), an enzyme that correlates with the localization of NO synthase, and acetylcholinesterase were visualized in a histochemical double staining technique on frozen sections. The NADPH-d and bNOS reactions were found in axons of nerve bundles and in subepithelial, glandular, and vascular nerve fibers. Arteries showed a distinctly developed nitric innervation, whereas no activity was found in nerve fibers supplying veins. A high coexistence of NADPH-d in parasympathetic nerves could be detected. These findings suggest that NO takes part in the nerve control functions of the human nasal mucosa.

Vagal and spinal afferent innervation of the rat esophagus: a combined retrograde tracing and immunocytochemical study with special emphasis on calcium-binding proteins.

Vagal afferent neurons contain a variety of neurochemical markers and neuroactive substances, most of which are present also in dorsal root ganglion cells. To test for the suitability of the calcium-binding protein calretinin as a specific marker for vagal afferent fibers in the periphery, immunocytochemistry for this protein was combined with retrograde tracing. Nerve fibers in the rat esophagus, as well as vagal and spinal sensory neurons innervating the esophagus, were investigated for co-localization of calretinin with calbindin, calcitonin gene-related peptide, and NADPH diaphorase. The results indicated that calretinin immunocytochemistry demonstrates neuronal structures known as vagal afferent from other studies, in particular intraganglionic laminar endings. A few enteric neurons whose distribution was unrelated to intraganglionic laminar endings also stained for calretinin. Strikingly, calretinin immunoreactivity was absent from spinal afferent neurons innervating the rat esophagus. In intraganglionic laminar endings and nodose ganglion cells calretinin was highly co-localized with calbindin but not with calcitonin gene-related peptide. On the other hand, calbindin was also found in spinal afferents to the esophagus where it was co-localized with calcitonin gene-related peptide. Vagal afferent neurons innervating the esophagus were never positive for NADPH diaphorase. Thus, calretinin appears to be a more specific marker for vagal afferent structures in the esophagus than calbindin, which is expressed by both vagal and spinal sensory neurons. Calretinin immunocytochemistry may be utilized as a valuable tool for investigations of subpopulations of vagal afferents in certain viscera.

Nonvagal origin of galanin-containing nerve terminals innervating striated muscle fibers of the rat esophagus.

We investigated the origin of galanin-positive nerve fibers on motor endplates in rat esophagus using anterograde 1,1'-dioleyl-3,3,3', 3'-tetramethylindocarbocyanine methane sulfonate (DiI) tracing from the nucleus ambiguus combined with galanin immunocytochemistry and calcitonin gene-related peptide immunocytochemistry. To demonstrate spatial relationships of galanin-positive nerve fibers to vagal and enteric nerve fibers on motor endplates, we combined galanin immunocytochemistry with calcitonin gene-related peptide immunostaining for labeling of vagal terminals, and vasoactive intestinal peptide immunoreactivity and NADPH-diaphorase histochemistry for demonstration of enteric nerve fibers. Within fine varicose nerve fibers, galanin was colocalized with vasoactive intestinal peptide and NADPH-diaphorase to a high degree and turned out to be completely separated from calcitonin gene-related peptide-positive or anterogradely DiI-labeled vagal motor terminals. These results indicate that the enteric nervous system is the most important and possibly the only source of galanin-positive nerve terminals on motor endplates in rat esophagus. Galanin may be, in addition to nitric oxide and vasoactive intestinal peptide, a mediator of the enteric coinnervation of striated muscle in this organ.

Spatial relationships of enteric nerve fibers to vagal motor terminals and the sarcolemma in motor endplates of the rat esophagus: a confocal laser scanning and electron-microscopic study.

Enteric co-innervation of motor endplates in the rat esophagus was studied with confocal laser scanning and electron microscopy. Enteric fibers were demonstrated with immunocytochemistry for nitric oxide synthase, vasoactive intestinal peptide or NADPH-diaphorase histochemistry. Vagal motor terminals were identified with calcitonin gene-related peptide (CGRP) immunocytochemistry. Teloglia was stained with immuno- cytochemistry for S100, and TRITC-tagged alpha-bungarotoxin was used to delineate endplate areas in immmunofluorescence preparations. Both confocal imaging and electron microscopy revealed intimate relationships between enteric and vagal terminals on the one hand, and enteric terminals and the sarcolemma on the other. In addition, electron microscopy could point out direct apposition of a significant proportion of enteric varicosities to vagal motor terminals without intervening teloglial processes. These morphological data are compatible with pre- and postsynaptic modulatory effects of enteric neurons on vagal neuromuscular transmission in striated esophageal muscle.

[Endogenously formed nitric oxide in nasal mucosa of the human: detection by nicotinamide-adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry]. / Endogen gebildetes Stickstoffmonoxyd in der Nasenschleimhaut des Menschen: Nachweis mittels Nikotinamid Adenin Dinukleotid Phosphat-Diaphorase (NADPH-d) Histochemie.

BACKGROUND: Nitric oxide (NO) is an intercellular transmitter, both in the central and in the peripheral nervous system. In addition to nerve cells, NO is also produced in epithelial cells of various tissues and in the endothelium. NO is formed by the action of nitric oxide synthase (NOS). There is evidence that NOS can be marked by NADPH-diaphorase (NADPH-d) activity in many cell types. The aim of this study was to identify NOS-positive structures in human nasal mucosa by using NADPH-d-histochemistry. METHODS: Frozen sections from inferior turbinates were fixed with buffered formalin and then treated according to the description of Vincent and Kimura. Additionally, the same sections underwent a double staining procedure for acetylcholinesterase (Karnovski-Roots) to show a correlation with cholinergic nerve structures. RESULTS: Strong reactions were found in the epithelium and in nerve fibres, compared to less NADPH-d activity in seromucous glands and the endothelium of the different vessel types. Singular NADPH-d positive nerves were found within nerve bundles, periarterially, in the subepithelial layer and surrounding glands and their ducts. A frequent localisation of NADPH-d could be detected in parasympathetic nerve fibres. CONCLUSIONS: The occurrence of NADPH-d in epithelial, glandular and nerval structures suggests that NOS takes part in physiological functions and possible pathophysiological processes of the nose. Similar to findings in various other organs this investigation demonstrated that the neurotransmitter NO can also be associated with the parasympathetic nervous system in the human nasal mucosa.

Neuronal and endothelial nitric oxide synthase immunoreactivity and NADPH-diaphorase staining in rat and human pancreas: influence of fixation.

In this study, we wished to clarify the distribution and co-localization of nitric oxide synthase and NA-DPH-diaphorase (NADPH-d) in nerve cells, nerve fibres and parenchymal cells in exocrine and endocrine pancreas, and to assess the influence of fixation on the staining pattern obtained. For this purpose, we applied nitric oxide synthase immunocytochemistry and NADPH-d histochemistry to rat and human pancreas under different fixation conditions. Antibodies to neuronal and endothelial nitric oxide synthase were similarly applied. We found complete co-localization of neuronal nitric oxide synthase and NADPH-d in ganglion cells, and in nerve fibres around acini, excretory ducts, blood vessels and in islets of Langerhans of rat and human pancreas. Immunoreactivity for endothelial nitric oxide synthase was co-localized with NADPH-d in endothelial cells. However, in NADPH-d reactive islet and ductal epithelial cells we could detect neither brain nor endothelial nitric oxide synthase immunoreactivity with any fixation protocol applied. There were marked differences in NADPH-d staining of both neurons and parenchymal cells under different fixation conditions. These results indicate the existence of different types of NADPH-d, which are associated or not associated with nitric oxide synthase(s), and which are differently influenced by various fixation procedures in rat and human pancreas.

Nitrergic innervation of the rat esophagus: focus on motor endplates.

Nitric oxide synthase immunocytochemistry and correlated NADPH-diaphorase histochemistry were utilized to investigate nitrergic innervation of the rat esophagus. Almost all neuronal cell bodies and fibers around blood vessels, and in submucosa and mucosa which were immunoreactive for NOS, also co-stained for NADPH-diaphorase. A combined demonstration of motor endplates with tetramethylrhodamine alpha-bungarotoxin or calcitonin gene-related peptide immunocytochemistry demonstrated a nitrergic co-innervation of striated muscle fibers in all portions of the esophagus. The proportion of endplates co-stained increased from 35% to 78% from the cervical to the abdominal portion of the esophagus. These data indicate a role for NO in neuromuscular transmission in striated muscle of the esophagus.

Nitrergic innervation and nitrergic cells in arteriovenous anastomoses.

Nitrergic innervation and nitrergic epithelioid cells were studied in arteriovenous anastomoses of the tongue, ear, eye, and glomus organ of the finger in different species (rat, rabbit, dog, and man), by means of immunohistochemistry for nitric oxide synthase and enzyme histochemistry utilizing the catalytic activity of this enzyme (the NADPH-diaphorase reaction). Nitrergic perivascular fibers of the tongue were concentrated along the arterial tree and were maximal at the arteriovenous anastomoses in all species. Generally, fewer fibers were located around comparable segments of the episcleral eye vasculature. Only a few nitrergic fibers were found in the canine and rabbit ear, and in the glomus organ of the human finger; however, epithelioid cells in the tunica media of arteriovenous anastomoses of these organs were NADPH-diaphorase-positive and were moderately immunoreactive for nitric oxide synthase. In the epithelioid cells, the reaction product of the NADPH-diaphorase could also be demonstrated by transmission electron microscopy. The epithelioid cells were negative for the panneural and neuroendocrine marker PGP 9.5 confirming the myocytotic nature of these nitrergic cells. Thus, nitric oxide might play a role in mediating the vessel tone of arteriovenous anastomoses via nitrergic nerves or epithelioid cells.

NADPH-diaphorase-positive nerve fibers associated with motor endplates in the rat esophagus: new evidence for co-innervation of striated muscle by enteric neurons.

NADPH-diaphorase histochemistry was combined with demonstration of acetylcholinesterase and immunocytochemistry for calcitonin gene-related peptide to study esophageal innervation in the rat. Most of the myenteric neurons stained positively for NADPH-diaphorase, as did numerous varicose nerve fibers in the myenteric plexus, among striated muscle fibers, around arterial blood vessels, and in the muscularis mucosae. A majority of motor endplates (as demonstrated by acetylcholinesterase histochemistry or calcitonin gene-related peptide immunocytochemistry) were associated with fine varicose NADPH-diaphorase-positive nerve fibers. Analysis of brainstem nuclei, sensory vagal, spinal, and sympathetic ganglia in normal and neonatally capsaicin-treated rats, and comparison with anterogradely labeled vagal branchiomotor, preganglionic and sensory fibers led to the conclusion that NADPH-diaphorase-positive fibers on motor endplates originate in esophageal myenteric neurons. No association of NADPH-diaphorase-positive nerve fibers with motor endplates was found in other organs containing striated muscle. These results suggest extensive, presumably nitrergic, co-innervation of esophageal striated muscle fibers by enteric neurons. Thus, control of peristalsis in the esophagus of the rat may be more complex than hitherto assumed.

Carbonic anhydrase is abundant in fenestrated capillaries of cherry hemangioma.

A strong correlation has been found between carbonic anhydrase (CA) activity and fenestrations in juxtaepithelial capillaries of several tissues, including psoriatic lesions of human skin. In the present study we demonstrate that the majority of the capillaries in cherry hemangiomas are fenestrated and histochemically react CA positively. Obviously the occurrence of CA in these capillaries corresponds to the fenestrations of venous capillaries, which are numerously revealed by electron microscopy. In normal undiseased skin no capillary staining for CA was observed. Therefore in a large proportion of the capillaries of cherry hemangiomas the correlation between fenestrations and CA activity also exists. We suggest that the histochemical demonstration of CA activity might serve as a sensitive and simple marker for fenestrated capillaries in skin tissue.

Carbonic anhydrase--a marker for fenestrated capillaries in psoriasis.

A strong correlation between capillary fenestrations and the demonstration of carbonic anhydrase (CA) has previously been shown. In the present histochemical study we sought to determine whether CA could serve as a marker for fenestrated capillaries in psoriasis. In normal human skin capillary staining for CA was found only in the fenestrated capillaries of the perifollicular and periglandular plexus. In psoriatic skin lesions, however, the intrapapillary capillaries also reacted for CA. From ultrastructural investigations it is known that these capillaries are fenestrated. Our findings have shown that there is a strong correlation between fenestrated and CA-positive capillaries in normal human skin as well as in the lesional skin of psoriatics. Therefore, the demonstration of CA activity may serve as a specific and sensitive marker for fenestrated capillaries in psoriasis using an uncomplicated method, which makes it possible to detect numerous fenestrated capillaries in a single histological section.