Search PubMed⌕ Search

Biomedical subjects

L Thuneberg

Publications and source records attributed to L Thuneberg.

At least 37 records · Page 2Linked to original sources
PubMed1993

Ultrastructure of interstitial cells of Cajal in circular muscle of human small intestine.

BACKGROUND: Interstitial cells of Cajal (ICC) may be important regulatory cells in gut muscle layers. This study examined ICC within the circular muscle of human small intestine. METHODS: Surgically resected, uninvolved intestine was studied by light microscopy and electron microscopy. RESULTS: Muscle lamellae were separated by main septa in continuity with submucosa. Smooth muscle cells ran radially in the septa. Two types of ICC were distinguished. One ICC type had abundant intermediate filaments and smooth cisternae and a discontinuous basal lamina. This ICC type was present in the septa and in the outer third of the circular lamellae. The other ICC type had a complete basal lamina and conspicuous caveolae. This ICC type was observed only in the inner third of the circular lamellae. Both ICC types were close to nerves, but only the latter type formed gap junctions with one another and with muscle cells. Junctions between the two ICC types were not observed. CONCLUSIONS: The arrangement suggests that ICC and radially oriented muscle cells participate in electrical and mechanical coordination of the circular muscle layer of human small intestine.

Adult↗
PubMed1993

Characterization of the outward rectifying potassium channel in a novel mouse intestinal smooth muscle cell preparation.

1. The outward rectifying K+ conductance and underlying single channel behaviour in mouse small intestine (MSI) smooth muscle cells was studied using microelectrode impalement and the patch clamp technique. 2. At 37 degrees C, smooth muscle cells in MSI explants had a resting membrane potential around -65 mV and showed spontaneous electrical and mechanical activity. 3. Under whole-cell voltage clamp, depolarization of smooth muscle cells in the explants evoked a methoxyverapamil (D600)-sensitive, partially inactivating inward current and a non-inactivating outward current. The outward current was also observed in enzymatically dispersed cells from neonatal mouse small intestine. 4. The reversal potential of the outward current as established in tail current experiments was -70.2 mV. Tail currents could be fitted with a single exponential, suggesting the participation of only one population of channels. 5. The outward current was sensitive to 4-aminopyridine (10(-4) M), Ba2+ (1 mM) and to the presence of Cs+ in the pipette, but not to D600 (10(-6) M), or the presence of ATP (1 mM) in the pipette. 6. In the cell-attached patch configuration, a unitary outward current was observed that showed increased activity upon depolarization of the patch. The current-voltage relationship was close to linear with a slope conductance of 186 pS. 7. With normal K+ (6 mM) in the pipette, the extrapolated reversal potential for the unitary current was around -75 mV, while with high K+ (120 mM) the reversal potential was close to 0 mV. 8. Averaging single channel traces recorded under a depolarizing pulse protocol resulted in a trace with similar time characteristics as the outward current observed in the whole-cell configuration. 9. The burst behaviour of the channel was described by a simple model consisting of two closed states, Cf (intraburst closed state) and Cs (interburst closed state) and an open state (O). The rate constants in the model showed differential sensitivity to potential changes, channel blockade by Ba2+ and equimolar K+ conditions. 10. It was concluded that the outward rectifying potassium current in MSI smooth muscle cells is mediated by a 186 pS bursting channel. Voltage dependency and Ba2+ blockade are mainly reflected by changes in the transition rate from the open channel state to the interburst closed state.

4-Aminopyridine↗
PubMed1993

Selective accumulation of methylene blue by interstitial cells of Cajal in canine colon.

The network of interstitial cells of Cajal (ICC) at the submucosal surface of the canine colon was selectively stained by incubation with 15-50 microM methylene blue for 30-45 min. The network was composed of regularly scattered ICC cell bodies interconnected by long processes. Circular muscle cells were unstained. Staining of neurons was limited to one or two axons within bundles. The ICC network had a thickness of a single cell, since no overlapping of ICC cell bodies was observed. The ICC network connected the circular muscle cells at the submucosal surface across the septa which circumferentially divided the circular muscle into lamellae. Methylene blue at 50 microM slightly decreased the resting membrane potential and increased the duration of slow waves, leading to an increase in the force of phasic contractions, with no significant influence on other slow-wave parameters. Methylene blue produced neither electrophysiological nor mechanical effects on circular muscle preparations from which the submuscular ICC network was removed, indicating that the excitatory effects of methylene blue on the full-thickness circular muscle layer were mediated by ICC. In summary, the three-dimensional aspects of the submuscular ICC network can be visualized after selective staining by methylene blue. This staining does not affect physiological characteristics of smooth muscle cells.

Animals↗
PubMed1993

Light- and electron microscopical studies of interstitial cells of Cajal and muscle cells at the submucosal border of human colon.

BACKGROUND: It has been suggested that interstitial cells of Cajal (ICC) at the submucosal border of the colonic circular muscle are pacemaker cells. We studied smooth muscle cells and ICC at the submucosal surface of the circular muscle layer of the normal human colon. EXPERIMENTAL DESIGN: Resected, unaffected specimens from human colon were studied by light microscopy and transmission electron microscopy. RESULTS: Throughout the colon, the inner fourth of each circular-muscle lamella was covered with a layer of 2 to 15 muscle cells (ICMC) with a smaller diameter, more perinuclear organelles, and a greater glycogen content than the outer circular muscle cells. ICMC were interconnected by adherens junctions and close appositions. Small bundles of ICMC were present in the submucosa. ICC were identified in all regions of the colon (ascendens, transversum, and sigmoideum) at the submucosal border, in deeper parts of the submucosa in close contact with smooth muscle bundles as well as in the circular muscle and main septa. ICC had a continuous basal lamina, caveolae, dense bands, thin and intermediate filaments, dense bodies and a well-developed smooth endoplasmic reticulum. Mitochondria and granular endoplasmic reticulum were very abundant. Lipid droplets and glycogen granules were frequent. Thick (myosin) filaments were absent. Close contacts to nerves and gap junctions to other ICC or smooth muscle cells were exceptional. Fibroblast-like cells in the submucosa were rich in granular endoplasmic reticulum and intermediate filaments. They had few dense bands and caveolae. Mitochondria, smooth cisternae and glycogen granules were sparse, cytoplasmic dense bodies and a continuous basal lamina were lacking. Fibroblast-like cells were associated closely with collagen bundles and they had no close contacts with nerves, ICC or muscle cells. CONCLUSIONS: Throughout the normal human colon, submucosal ICC and ICMC are identified and distinguished from other cell types present. Their organization and cytology differ from that of other animal species. The ultrastructure of ICC and ICMC is compatible with important regulatory functions on the circular muscle in the entire human colon.

Aged↗
PubMed1992

Intercellular communication in smooth muscle.

The functioning of a group of cells as a tissue depends on intercellular communication; an example is the spread of action potentials through intestinal tissue resulting in synchronized contraction. Recent evidence for cell heterogeneity within smooth muscle tissues has renewed research into cell coupling. Electrical coupling is essential for propagation of action potentials in gastrointestinal smooth muscle. Metabolic coupling may be involved in generation of pacemaker activity. This review deals with the role of cell coupling in tissue function and some of the issues discussed are the relationship between electrical synchronization and gap junctions, metabolic coupling, and the role of interstitial cells of Cajal in coupling.

Animals↗
PubMed1992

Ultrastructure of interstitial cells of Cajal associated with deep muscular plexus of human small intestine.

Evidence showing that interstitial cells of Cajal have important regulatory functions in the gut musculature is accumulating. In the current study, the ultrastructure of the deep muscular plexus and associated interstial cells of Cajal in human small intestine were studied to provide a reference for identification and further physiological or pathological studies. The deep muscular plexus was sandwiched between a thin inner layer of smooth muscle (one to five cells thick) and the bulk of the circular muscle. Interstitial cells of Cajal in this region very much resembled smooth muscle cells (with a continuous basal lamina, caveolae, intermediate filaments, dense bodies, dense bands, and a well-developed subsurface smooth endoplasmic reticulum), but the arrangement of organelles was clearly different, and cisternae of granular endoplasmic reticulum were abundant. Interstitial cells of Cajal were distinguished from fibroblasts or macrophages in the region. They ramified in the inner zone of the outer division of circular muscle, penetrated the inner-most circular layer, and were also found at the submucosal border. They were in close, synapselike contact with nerve terminals of the deep muscular plexus, and only few gap junctions with other interstitial cells of Cajal or with the musculature were observed. Compared with interstitial cells of Cajal from other mammals, those associated with the deep muscular plexus in the human small intestine more closely resemble smooth muscle cells, and their organization appears more diffuse; however, the ultrastructure and organization of interstitial cells of Cajal is compatible with modulatory actions on the circular muscle also in humans.

Adult↗
PubMed1991

Interstitial cells of Cajal and Auerbach's plexus. A scanning electron microscopical study of guinea-pig small intestine.

Interstitial cells of Cajal (ICC) appear to be involved in the regulation of intestinal motility, probably as pacemaker cells. We investigated the complex organization of ICC associated with Auerbach's plexus of guinea-pig small intestine in the scanning electron microscope. The plexus was exposed by microdissection of zinc iodide/osmic acid stained tissue. After separation of the muscle layers by microdissection alone, the exposed Auerbach's plexus was seen to be covered by a smooth mat of reticular fibrils, thin enough to allow the detailed examination of the intact nerve plexus and interstitial tissue. ICC were distinguished as small, ovoid cell bodies from which 2-5 long, branching, roughly cylindrical processes emerge, associating to form a complex network. Characteristically, ICC processes participated in the formation of small bundles along their course, individual processes passing from one bundle to another. Cell bodies and processes of ICC were intimately associated with tertiary nerves of Auerbach's plexus. Axons were identified and distinguished these from ICC processes by their varicose structure and by th smaller diameters as compared with ICC processes. We found no single axons in our material. The characteristic morphology of ICC clearly distinguished these as a separate cell population different from neurons, glial cells, fibroblasts, and smooth muscle cells. After removal of the mat of reticular fibrils by chemical digestion the detailed organization of the interstitial tissue was preserved. Macrophage-like cells were previously demonstrated by other techniques to constitute a constant and rather dense population of cells in the studied location. As an indication that we preserve the full complement of interstitial cells by our technique, these macrophage-like cells wer for the first time identified in material processed for scanning electron microscopy. The cells had characteristically irregular cell surfaces with short, veil-like, folded extensions intertwined between the other cells in the interstices. Our study establishes an improved correlation between results obtained by the application of scanning electron microscopy to dissected tissue and results from light and transmission electron microscopy of the intact tissue.

Animals↗
PubMed1991

Interstitial cells of Cajal in human small intestine. Ultrastructural identification and organization between the main smooth muscle layers.

Previous morphological and electrophysiological studies have supported the hypothesis that interstitial cells of Cajal have important regulatory (pacemaker) functions in the gut. In the current study, interstitial cells of Cajal associated with Auerbach's plexus in human small intestine were studied. Freshly resected intestine was examined by light and electron microscopy. The interstitial cells of Cajal resembled modified smooth muscle cells. They had caveolae and dense bodies, an incomplete basal lamina, a very well-developed smooth endoplasmic reticulum, and abundant intermediate (10 nm) filaments. Myosin filaments were not seen. Fibroblast-like cells were distinguished by their lack of caveolae and dense bodies, the relative scarcity of smooth cisternae and intermediate filaments, and the abundant granular endoplasmic reticulum. Interstitial cells of Cajal were arranged in networks of bundles containing processes of two to seven cells with fibroblastlike cells interspersed in the bundles. The bundles were innervated by nerve elements of Auerbach's plexus and extended into both layers of smooth muscle, between muscle cells, and into septa. The bundles were closely associated with elastin fibers. The organization shown in this study strongly supports the concept of interstitial cells of Cajal as important regulatory cells also in the human small intestine. The characteristic cytology and organization of interstitial cells of Cajal may provide a basis for future morphological, electrophysiological, and pathological studies of these cells in human small intestine.

Adult↗
PubMed1990

Prostaglandin H synthase immunoreactivity localized by immunoperoxidase technique (PAP) in the small intestine and kidney of rabbit and guinea-pig.

Prostaglandins and inhibitors of prostaglandin synthesis have striking regulatory effects on intestinal muscularis externa. We suggested earlier that a population of macrophage-like cells, located between the external muscle layers might release prostaglandins with a local effect on enveloping interstitial cells of Cajal, postulated pacemaker cells of the gut. To determine cellular production site(s) of prostaglandin we applied monoclonal antibodies against prostaglandin H synthase combined with the PAP technique to sections of rabbit and guinea-pig small intestine and kidney. In rabbit small intestine muscle cells in the circular muscle layer and in the muscularis mucosae were positive, longitudinal muscle negative. Vascular endothelial cells and serosal mesothelial cells were stained. In guinea-pig all muscle layers were unstained but endothelial and mesothelial cells were stained together with unidentified cells in the outermost submucosa. In rabbit kidney, positive staining of collecting ducts, interstitial cells, the parietal layer of Bowman's capsule and arterial endothelial cells was present. Furthermore, we found prostaglandin synthase antigenicity in the epithelial cells lining the loop of Henle, not described before. In guinea-pig medullary collecting ducts were stained and the papilla was lined by stained epithelial cells. The results show a species variation in the distribution of recognizable levels of prostaglandin H synthase. The impressive reaction in the mesothelium must be considered, when enzyme distribution is examined biochemically with fractionated tissue. Our findings do not support our hypothesis that macrophage-like cells are more potent sources of prostaglandins than smooth muscle cells.

Animals↗
PubMed1988

Macrophage-like cells in muscularis externa of mouse small intestine: immunohistochemical localization of F4/80, M1/70, and Ia-antigen.

Macrophage-like cells (MLC) in mouse small intestine are situated in the muscularis externa in the subserosal layer at the level of Auerbach's plexus, and at the level of the deep muscular plexus. By combined labelling with FITC-dextran and immunohistochemical techniques, the MLC were shown to express the macrophage markers F4/80 and M1/70.15. The MLC appeared to be constitutively IE-antigen-positive, but did not contain lysozyme. It is suggested that MLC, like Langerhans cells, belong to a specialized class of cells in the mononuclear phagocyte system.

Animals↗
PubMed1988

Selective double staining of interstitial cells of Cajal and macrophage-like cells in small intestine by an improved supravital methylene blue technique combined with FITC-dextran uptake.

By a brief exposure of small intestine (mouse, rat, guinea pig) to lysolecithin prior to vital methylene blue staining we were able to demonstrate in a selective way the complete network of interstitial cells of Cajal, located in the space between the longitudinal and circular muscle layers. A combination with fluorescence labeling (FITC-dextran uptake) of macrophage-like cells, allowed us to demonstrate 1) the complete, regular distribution of both cell populations along the entire small intestine, and 2) the constant, intimate associations between interstitial cells of Cajal and macrophage-like cells. In relation to current hypotheses concerning a role of interstitial cells of Cajal in motility regulation, our results call attention to the possible involvement of another cell type, the macrophage-like cell.

Animals↗
PubMed1985

Macrophage-like cells in the muscularis externa of mouse small intestine.

In muscularis externa of mouse small intestine, cells with ultrastructural features of macrophages were invariably observed in three layers: in the subserosal layer, between the circular and longitudinal muscle layers, and in association with the deep circular plexus. These macrophage-like cells (MLC) had a single indented nucleus, perinuclear Golgi complex, smooth and rough endoplasmic reticulum, many pits (coated and uncoated) in the plasma membrane, coated vesicles, light vesicles, and primary lysosomes, but rather few heterogeneous lysosomal vacuoles. MLC were partially enveloped by processes of interstitial cells of Cajal. FITC-dextran used in combined fluorescence stereo microscopy, fluorescence microscopy, and electron microscopy was employed as a tracer to study the endocytic qualities of the MLC. The mice were killed 5, 15, 30, and 60 min, 1 day, and 4 days after dextran administration. By fluorescence microscopy after 1 or 4 days MLC were observed as a constant cellular population with a strikingly regular distribution. By electron microscopy dextran-containing vacuoles were conspicuous after 1 h or more. MLC of the subserosal layer and between the circular and longitudinal muscle layers could be distinguished with respect to general appearance, pattern formation, and apparent dextran contents.

Animals↗
PubMed1982

Plexus muscularis profundus and associated interstitial cells. I. Light microscopical studies of mouse small intestine.

The zinc iodide/osmic acid (ZIO) method was used in a modification that selectively stained nerves and associated interstitial cells of Cajal (ICC) of muscularis externa. Due to its selectivity the method allowed a detailed stereoscopical analysis of whole mounts with respect to the topography and morphology of these elements. The method thus assisted and expanded our ultrastructural studies. The ZIO staining allowed a distinction of four morphologically different interstitial cell types (ICC-I-IV) confined to four compartments. The stained components were: (1) A rich plexus of highly ramified interstitial cells (ICC-II) in the subserous layer. (2) Auerbach's plexus with an associated extensive plexus of interstitial cells (ICC-I) in close contact with tertiary fasciculi. (3) Nerve fasciculi of the outer division of the circular muscle layer. These formed a nerve plexus in a well-defined plane in the outermost cell layers (plexus muscularis superficialis), with few fasciculi located internal to this plexus. A few bipolar interstitial cells (ICC-IV) were associated with nerve fasciculi of this region. (4) A nerve plexus located in the region between the two subdivisions of the circular muscle, plexus muscularis profundus (PMP). PMP was revealed throughout the small intestine as a continuous network of elongated, circularly oriented meshes. The pattern of connections between PMP and the other enteric plexuses was studied stereoscopically. Ganglion cells intrinsic to PMP occurred widely scattered. Interstitial cells associated with PMP (ICC-III) were arranged in a plexiform manner; their morphology and relations to nerves were investigated in great detail. A selective innervation of ICC-III via axons of PMP was strongly supported.

Animals↗
PubMed1982

Plexus muscularis profundus and associated interstitial cells. II. Ultrastructural studies of mouse small intestine.

The ultrastructure of plexus muscularis profundus (PMP) of the mouse small intestine was investigated subsequent to vascular perfusion with ruthenium red-containing and routine aldehyde fixatives. Four types of nerve terminals were revealed. Type I: numerous 500-A agranular vesicles and few 1,000-A granular vesicles. Type II: predominantly large (1,000-1,500 A), granular vesicles and fewer 500-A agranular vesicles. Type III: an abundance of mitochondria and many flattened vesicles (300 A X 700-1,300 A). Type IV was identified by abundant smooth cisternae 200 A in width. Types I-III formed close (200 A), synapse-like contacts to interstitial cells of Cajal (ICC-III). Presynaptic densities were frequent in type I endings. A direct innervation of muscle cells via PMP was only very occasionally suggested. ICC-III possessed a basal lamina and numerous caveolae associated with subsurface SER-cisternae. Mitochondria were very abundant in ICC-III-processes. ICC-III formed multiple, large gap junctions with outer circular-muscle cells and with other ICC-III. Also reflexive gap junctions were observed. Fibroblastlike cells (FLC) were distinguished by their prominent GER, the frequent presence of lipid droplets, and the lack of caveolae and a basal lamina. FLC never participated in synaptic arrangements or gap junctions. Macrophage-like cells were occasionally encountered. It is concluded that possible efferent and afferent nerve terminals in PMP may chiefly, if not exclusively, innervate ICC-III, the ultrastructure of which is compatible with efferent and/or afferent modulatory actions.

Animals↗