| 1. |
- Danielson, Patrik, et al.
(författare)
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Immunohistochemical and histochemical findings favoring the occurrence of autocrine/paracrine as well as nerve-related cholinergic effects in chronic painful patellar tendon tendinosis.
- 2006
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Ingår i: Microscopy research and technique (Print). - : Wiley. - 1059-910X .- 1097-0029. ; 69:10, s. 808-819
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Tidskriftsartikel (refereegranskat)abstract
- The pathogenesis of the pain in patellar tendon tendinosis ("jumper's knee") is unclear. We have recently presented new information about the sensory nervous system in the human patellar tendon, but there is very little information regarding the possible occurrence of a cholinergic system in this tendon. In the present study, specimens of pain-free normal tendons and chronically painful tendinosis tendons were examined by different immunohistochemical and histochemical methods. Antibodies against the M(2) receptor, choline acetyltransferase (ChAT), and vesicular acetylcholine transporter (VAChT) were applied, and staining for demonstration of activity of acetylcholinesterase (AChE) was also utilized. It was found that immunoreactions for the M(2) receptor could be detected intracellularly in both blood vessel cells and tenocytes, especially in tendinosis specimens. Furthermore, in the tendinosis specimens, some tenocytes were seen to exhibit immunoreaction for ChAT and VAChT. AChE reactions were seen in fine nerve fibers associated with small blood vessels in both the normal control tendons and the tendinosis tendons. The observations suggest that there is both a nerve related and a local cholinergic system in the human patellar tendon. As ChAT and VAChT immunoreactions were detected in tenocytes of tendinosis tendons, these cells might be a source of local acetylcholine (Ach) production. As both tenocytes and blood vessel cells were found to exhibit immunoreactions for the M(2) receptor, it is likely that both of these tissue cells may be influenced by ACh. Thus, in conclusion, there appears to be an upregulation of the cholinergic system, and an occurrence of autocrine/paracrine effects in this system, in the tendinosis patellar tendon.
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| 2. |
- Danielson, Patrik, et al.
(författare)
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In situ hybridization studies confirming recent findings of the existence of a local nonneuronal catecholamine production in human patellar tendinosis.
- 2007
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Ingår i: Microscopy research and technique (Print). - : Wiley. - 1059-910X .- 1097-0029. ; 70:10, s. 908-911
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
- We have in recent studies presented unexpected immunohistochemical evidence favoring the existence of a local production of catecholamines, and an occurrence of adrenergic receptors on the tendon cells (tenocytes), in the human patellar tendon. This was particularly noticed for tendons from patients suffering from tendinosis (chronic tendon pain), which has led us to propose an involvement of this autocrine/paracrine system in the development of tendinosis, especially since catecholamines have been reported to be modulators of tissue remodeling and pain processes. However, the findings concerning catecholamine production have so far only been noted at the level of protein detection, and for this reason, the aim of the present study was to confirm the previous immunohistochemical results by using in situ hybridization (ISH) technique. A ssDNA probe detecting human mRNA for the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH) was applied. The ISH results revealed that there were clear reactions indicating the existence of mRNA for TH in tenocytes of tendinosis specimens. It was generally noted that disfigured tenocytes were the ones with the most distinct reactions, while normally looking tenocytes hardly displayed any reactions at all. In conclusion, this study presents the first evidence at the mRNA level of the existence of a local nonneuronal production of catecholamines in human patellar tendon tissue. The findings add to recent observations of the occurrence of a local production in tendons of signal substances traditionally related to neurons. (c) 2007 Wiley-Liss, Inc.
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| 3. |
- Danielson, Patrik, et al.
(författare)
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Studies on the importance of sympathetic innervation, adrenergic receptors, and a possible local catecholamine production in the development of patellar tendinopathy (tendinosis) in man.
- 2007
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Ingår i: Microscopy research and technique (Print). - : Wiley. - 1059-910X .- 1097-0029. ; 70:4, s. 310-324
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Tidskriftsartikel (refereegranskat)abstract
- Changes in the patterns of production and in the effects of signal substances may be involved in the development of tendinosis, a chronic condition of pain in human tendons. There is no previous information concerning the patterns of sympathetic innervation in the human patellar tendon. In this study, biopsies of normal and tendinosis patellar tendons were investigated with immunohistochemical methods, including the use of antibodies against tyrosine hydroxylase (TH) and neuropeptide Y, and against alpha(1)-, alpha(2A)-, and beta(1)-adrenoreceptors. It was noticed that most of the sympathetic innervation was detected in the walls of the blood vessels entering the tendon through the paratendinous tissue, and that the tendon tissue proper of the normal and tendinosis tendons was very scarcely innervated. Immunoreactions for adrenergic receptors were noticed in nerve fascicles containing both sensory and sympathetic nerve fibers. High levels of these receptors were also detected in the blood vessel walls; alpha(1)-adrenoreceptor immunoreactions being clearly more pronounced in the tendinosis tendons than in the tendons of controls. Interestingly, immunoreactions for adrenergic receptors and TH were noted for the tendon cells (tenocytes), especially in tendinosis tendons. The findings give a morphological correlate for the occurrence of sympathetically mediated effects in the patellar tendon and autocrine/paracrine catecholamine mechanisms for the tenocytes, particularly, in tendinosis. The observation of adrenergic receptors on tenocytes is interesting, as stimulation of these receptors can lead to cell proliferation, degeneration, and apoptosis, events which are all known to occur in tendinosis. Furthermore, the results imply that a possible source of catecholamine production might be the tenocytes themselves. Microsc. Res. Tech., 2007. (c) 2007 Wiley-Liss, Inc.
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| 4. |
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| 5. |
- Lundberg, Simon, et al.
(författare)
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Natriuretic peptide expression in the heart of the TTR-ANP transgenic mouse-Comparison to the normal heart.
- 2005
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Ingår i: Microscopy Research & Technique. - : Wiley. - 1059-910X .- 1097-0029. ; 68:2, s. 97-106
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Tidskriftsartikel (refereegranskat)abstract
- The transgenic mice, TTR-ANP, carrying a fusion gene comprising the transthyretin promoter and the atrial natriuretic peptide (ANP) structural sequence, are known to have elevated ANP plasma levels as well as lowered blood pressure levels. On the other hand, it is not known whether these transgenic mice show changed natriuretic peptide (NP) expression patterns in the heart. Therefore, we examined the distribution of NPs in the myocardium and the conduction system of transgenic mice overexpressing ANP, as well as in wild-type littermates. The hearts were serially sectioned and processed for immunohistochemistry, with antisera against ANP and brain natriuretic peptide (BNP). Florescence microscopy was performed for qualitative analysis. Immunoreactivity for ANP and BNP was detected to a similar degree in the conduction system of both transgenic and wild-type mice. The nodal tissues never exhibited immunoreactions for ANP or BNP, whereas Purkinje fibers of the atrioventricular junctional tissue, bundle branches, and the peripheral Purkinje fiber network exhibited specific immunoreactivity. Atrial and ventricular myocytes of both transgenic and wild-type mice exhibited ANP and BNP to a similar extent. This is the first study examining the expression pattern of NPs in the cardiac conduction system of the mouse as well as the pattern of ANP and BNP expression in the conduction system of TTR-ANP transgenic mice and its wild-type siblings. The findings in this study suggest that ectopic ANP expression and release do not repress cardiac production of ANP.
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| 6. |
- Malmberg, Per, 1974, et al.
(författare)
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Imaging of lipids in human adipose tissue by cluster ion TOF-SIMS
- 2007
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Ingår i: Microsc Res Tech. - : Wiley. - 1059-910X .- 1097-0029. ; 70:9, s. 828-35
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Tidskriftsartikel (refereegranskat)abstract
- Biopsies of human subcutaneous adipose tissue were taken from healthy donors. Samples were high-pressure frozen, freeze-fractured, and freeze dried. Imaging mass spectrometry of samples was performed in a TOF-SIMS mass spectrometer equipped with a bismuth cluster ion source. Blood vessels, the connective tissue, and adipocytes can be seen in TOF-SIMS images. Blood vessels were found labeled by a high content of sodium ions and potassium ions in their lumen and phosphocholine signal in smooth muscle cells of the vessel wall. The connective tissue showed high signal levels of CN(-) fragments, derived from proteins and nucleic acids. Adipocytes showed high signal levels of phosphocholine and cholesterol ubiquitously in their membranes and diacylglycerols in some membrane sites. The central part of adipocytes showed high levels of triacylglycerols and fatty acids. These results are in accordance to those of biochemical studies; however, a precise spatial localization of lipids in adipocytes is demonstrated with MS imaging.
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| 7. |
- Nilsson, Harriet, et al.
(författare)
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A modified technique for the impregnation of lanthanum tracer to study the integrity of tight junctions on cells grown on a permeable substrate
- 2006
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Ingår i: Microscopy research and technique (Print). - : Wiley. - 1059-910X .- 1097-0029. ; 69:10, s. 776-783
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Tidskriftsartikel (refereegranskat)abstract
- Ionic lanthanum is commonly used to trace permeability pathways across epithelia and endothelia in biological electron microscopy. A method for obtaining a uniformly dense precipitate of lanthanum is described. The method, which is a modification of the technique described by Shaklai and Tavassoli (1977) was suitable for fixation of cell cultures grown on permeable filter inserts and was successfully applied to study opening of tight junctions by hypertonic solutions in the airway epithelial cell line 16HBE14o(-). The preparation method formed the basis for a semi-quantitative morphological determination in which the tight junctions were subdivided as "intact," "weakened," and "open." By using this modified technique, it could be demonstrated that opening of tight junctions in airway epithelial cells increased, with increasing osmolarity with electrolytes having a stronger effect than nonelectrolytes. A significant linear relationship was found between the osmolarity of the medium and the open state of the tight junctions (as determined by the semi-quantitative morphological technique) or the transepithelial electrical resistance.
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| 8. |
- Nygren, Håkan, 1952, et al.
(författare)
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Bioimaging TOF-SIMS: High resolution 3D imaging of single cells.
- 2007
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Ingår i: Microscopy research and technique. - : Wiley. - 1059-910X .- 1097-0029. ; 70:11, s. 969-74
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Tidskriftsartikel (refereegranskat)abstract
- The distribution of phosphocholine ions (m/z 184, m/z 86), sodium ions, and potassium ions in thyroid tumor cells was analyzed by imaging TOF-SIMS. Repeated sputtering with a C(60) (+) source and subsequent analysis with a Bi(3) (+) gun produced a series of 138 images that were stacked to make a 3D display of the chemistry of cells. Phosphocholine was seen in the plasma membrane (m/z 184) and intracellular membranes (m/z 86). The different fragmentation of the phospholipid probably reflects the chemical composition of membranes at these sites. High intensity of secondary ion signals of potassium was seen in membrane-encompassed cellular compartments. The data indicate that potassium ions are compartmentalized in thyroid tumor cells.
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| 9. |
- Nygren, Håkan, 1952, et al.
(författare)
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Imaging TOF-SIMS of rat kidney prepared by high-pressure freezing.
- 2005
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Ingår i: Microscopy research and technique. - : Wiley. - 1059-910X .- 1097-0029. ; 68:6, s. 329-34
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Tidskriftsartikel (refereegranskat)abstract
- Phosphocholine, potassium ions, and sodium ions were localized in rat kidney with imaging TOF-SIMS. Tissue preparation was performed with high-pressure freezing, freeze-fracturing and freeze-drying. The distribution of sodium ions was visualized by imaging the signal at m/z 23 of positively charged secondary ions, and the distribution of potassium ions was visualized by imaging the signal at m/z 39. Potassium was found localized within cells of the proximal tubulus epithelium and within cells of the glomeruli. High signals of sodium ions were seen in the interstitial tissue and also in epithelial cells of the collecting ducts and in glomeruli. The overlay image showed that the distribution of sodium ions and potassium ions were largely complementary with color mixing in glomeruli and in the interstitium surrounding proximal tubules. The ion distribution was further analyzed by correlation analysis. Phosphocholine-containing phospholipids were visualized by imaging the phosphocholine head group at m/z 184 of positively charged ions. The m/z 184 signal shows a ubiquitous distribution with a high intensity of phosphocholine in epithelial cells. Overlay image of m/z 184, m/z 39, and m/z 23 and multivariate analysis showed that the localization of high levels of phosphocholine colocalizes with high levels of potassium ions, as expected for an ion with intracellular localization.
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| 10. |
- Richter, Katrin, 1979, et al.
(författare)
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Localization of fatty acids with selective chain length by imaging time-of-flight secondary ion mass spectrometry.
- 2007
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Ingår i: Microscopy research and technique. - : Wiley. - 1059-910X .- 1097-0029. ; 70:7, s. 640-7
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Tidskriftsartikel (refereegranskat)abstract
- Localization of fatty acids in biological tissues was made by using TOF-SIMS (time-of-flight secondary ion mass spectrometry). Two cell-types with a specific fatty acid distribution are shown. In rat cerebellum, different distribution patterns of stearic acid (C18:0), palmitic acid (C16:0), and oleic acid (C18:1) were found. Stearic acid signals were observed accumulated in Purkinje cells with high intensities inside the cell, but not in the nucleus region. The signals colocalized with high intensity signals of the phosphocholine head group, indicating origin from phosphatidylcholine or sphingomyelin. In mouse intestine, high palmitic acid signals were found in the secretory crypt cells together with high levels of phosphorylinositol colocalized in the crypt region. Palmitic acid was also seen in the intestinal lumen that contains high amounts of mucine, which is known to be produced in the crypt cells. Linoleic acid signals (C18:2) were low in the crypt region and high in the villus region. Oleic acid signals were seen in the villi and stearic acid signals were ubiquitous with no specific localization in the intestine. We conclude that the results obtained by using imaging TOF-SIMS are consistent with known brain and intestine biochemistry and that the localization of fatty acids is specific in differentiated cells.
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