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  • 1
    Electronic Resource
    Electronic Resource
    Direct evidence for the binding of rat liver DPP IV to collagen in vitro
    Amsterdam : Elsevier
    Experimental Cell Research 178 (1988), S. 64-72 
    Details
    ISSN: 0014-4827
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/0014-4827(88)90378-3
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  • 2
    Electronic Resource
    Electronic Resource
    Topographische und histologische Untersuchungen am Reizleitungssystem von Goldhamster, Mäusen und Spitzmäusen (1972)
    Springer
    Anatomy and embryology 137 (1972), S. 114-124 
    Details
    ISSN: 1432-0568
    Keywords: Heart ; Impulse conducting system ; Golden hamsters ; Mice ; Shrews
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Das Reizleitungssystem (RLS) von Goldhamster, Maus und Ratte wird färberisch-lichtmikroskopisch untersucht. 1. Bei allen species besteht das RLS aus Sinusknoten und Atrioventricularsystem (AV-Knoten,-Bündel, linker und rechter Kammerschenkel mit Endaufzweigungen), die sich prinzipiell nicht von denen anderer Säuger unterscheiden. 2. Histologisch ist das RLS sämtlicher Arten außerordentlich muskelfaserreich bzw. bindegewebsarm und ähnelt vor allem bei Goldhamster und Maus stark der Arbeitsmuskulatur. — Eine Bindegewebsscheide im Gebiet des AV-Systems ist besonders deutlich bei den Spitzmäusen ausgeprägt. — Eine spezielle Gefäßversorgung kommt nur im Sinus-und AV-Knoten von Goldhamster und Mäusen vor. 3. Regionale Unterschiede finden sich bevorzugt innerhalb des AV-Systems von Gold-hamster und Maus, weniger bei Spitzmäusen. 4. Artspezifische Differenzen existieren zwischen dem RLS von Goldhamstern und Mäusen einerseits und dem der Spitzmäuse andererseits. 5. Verglichen mit dem RLS größerer Säuger ist das von Goldhamster, Maus und Spitzmaus wesentlich kompakter und dem Myokard abschnittsweise weitgehend gleich gebaut. Dies dürfte zusammen mit der geringen Größe der Hauptgrund für die Schwierigkeiten sein, das RLS bei den genannten Arten sicher zu lokalisieren.
    Notes: Summary The impusle conducting system (ICS) of golden hamsters, mice and shrews was investigated by means of light microscopy. 1. The ICS of all species mentioned consists of a sinatrial node and an atrioventricular system (AV node, bundle, left and right limb, terminal branchings) in principal not differing from those in other mammals. 2. Histologically the ICS of golden hamsters, mice and shres is conspicuously rich in mucsle fibres respectively poor in connective tissue and rather similar to the myocardium, especially in the heart of golden hamsters and mice.—In the region of the AV system of shrews a connective tissue sheath is well developed.—A special blood supply can only be observed in the sinatrial and AV node of golden hamsters and mice. 3. Regional differences preponderantly exist within the AV system of golden hamsters and mice; in shrews its structure seems to be more homogenous. 4. Species-dependant differences can be found between the ICS of golden hamsters and mice on the one and that of shrews on the other side. 5. In comparison with that of bigger mammals the ICS of golden hamsters, mice and shrews is characterized by compact arrangement of its muscle fibres and striking histological similarity with the working myocardium which might be—together with the small size—the main reason for the difficulties to localize the ICS in the heart of these species exactly.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00523533
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  • 3
    Electronic Resource
    Electronic Resource
    The basement membrane of the persisting maternal blood vessels in the placenta of Callithrix jacchus (1987)
    Springer
    Anatomy and embryology 176 (1987), S. 87-97 
    Details
    ISSN: 1432-0568
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Formation and morphology of the thickened basement membrane-like layer around the persisting maternal vessels of the Callithrix jacchus placenta were investigated from day 45 until term (day 142) using light, electron and immunofluorescence microscopy. Thickening occurs with the establishment of contacts between the vessels and the syncytiotrophoblast (day 48). Final thickness is reached at about day 100. The course of the vessels shows wide gaps where the maternal blood flows freely into the intertrabecular spaces. As revealed by electron microscopy, the extracellular sheath around the maternal vessels consists of an inner subendothelial basement membrane (3–6 μm) and an outer fibril-containing layer (2–4 μm). Cell debris is seen between the two layers and in the basement membrane. Plaques of granular and fine-filamentous material are incorporated into the fibril-containing layer. The synthesis of the basement membrane material is localized in the endothelial cells. Immunofluorescence microscopy reveals collagen types IV and V, laminin and heparan sulfate proteoglycan (BM-1) in the sheath around the persisting vessels. Fibronectin occurs only in certain areas or in the form of dots. Collagen types I and III are not seen in the region of the vascular wall. It can, therefore, be assumed that the subendothelial layer represents a genuine basement membrane; the fibrils consist of collagen type V and the plaques contain fibronectin. The existence of the thick perivascular sheath is attributed to the persistence and stability of the maternal vessels.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00309756
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  • 4
    Electronic Resource
    Electronic Resource
    Placental toxicity in rats after administration of synthetic glucocorticoids (1989)
    Springer
    Anatomy and embryology 180 (1989), S. 121-130 
    Details
    ISSN: 1432-0568
    Keywords: Pregnant rats ; Glucocorticoids ; Glycogen ; Proteases ; von Willebrand factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Administration of the synthetic glucocorticoids dexamethasone and triamcinolone to pregnant rats between gestational day (GD) 16 and 20 caused dose-dependent placental lesions on GD 21 and 22 which were detected by morphological, histochemical and immunohistochemical means. Maternal blood spaces, trophoblast layer and fetal blood vessels were altered primarily in the centre of the placentral labyrinth. Less severe changes were found in the junctional zone, chorionic plate and intraplacental yolk sac. On GD 21, low doses increased the amount of glycogen, while high doses induced a loss of glycogen. γ-glutamyl transpeptidase activity was increased in the spongiotrophoblast and the labyrinthic trophoblast and dipeptidyl peptidase IV activity in fetal capillary endothelium, whereas α-glutamyl aminopeptidase and microsomal alanyl aminopeptidase were not affected. Additionally, in the fetal capillary endothelium an increase of immunoreactivity for the von Willebrand factor occurred. These data suggest that synthetic glucocorticoids affect placental tissues at different and rather specific levels, which may in turn disturb placental function and contribute to fetal maldevelopment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00309763
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  • 5
    Electronic Resource
    Electronic Resource
    Increased activity of dipeptidyl peptidase IV in serum of hepatoma-bearing rats coincides with the loss of the enzyme from the hepatoma plasma membrane (1986)
    Springer
    Cellular and molecular life sciences 42 (1986), S. 826-828 
    Details
    ISSN: 1420-9071
    Keywords: Dipeptidyl peptidase IV ; hepatoma rats ; cancer markers ; immunofluorescence ; cytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The specific activity of dipeptidyl peptidase IV (DPPIV E.C. 3.4.14.-) in the plasma membrane of Morris hepatoma 9121 or hepatoma 7777 was 3.5% and 2.9%, respectively, of that in the plasma membrane of rat liver. The enzyme activity in the serum of hepatoma-bearing rats was 141% (hepatoma 91219) and 162% (hepatoma 7777) of the normal value. cytochemical investigation showed that the DPP IV activity was almost completely absent from the hepatoma cell plasma membrane and was not sequestered within these cells. Indirect immunofluorescence staining with a polyclonal antibody directed against DPP IV indicated that the loss of activity was due to the absence of DPP IV molecules in the plasma membrane. The possibility that the enzyme is transferred from the membrane into the serum as a result of structural alterations is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01941540
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  • 6
    Electronic Resource
    Electronic Resource
    Histochemische, fluoreszenzmikroskopische und experimentelle Untersuchungen am Reizleitungssystem von Goldhamster, Maus und Ratte (1971)
    Springer
    Histochemistry and cell biology 26 (1971), S. 44-60 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Im Reizleitungssystem (RLS) von Goldhamster, Maus und Ratte sind nicht nur die oxidativen sondern auch die glykolytischen Enzymaktivitäten geringer als in der Arbeitsmuskulatur. Innerhalb der Reizleitungsmuskulatur überwiegt die Glykolyse. Demgegenüber besitzen Glucose-6-phosphat-Dehydrogenase und lysosomale Enzyme im RLS höhere Aktivitäten als im Myokard. Die Lactat-Dehydrogenase-Isoenzyme verhalten sich in beiden Muskelgeweben weitgehend gleich. Entsprechend dem reduzierten glykolytischen und oxidativen Stoffwechsel ist die Kapillardichte im RLS gering. Die Aktivitäten der Enzyme des Glykogenstoffwechsels laufen in RLS und Myokard in etwa dem Glykogengehalt parallel. Klare artspezifische und regionale Stoffwechselunterschiede fehlen innerhalb der Reizleitungsmuskulatur. Durch hohe cholinerge Nervenfaserdichte zeichnen sich alle Teile des RLS von Goldhamster und Maus aus. Eine dichte adrenerge Innervation ist nur beim Goldhamster im gesamten RLS anzutreffen. Im Mäuseherzen bevorzugen die adrenergen Fasern den Sinusknoten. Im AV-System von Ratten nimmt das Glykogen nach Hypoxie, Katecholamingaben und Hunger ab. Beim Goldhamster tritt überzeugender Glykogenverlust im AV-System nur nach Sauerstoffmangel und Katecholaminapplikation ein.
    Notes: Summary In the impulse conducting system (ICS) of golden hamsters, mice and rats the activities of glycolytic as well as of oxidative enzymes are lower than in the myocardium. However, in the ICS the glycolytic pathway is relatively more active in comparison with the oxidative metabolism. On the contrary, glucose-6-phosphate dehydrogenase and the lysosomal enzymes activities are higher in the ICS than in the ordinary cardiac muscle. The lactate dehydrogenase isoenzyme pattern is nearly the same in both heart muscle tissues. In agreement with the lowered glycolytic and oxidative metabolism a reduced capillarisation is found in the ICS. The activities of enzymes involved in glycogen metabolism are related to the amount of glycogen in the impulse conducting tissue and working muscle. Within the ICS clear speciesdependent and regional metabolic differences have not been observed. A dense cholinergic innervation occurs in all parts of the ICS of golden hamsters and mice. An overall rich adrenergic innervation is typical only for the ICS of golden hamsters. In the mouse heart, however, the adrenergic nerve fibers prefer the sinatrial node. In the AV system of rats glycogen decreases following hypoxia, catecholamine application and hunger. In the same ICS-region of golden hamsters a marked loss of glycogen can only be produced by hypoxia and treatment with catecholamines.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00307783
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  • 7
    Electronic Resource
    Electronic Resource
    Verwendung der Gefriertrocknung nach Lowry in der Histochemie (1972)
    Springer
    Histochemistry and cell biology 29 (1972), S. 185-188 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Ausgehend von der mikrochemischen Gefriertrocknung von Kryostatschnitten wird ein Verfahren beschrieben, mit dem parallel mikrochemische und histochemische Enzymnachweise durchgeführt werden können. Anhand praktischer Beispiele werden die Vorteile dieser Methode für histochemische Untersuchungen geschildert.
    Notes: Summary Basing on the microchemical freeze-drying of cryostat sections a simple procedure was developed which allows the parallel demonstration of enzymes with microchemical and histochemical methods. The advantages of this technique for histochemical investigations are described and some practical examples for its use are given.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00277287
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  • 8
    Electronic Resource
    Electronic Resource
    Über den Histochemischen Nachweis der β-Glucuronidase, α-Mannosidase und α-Galactosidase mit 1-Naphthylglykosiden (1973)
    Springer
    Histochemistry and cell biology 36 (1973), S. 367-381 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Mit simultanen Azokupplungsverfahren und 1-Naphthylglykosiden als Substraten werden Verteilung und Aktivität von β-Glucuronidase, α-Mannosidase und α-Galactosidase bei Ratte, Maus und Meerschweinchen untersucht. Für die β-Glucuronidase besteht das Inkubationsmedium aus 5–10 mg 1-Naphthylβ-glucuronid (gelöst in 0.4 ml NN-Dimethylformamid) und 0.6 ml 2% Hexazonium-p-rosanilin in 9 ml 0.2 M Acetat-Puffer, pH 5; für die α-Mannosidase und α-Galactosidase aus der gleichen Menge 1-Naphthyl-α-mannosid bzw. -α-galactosid und p-Rosanilin in 9 ml 0.1 M CitratCitronensäure-Phosphatoder Acetat-Puffer, pH 5 bzw. 5.2. Die Spezifität der Nachweisreaktionen sichern qualitative und quantitative Hemmversuche mit verschiedenen 1–4-Lactonen und Galactose ab. Die β-Glucuronidase kann bei der Ratte vor allem intralysosomal nachgewiesen werden, z.B. in Niere, Nebenhoden, Uterus, Samenblase, Darm und Respirationstrakt; Mäuseund Meerschweinchengewebe setzen 1-Naphthyl-β-glucuronid langsamer um. Für die α-Mannosidase läßt sich in zahlreichen Organen auch histochemisch die lysosomale Lokalisation des Enzyms beweisen, wobei die Aktivität in Urogenitalsystem, Darm und Speicheldrüsen besonders hoch ist, und in der Mäuseniere geschlechtsspezifische Unterschiede vorkommen. Erstmalig wird die intralysosomale Lokalisation der α-Galactosidase gezeigt, die ubiquitär in teilweise hoher Aktivität anzutreffen ist. 6-Br-2-Naphthyl-α-galactosid eignet sich in Verbindung mit Fast Blue B nicht zur intralysosomalen Lokalisation der α-Galaotosidase. Fluorometrische Messungen aller 3 Glykosidasen mit dem entsprechenden 1-Naphthylglykosid ergeben nach Fixation in Formol oder Glutaraldehyd Hemmraten zwischen 90 und 98 %; anschließendes Waschen in Zuckerlösung verdoppelt oder verdreifacht die Restaktivität.
    Notes: Summary By means of simultaneous azo coupling using 1-naphthyl glycosides as substrates the distribution and activity of β-glucuronidase, α-mannosidase and α-galactosidase have been investigated in rats, mice and guinea-pigs. For β-glucuronidase the incubation medium consists of 5–10 mg 1-naphthyl-β-glucuronide (dissolved in 0.4 ml NN-dimethyl formamide) and 0.6 ml 2% hexazonium-p-rosaniline in 9 ml 0.2 M acetate buffer, pH 5; for α-mannosidase and α-galactosidase of the same quantities of 1-naphthyl-α-mannoside and α-galactoside respectively and p-rosaniline in 9 ml 0.1 M citrate, citric acid-phosphate or acetate buffer, pH 5. Qualitative and quantitative inhibition tests using various 1–4 lactones and galactose prove the reaction specifity of the methods presented here. β-Glucuronidase can be detected especially in lysosomes of rat organs, e.g. kidney, epididymis, uterus, vesicular gland, intestine and respiratory tract; tissues from mice and guineapigs exhibit a slower splitting rate for 1-naphthyl glucuronide. As to α-mannosidase its lysosomal localization becomes apparent in many organs also by means of histochemistry. The urogenital system, intestine and the salivary glands belong to the structures with the highest amount of α-mannosidase, and in the mouse kidney sex differences occur. For the first time α-galactosidase can be demonstrated unequivocally in the lysosomes of rat, mouse and guineapig tissues in which this enzyme displays a high overall activity. 6-Br-2-Naphthyl-α-galactoside and Fast Blue B for postcoupling are not able to detect the lysosomal localization of α-galactosidase. Fluorometric measurements of these 3 glycosidases by means of the corresponding 1-naphthyl glycoside reveal inhibition rates between 90 and 98% following fixation in formol or glutaraldehyde. Washing in sugar solution raises enzyme activity two or three times.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00305715
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  • 9
    Electronic Resource
    Electronic Resource
    Untersuchung der N-Acetyl-β-glucosaminidase mit 1-Naphthyl-N-Acetyl-β-glucosaminid (1973)
    Springer
    Histochemistry and cell biology 37 (1973), S. 169-185 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Mit 1-Naphthyl-N-acetyl-β-glucosaminid als Substrat wird die N-Acetyl-β-glucosaminidase (N-A-Gase) histochemisch (Simultankupplung von 1-Naphthol an Hexazonium-p-rosanilin) und mikrochemisch (fluorometrische Messung von 1-Naphthol) in Ratten-, Mäuseund Meerschweinchenorganen untersucht. Das histochemische Inkubationsmedium enthält 5–12 mg 1-Naphthyl-N-acetyl-β-glucosaminid (gelöst in NN-Dimethylformamid) und 0,8–1 ml 2% Hexazonium-p-rosanilin in 9 ml 0,1 M Citrat-Puffer, pH 5; das mikrochemische 2 mg/ml dieses Substrats (6 mM) im gleichen Puffer, pH 4,5. Zum Nachweis in situ empfiehlt sich wegen der geringeren Hemmrate (32%) Formol-Fixation, in hochaktiven Geweben auch Glutaraldehyd (54% Inhibition im proximalen Konvolut der Rattenniere). Die histochemische Gesamtaktivität der N-A-Gase ist besser an frischen Schnitten mit semipermeablen Membranen zu erfassen. Nach Fixation in Glutaraldehyd kann das Enzym vor allem bei Ratten in den Lysosomen zahlreicher Organe nachgewiesen werden (u.a. Niere, Nebenhoden, Bronchien, Darm, Uterus und Samenblase), wobei artspezifische Unterschiede bestehen: Über die höchste N-A-Gase-Aktivität verfügen Nebenniere, Nebenhoden und Milz von Meerschweinchen. Weniger Enzym enthalten diese Organe bei Ratten und Mäusen; speziell die Nebenniere ist weitgehend N-A-Gase-frei. Am kräftigsten reagiert die Niere bei Ratten. Geschlechtsspezifische Differenzen sind hier nur mikrochemisch faßbar, in der Niere der Maus aber schon histochemisch. — Reich an N-A-Gase sind Speicheldrüsen, Harnblase und Colon sowie andere Organe mit mucopolysaccharid-haltigen Strukturen. Die mit der Naphthol-AS und 1-Naphthyl-Methode für die N-A-Gase erhaltenen Aktivitätsund Verteilungsmuster entsprechen sich. Die Qualität der intrazellulären Enzymlokalisation mit 1-Naphthyl-N-acetyl-β-glucosaminid als Substrat kommt den Möglichkeiten des Naphthol-AS-Verfahrens in vielen Organen nahe oder ist ihm gleichwertig.
    Notes: Summary Using 1-naphthyl-N-acetyl-β-gIucosaminide as substrate N-acetyl-β-glucosaminidase (N-A-Gase) has been investigated histochemically (simultaneous coupling of 1-naphthol and hexazonium p-rosaniline) and microchemically (fluorometric measurement of 1-naphthol) in various organs of rats, mice, and guinea-pigs. The histochemical incubation medium consists of 5–12 mg 1-naphthyl-N-acetyl-β-glucosaminide (dissolved in NN-dimethyl formamide) and 0.8–1 ml hexazotized p-rosaniline in 9 ml 0.1 M citrate buffer, pH 5; for microchemical purposes 2 mg/ml of this substrate (6 mM) in the same buffer, pH 4.5 were used. For the in situ demonstration of N-A-Gase formol fixation is recommended because of its lower inhibition rate (32%); in highly active tissues glutaraldehyde is also suitable (54% inhibition in the proximal convoluted tubules of the rat kidney). The total activity of N-A-Gase can better be detected in fresh frozen sections in connection with semipermeable membranes. Following fixation in glutaraldehyde the enzyme occurs in the lysosomes of many organs, e.g. kidney, epididymis, bronchi, adrenal gland, intestine, uterus, and vesicular gland, especially in rats. Furthermore spezies-dependent differences exist: the suprarenal gland, epididymis, and spleen of guinea-pigs display the highest amount of N-A-Gase. In rats and mice the enzyme activity of these organs is lower; the adrenal cortex is nearly free of N-A-Gase. — The kidney reacts intensely in rats, the sex differences of which can only be detected by means of microchemistry. In the mouse kidney they are more pronounced. Therefore the histochemical N-A-Gase assay reveals them, too. — Organs containing considerable quantities of mucopolysaccharides, e.g. the submandibular gland, urinary bladder, and colon are also rich in N-A-Gase. The activity and distribution pattern of N-A-Gase obtained with the naphthol AS and 1-naphthyl technique are completely in correspondance with one another. In some cases the quality of the intracellular enzyme localization using naphthol AS-BI N-acetyl-β-glucosaminide as substrate surpasses the possibilities of the 1-naphthyl derivate, in others the latter one enables identical results.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00305588
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  • 10
    Electronic Resource
    Electronic Resource
    Azoindoxylverfahren zum Hydrolasennachweis (1978)
    Springer
    Histochemistry and cell biology 57 (1978), S. 323-342 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung An frischen, gefriergetrockneten und Schnitten von aldehyd-fixierten Rattengeweben werden 13 Diazoniumsalze als Simultankuppler zum Nachweis saurer, neutraler und alkalischer Hydrolasen mit Azoindoxylverfahren geprüft. Hexazotiertes Neufuchsin und/oder Fast Blue B sind die Diazoniumsalze der Wahl zur Lokalisation von saurer β-Galactosidase, Neuraminidase, β-N-Acetylglucosaminidase, saurer Phosphatase und unspezifischer Esterase gefolgt von Hexazonium-p-rosanilin. Fast Blue VB, BB und RR sowie Fast Violet B eignen sich zur Untersuchung von Lactase und alkalischer Phosphatase; Fast Garnet GBC kann zur Lokalisation von saurer β-Galactosidase, Glucosaminidase und Lactase, Fast Red B, RC, RL und TR sowie Black K nur für Lactase-Studien verwandt werden. Durchschnittlich reichen 0,01–0,02 ml instabiles Diazoniumsalz und 0,3–1 mg stabiles Diazoniumsalz/ml zur korrekten Lokalisation dieser Hydrolasen aus. Im einzelnen hängt die Kupplerkonzentration von der Enzymaktivität und vom untersuchten Organ ab. Gefriergetrocknete Kryostatschnitte liefern unabhängig vom Kuppler die besten Resultate bei Untersuchung von Lactase und alkalischer Phosphatase; Schnitte von form- oder glutaraldehyd-fixierten Organen sind beim Nachweis der restlichen Hydrolasen überlegen. Eine Ausnahme macht die Untersuchung der sauren β-Galactosidase und Glucosaminidase mit Fast Garnet GBC; dann werden die besten Ergebnisse nach Gefriertrocknung erzielt. Frische Kryostatschnitte sind zur Darstellung der Lactase mit hexazotiertem Neufuchsin oder p-Rosanilin und der alkalischen Phosphatase mit Fast Blue VB und BB sowie Violet B geeignet; die Gesamtaktivität der sauren, neutralen und alkalischen Hydrolasen kann mit semipermeablen Membranen und den stabilen sowie instabilen Diazoniumsalzen der Wahl untersucht werden. Ausreichende Osmierung der Azoindoxylfarbstoffe ist nur möglich, wenn Hexazonium-p-rosanilin als Kupplungsreagens benutzt wird; ohne Vorbehandlung extrahieren Äthanol, Isopropanol und Xylol alle Azoindoxyle. 7 Inkubationsmedien werden zum lichtmikroskopisch-histochemischen Nachweis von Glykosidasen, Esterasen und Phosphatasen mit Azoindoxylmethoden angegeben und typische Anwendungsbeispiele genannt. Zur Gefriertrocknung von Kryostatschnitten mit dem Edwards-Pearse Gewebetrockner EPD 3 wird ein modifiziertes Verfahren beschrieben.
    Notes: Summary Using fresh frozen, freeze-dried or cryostate sections from aldehyde fixed rat tissues 13 diazonium salts were tested as simultaneous coupling reagents for the localization of acid, neutral and alkaline hydrolases with azo indoxyl methods. Hexazotized new fuchsine and/or Fast blue B are the diazonium salts of choice for the demonstration of acid β-galactosidase, neuraminidase, β-N-acetylglucosaminidase, acid phosphatase, and non-specific esterase followed by hexazotized p-rosaniline. Fast blue VB, BB and RR and Fast violet B are recommended for the investigation of alkaline phosphatase and lactase, Fast garnet GBC for acid β-galactosidase, glucosaminidase and lactase. Fast red B, RC, RL and TR and Fast black K can only be employed for lactase studies. The exact concentration of the coupling reagent depends on the activity of the enzyme and the organ investigated. On the average 0.01–0.02 ml unstable diazonium salt/ml and 0.3–1 mg stable diazonium salt/ml are sufficient for the correct localization of these hydrolases. Freeze-dried cryostat sections yield the best results in the demonstration of lactase and alkaline phosphatase independent on the coupling reagent used. Sections from formaldehyde or glutaraldehyde fixed organs are superior for the localization of the other hydrolases; an exception is the investigation of acid β-galactosidase and glucosaminidase with Fast garnet GBC. Then, excellent results are obtained also with freeze-dried material. Fresh frozen sections are suitable for the localization of lactase with hexazotized new fuchsine or p-rosaniline and of alkaline phosphatase with Fast blue VB and BB or violet B. The total activity of acid, neutral and alkaline hydrolases can be investigated using semipermeable membranes in combination with all unstable and stable diazonium salts of choice. Reliable osmification of the azoindoxyl dye is only possible if hexazotized p-rosaniline is employed for coupling; without further posttreatment all azoindoxyl dyes are extracted by ethanol, isopropanol or xylol. 7 incubation media are given for the demonstration of hydrolases with azoindoxyl methods at the level of light microscopy for routine studies and typical examples for the application of these methods are presented. A modified procedure is described for the freeze-drying of cryostat sections with the Edwards-Pearse tissue dryer EPD 3.
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    URL: http://dx.doi.org/10.1007/BF00492668
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