Staining of constitutive heterochromatin in mammalian chromosomes with a new fluorochrome

Hilwig, I.; Gropp, A.

doi:10.1016/0014-4827(72)90527-7

Cited by 321 publications

(67 citation statements)

References 13 publications

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“…For nuclear staining of parenchymal tissue, the fluorescent dye bis-benzamide H33342 (2 µmol/100 g body weight; Sigma) and ultraviolet epi-illumination (330-390 nm/430 nm) were used. 14 This dye binds to DNAs in living cells 15,16 and stains within the liver primarily the nuclei of hepatocytes but not Kupffer cells or endothelial cells. 14 Quantitative analysis was performed off-line by a computerassisted image analysis system (CapImage; Zeintl, Heidelberg, Germany).…”

Section: Methodsmentioning

confidence: 99%

An intravital fluorescence microscopic study of hepatic microvascular and cellular derangements in developing cirrhosis in rats

1998

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Quantitative data defining the relationship between the hepatic microcirculation and the development of liver pathological changes could provide a basis for a better understanding of fibrogenic processes, such as cirrhosis. Therefore, we established the technique of intravital fluorescence microscopy and computer-assisted microcirculation analysis systems in developing cirrhosis in rats with the aim of quantitatively assessing the association of hepatic microvascular morphology with its disordered acinar architecture, and nonparenchymal cell transformation with collagen deposition, parenchymal cell loss, and liver dysfunction. In animals chronically exposed to carbon tetrachloride The pathological process of developing cirrhosis represents an almost inconceivably complex network of interacting cells, fibrogenic mediators, and extracellular matrix molecules (ECM). 1 Dramatic improvements in cell and molecular biological techniques now permit a detailed analysis of the composition, structure, immunology, and histological localization of the individual components of extracellular matrix. Dissection of the cellular and molecular pathobiology of ECM deposition includes the identification of the cellular sources of specific ECM components as well as the definition of humoral factors and regulatory loops underlying both the amplification and degradation pathways of ECM molecules during fibrogenesis. 2 Beside these molecular aspects of fibrogenesis, vascular modifications during the development of cirrhosis have fully been recognized and interpreted in that microvascular changes in the liver are not the mere consequence of fibrosis and nodule formation, but dominate the entire development of the pathological condition. 3 Microvascular corrosion casts observed by scanning electron microscopy have proved useful in demonstrating microvascular morphology in hepatic cirrhosis [4][5][6][7][8] ; however, these techniques are neither suited for the investigation of dynamic alterations in the microvascular function nor for the direct assessment of the close interrelation between the hepatic microcirculation and liver structure and function. Combining several applications of intravital fluorescence microscopy in a rat model of developing cirrhosis, the present study provides the first in vivo appreciation of the association of microvascular, cellular, and functional aspects during the process of hepatic organ remodeling. MATERIALS AND METHODSAnimal Model.

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Section: Methodsmentioning

confidence: 99%

An intravital fluorescence microscopic study of hepatic microvascular and cellular derangements in developing cirrhosis in rats

1998

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“…Control studies in which normal rabbit serum was substituted for either one or both of the primary antisera verified the specificity of this procedure. Nuclei were stained with Hoechst 33258 (.04 pg/ml for 10 min) following immunohistochemistry (Hilwig and Gropp, 1972).…”

Section: Immunohistochemical Methodsmentioning

confidence: 99%

Immunohistochemical studies indicate multiple enteroendocrine cell differentiation pathways in the mouse proximal small intestine

Aiken

Kisslinger

Roth

1994

Developmental Dynamics

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The enteroendocrine cell system of the mammalian gastrointestinal tract is comprised of at least 16 different subpopulations. Each subpopulation shows a characteristic distribution along both the crypt-villus and cephalocaudal axes. In both the small intestine and colon of adult mice, multilabel immunohistochemistry has demonstrated that two or more neuroendocrine products can be coexpressed in various combinations in single cells along the crypt-villus axis, suggesting that enteroendocrine phenotypes may be actively regulated. Using bromodeoxyuridine (BrdU) incorporation and multilabel immunohistochcmistry, we have previously demonstrated an enteroendocrine cell differentiation pathway consisting of two subpopulations of cells in the mouse proximal small intestinmne involving the sequential expression of substance P, serotonin, and secretin in cells migrating out of the crypts into the villi, and a second involving the expression of substance P and serotonin in cells which remain in the crypts. In this report, we use double label immunohistochemistry and BrdU incorporation to define the temporal and spatial interrelationships between gastrin, cholecystokinin (CCK), glucagon-like peptide-1 (GLP-I), and gastric inhibitory peptide (GIP) immunoreactive cells in the mouse proximal small intestine. The expression of these products was compared with that of substance P, serotonin, and secretin. Minimal overlap of expression was found in cells immunoreactive for substance P or serotonin with gastrin, CCK, GLP-1, or GIP; however, secretin was found colocalized in villus-associated gastrin, CCK, and GLP-1 containing cells. We demonstrate that, similar to the bidirectionally migrating substance P and serotonin expressing cells, gastrin, CCK, GLP-1, and secretin are expressed in upwardly migrating cells, and gastrin, CCK, and GLP-1 are expressed in downwardly migrating cells that fail to express secretin. GIP containing cells only rarely coexpressed any of the products examined, but were found both in the villi and the crypts, suggesting both upwardly and downwardly migrating populations. These findings demonstrate several novel enteroendocrine cell differentiation pathways. In addition, the expression of secretin 0 1994 WILEY-LISS, INC.in the villi, but not in the crypts, by two otherwise distinct differentiation pathways, and the lack of secretin expression by villus-associated GIP expressing cells, suggests that local factors present in the crypts and/or on the villi are necessary, but not sufficient, for secretin expression.

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“…This was followed by application of the second primary antiserum and then a fluorescent labeled secondary antiserum (Jackson ImmunoResearch, West Grove, PA) or detection by the peroxidase anti-peroxidasei naphthol basic dye method (Mauro et al, 1985;Hermiston et al, 1992). Nuclei were stained with Hoechst 33258 (.04 p.g/ml for 10 min) following immunohistochemistry (Hilwig and Gropp, 1972).…”

Section: Immunohistochemical Methodsmentioning

confidence: 99%

Temporal differentiation and migration of substance P, serotonin, and secretin immunoreactive enteroendocrine cells in the mouse proximal small intestine

Aiken

Roth

1992

Developmental Dynamics

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Precise spatial interrelationships exist between substance P, serotonin, and secretin containing enteroendocrine cells in the gastrointestinal tract of mice. In the proximal small intestine these products are coexpressed in various combinations in single enteroendocrine cells along the crypt to villus axis in a pattern that suggests the sequential expression of substance P, serotonin, and secretin. In this report we use bromodeoxyuridine (BrdU) and multilabeling immunohistochemistry to define the temporal and spatial interrelationships between substance P, serotonin, and secretin immunoreactive cells in the mouse proximal small intestine. Our findings demonstrate the sequential expression of substance P, serotonin, and secretin in a population of upwardly migrating enteroendocrine cells and, furthermore, identify a population of crypt associated cells coexpressing substance P and serotonin that fails to traverse this pathway. The lack of secretin immunoreactive cells in the crypts suggests that local factors present in the crypts and/or on villi regulate secretin expression. The combined use of BrdU and multilabeling immunohistochemistry provides a method for defining enteroendocrine cell differentiation pathways throughout the gastrointestinal tract. 0 1992 Wiley-Liss, Inc.

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Staining of constitutive heterochromatin in mammalian chromosomes with a new fluorochrome

Cited by 321 publications

References 13 publications

An intravital fluorescence microscopic study of hepatic microvascular and cellular derangements in developing cirrhosis in rats

An intravital fluorescence microscopic study of hepatic microvascular and cellular derangements in developing cirrhosis in rats

Immunohistochemical studies indicate multiple enteroendocrine cell differentiation pathways in the mouse proximal small intestine

Temporal differentiation and migration of substance P, serotonin, and secretin immunoreactive enteroendocrine cells in the mouse proximal small intestine

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