In Situ Hybridization and Double Immunohistochemistry for the Detection of VEGF-A mRNA and CD34/Collagen IV Proteins in Renal Transplant Biopsies

2017 ◽  
pp. 131-143
Author(s):  
Dejan Dobi ◽  
Zoltan G. Laszik
Keyword(s):  
In Situ Hybridization ◽  
Renal Transplant ◽  
Collagen Iv ◽  
Double Immunohistochemistry

scholarly journals In situ hybridization reveals temporal and spatial changes in cellular expression of mRNA for a laminin receptor, laminin, and basement membrane (type IV) collagen in the developing kidney.

1989 ◽  
Vol 109 (3) ◽  
pp. 1351-1362 ◽  
Author(s):  
G W Laurie ◽  
S Horikoshi ◽  
P D Killen ◽  
B Segui-Real ◽  
Y Yamada
Keyword(s):  
In Situ Hybridization ◽  
Steady State ◽  
Mrna Expression ◽  
Collagen Iv ◽  
Receptor Expression ◽  
Laminin Receptor ◽  
Northern Analysis ◽  
Cellular Expression ◽  
Distal Tubules

The appearance of extracellular matrix molecules and their receptors represent key events in the differentiation of cells of the kidney. Steady-state mRNA levels for a laminin receptor, the laminin B1, B2, and A chains, and the alpha 1-chain of collagen IV (alpha 1[IV]), were examined in mouse kidneys at 16 d gestation and birth, when cell differentiation is active, and 1-3 wk after birth when this activity has subsided. Northern analysis revealed that mRNA expression of laminin receptor precedes the alpha 1(IV) and laminin B chains whereas laminin A chain mRNA expression was very low. In situ hybridization reflected this pattern and revealed the cells responsible for expression. At 16 d gestation, laminin receptor mRNA was elevated in cells of newly forming glomeruli and proximal and distal tubules of the nephrogenic zone located in the kidney cortex. These cells also expressed mRNA for alpha 1(IV) and laminin chains. At birth, mRNA expression of receptor and all chains remained high in glomeruli but was reduced in proximal and distal tubules. At 1 wk after birth, expression was located in the medulla over collecting ducts and loops of Henle. Little expression was detectable by 3 wk. These results suggest that cellular expression of steady-state mRNA for laminin receptor, laminin, and collagen IV is temporally linked, with laminin receptor expression proceeding first and thereafter subsiding.

Origin of Urothelial Carcinoma After Renal Transplant Determined by Fluorescence In Situ Hybridization

2002 ◽  
Vol 167 (6) ◽  
pp. 2521-2522 ◽  
Author(s):  
VIRAJ A. MASTER ◽  
MAXWELL V. MENG ◽  
THERESA M. KOPPIE ◽  
PETER R. CARROLL ◽  
GARY D. GROSSFELD
Keyword(s):  
In Situ Hybridization ◽  
Renal Transplant ◽  
Fluorescence In Situ Hybridization ◽  
Urothelial Carcinoma

scholarly journals Accumulation of worn-out GBM material substantially contributes to mesangial matrix expansion in diabetic nephropathy

2017 ◽  
Vol 312 (6) ◽  
pp. F1101-F1111 ◽  
Author(s):  
Wilhelm Kriz ◽  
Jana Löwen ◽  
Giuseppina Federico ◽  
Jacob van den Born ◽  
Elisabeth Gröne ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Diabetic Nephropathy ◽  
In Situ Hybridization ◽  
Mesangial Cells ◽  
Collagen Iv ◽  
Mesangial Matrix ◽  
Electron Microscopic ◽  
Matrix Expansion ◽  
Production Mechanisms

Thickening of the glomerular basement membrane (GBM) and expansion of the mesangial matrix are hallmarks of diabetic nephropathy (DN), generally considered to emerge from different sites of overproduction: GBM components from podocytes and mesangial matrix from mesangial cells. Reevaluation of 918 biopsies with DN revealed strong evidence that these mechanisms are connected to each other, wherein excess GBM components fail to undergo degradation and are deposited in the mesangium. These data do not exclude that mesangial cells also synthesize components that contribute to the accumulation of matrix in the mesangium. Light, electron microscopic, immunofluorescence, and in situ hybridization studies clearly show that the thickening of the GBM is due not only to overproduction of components of the mature GBM (α3 and α5 chains of collagen IV and agrin) by podocytes but also to resumed increased synthesis of the α1 chain of collagen IV and of perlecan by endothelial cells usually seen during embryonic development. We hypothesize that these abnormal production mechanisms are caused by different processes: overproduction of mature GBM-components by the diabetic milieu and regression of endothelial cells to an embryonic production mode by decreased availability of mediators from podocytes.

Origin of Urothelial Carcinoma After Renal Transplant Determined by Fluorescence In Situ Hybridization

2002 ◽  
pp. 2521-2522
Author(s):  
VIRAJ A. MASTER ◽  
MAXWELL V. MENG ◽  
THERESA M. KOPPIE ◽  
PETER R. CARROLL ◽  
GARY D. GROSSFELD
Keyword(s):  
In Situ Hybridization ◽  
Renal Transplant ◽  
Fluorescence In Situ Hybridization ◽  
Urothelial Carcinoma

In Situ Hybridization for HPV in Drug-Induced Gingival Overgrowth in Renal Transplant Patients

2007 ◽  
Vol 103 (6) ◽  
pp. 790-791
Author(s):  
M. Magalhães ◽  
S. Alves-Júnior ◽  
M. Treiveiller ◽  
K. Ortega ◽  
N. Rezende
Keyword(s):  
In Situ Hybridization ◽  
Renal Transplant ◽  
Drug Induced ◽  
Gingival Overgrowth ◽  
Transplant Patients ◽  
Renal Transplant Patients

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

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