Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: KEGG:D02003 (NBT)
 1,323 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Tyrosine hydroxylase and phenoloxidase differ in that tyrosine hydroxylase (E.C.1.14.16.2) can hydroxylate tyrosine into -o-diphenol, but cannot oxidize the -o-diphenol, whereas phenoloxidase (E.C.1.14.18.1) is capable of oxidizing -o-diphenol to quinone. This difference can be exploited by staining tyrosine hydroxylase activity with a substrate-PMS-NBT method and staining the phenoloxidase with a dopamine-MBTH method. Based on the staining properties of the bands separated after electrophoresis, tyrosine hydroxylase has been differentiated from phenoloxidase in the silkworm Bombyx mori and the occurrence of tyrosine hydroxylase has been reported for the first time in this worm.
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PMID:Differentiation of tyrosine hydroxylase and phenol oxidase after electrophoresis. 750 28

The effect of a kappa-receptor agonist U-50, 488H on urine flow and the activity of TH-IR (tyrosine hydroxylase-immunoreactivity) neurons in PVH (Paraventricular neucleus of the hypothalamus) was studied in ethanol-anesthetized rats. U-50, 488H (5 micrograms/1 microliter) injected into PVH induced a potent diuretic response with no significant action on mean arterial pressure and heart rate. The diuretic response began within 20 min after injection, reached a peak value 41-60 min later and subsided at about 100 min. This diuretic response could be blocked by pretreatment with kappa-receptor antagonist NBT (Nor-Binaltorphimine Tetrahydrate). Both the number of neurons and staining density of TH-IR in the PVH tended to decrease at 20 min after the injection of U-50, 488H (10 micrograms/10 microliters) into the third cerebroventricle. The effect was most evident at 50 min and recovered at about 100 min after injection.
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PMID:[Effect of U-50, 488H administered centrally in rats on urination and activity of TH-IR neurons in PVH]. 765 95

We describe here a simple method for combining non-radioactive and radioactive in situ hybridization and immunohistochemistry on the same brain tissue section. This approach was first developed on the well-characterized hypothalamo-neurohypophyseal system, facilitating the optimization of the triple-labeling procedure and the verification of labeling specificity. We report the simultaneous detection of vasopressin (VP) mRNA with a digoxigenin-labeled oligonucleotide, oxytocin (OT) mRNA with a 35S-labeled oligonucleotide, and OT peptide in the same 12-microns cryostat section. This was performed on floating sections as follows: first, the two probes were hybridized simultaneously; second, the peptide was detected with an immunoperoxidase-DAB procedure; third, the digoxigenin-labeled probe was detected with an alkaline phosphatase-NBT/BCIP technique; and finally, the 35S-labeled probe was detected by histological autoradiography. We also demonstrate that this approach is suitable for the simultaneous detection of tyrosine hydroxylase and two less abundant mRNAs, vasoactive intestinal peptide and vasopressin mRNAs, in the suprachiasmatic nucleus. The combination of the three techniques did not significantly diminish their specificity or sensitivity. In conclusion, this new method, permitting the simultaneous detection of three different products of gene expression in the same section, could be useful for further analysis of the phenotypic organization and its plasticity in endocrine or neural tissues.
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PMID:Combination of non-radioactive and radioactive in situ hybridization with immunohistochemistry: a new method allowing the simultaneous detection of two mRNAs and one antigen in the same brain tissue section. 809 8

We have developed enhanced immunohistochemical protocols for detecting autonomic nerve fibers and splenocyte-associated proteins in rat spleen. This includes norepinephrine-synthesizing enzymes (dopamine-beta hydroxylase (DBH) and tyrosine hydroxylase (TH)), neuropeptide Y (NPY), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), c-fos protein, inducible nitric oxide synthase (iNOS), and the macrophage cell marker ED1. Animals were divided into sham-operated and splenic nerve-sectioned groups for detection of DBH, TH, and NPY. For immunodetection of TNF-alpha, iNOS, IFN-gamma and c-fos, animals were injected IV with saline or 100 microg of lipopolysaccharide (LPS) and were sacrificed at various time intervals post injection. Rats were perfused with 4% paraformaldehyde, spleens removed and cryoprotected, and 50-microm floating sections were cut on a freezing microtome. Immunodetection was performed with various detection systems and substrate/chromogen solutions, and in some cases using pretreatment with proteinase K (PK) for antigen unmasking. PK pretreatment increased immunostaining for DBH, TH, NPY, IFN-gamma, iNOS, and ED1, and the improvement was concentration-dependent. Using NPY immunostaining to index the signal-to-noise ratio for various substrates and detection systems, we found that an alkaline phosphatase detection system with NBT/BCIP as a substrate was the best procedure for light microscopy, whereas the CY3-labeled secondary antibody technique proved optimal for fluorescent microscopy. Surgical transection of the splenic nerve eliminated all nerve fiber staining for DBH, TH, and NPY. TNF-alpha, IFN-gamma, c-fos, and iNOS proteins were observed in the spleen in a time-dependent manner after LPS stimulation. Fluorescent double labeling, visualized with fluorescent confocal scanning laser microscopy, revealed many NPY fibers distributed among the ED1-labeled macrophages. These results demonstrate that immunohistochemistry can be used to index the activational effects of an immune challenge on splenocytes in situ and verifies that splenic immune cells are innervated by the sympathetic nervous system.
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PMID:Enhanced immunohistochemical detection of autonomic nerve fibers, cytokines and inducible nitric oxide synthase by light and fluorescent microscopy in rat spleen. 911 Dec 38