A Microwave Applicator for in Vivo Rapid Inactivation of Enzymes in the Central Nervous System (Short Papers)

Lenox, Robert H.; Gandhi, O.P.; Meyerhoff, James L.; Grove, H.M.

doi:10.1109/tmtt.1976.1128770

Cited by 42 publications

(10 citation statements)

References 12 publications

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“…Sacrificing rats or mice by focused microwave irradiation results in rapid heat inactivation of brain enzymes (Lenox et al, 1976;Stavinoha, 1993). This technique has been used to preserve in vivo levels of a variety of neurotransmitters (Butcher et al, 1976;Ishikawa et al, 1982;Katsura et al, Abbreviations: ATF, activating transcription factor; BCA, bicinchoninic acid; CAMK, calcium/calmodulin-dependent kinase; CREB, cyclic AMP response element-binding protein; ERK, extracellular signal-regulated kinase; GFAP, glial fibrillary acidic protein; HRP, horse radish peroxidase; MAPK, mitogen activated protein kinase; PAGE, polyacrylamide gel electrophoresis; PBS, phosphate buffered saline; SAPK, stress-activated protein kinase; SDS, sodium dodecyl sulfate; STAT, signal transducer and activator of transcription * Corresponding author.…”

Section: Introductionmentioning

confidence: 99%

Focused microwave irradiation of the brain preserves in vivo protein phosphorylation: comparison with other methods of sacrifice and analysis of multiple phosphoproteins

O’Callaghan

Sriram

2004

Journal of Neuroscience Methods

100

110

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Section: Introductionmentioning

confidence: 99%

Focused microwave irradiation of the brain preserves in vivo protein phosphorylation: comparison with other methods of sacrifice and analysis of multiple phosphoproteins

O’Callaghan

Sriram

2004

Journal of Neuroscience Methods

100

110

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“…; (2) theoretical and practical studies on the microwave fixation method, including basic studies of protein cross-linking mechanism and the heating effect of MWI on biological materials (3, 6-10,17, 20-27, 36-39, 46, 49-51, 55, 63, 63, 70, 72, 82, 84-86, 93-99); (3) studies on the bio-and histochemical stabilization of enzymes in the central nervous tissues by MWI, especially extremely strong irradiation of more than 10 KW is used to fix brain tissue (11,27,31,44,55,65,67,68,(89)(90)(91)(92)(93)96).…”

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confidence: 99%

New tissue fixation method for cytochemistry using migrowave irradiation. I. General remarks.

Mizuhira

Notoya

Hasegawa

1990

Acta Histochem. Cytochem

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We examined a rapid tissue fixation method using microwave irradiation (MWI) for ultrastructural, and cytochemical studies, including those on enzymeor immunolectin-cytochemistry, autoradiography, and X-ray microanalysis. MWI was applied to 0.3 to 1 cm3 blocks of several kinds of rat tissues at 2.45 GHz/500 W for about 20 sec in a fixative, as shown in the text, at room temperature.We tried to find why MWI enabled the aldehyde to penetrate into the tissue block so quickly. Microautoradiography was performed using 3H-formalin as a tracer. With MWI, prominently developed silver grains appeared homogeneously in the section, while the control section showed no silver grains within the section, only around it. Clearly, the microwave energy enables 3H-formalin to penetrate quickly into the tissue block. However, using biochemical analysis, we found that complete tissue fixation occurred only after the chemical crosslinking of proteins with aldehyde which was a gradual process. Thus, for good fixation by MWI, the tissue blocks needed to be left in the fixation medium for about 10 to 60 min after MWI at room temperature for cytochemical fixation and for about 30 to 180 min for conventional fixation.A fixative containing a small amount of tannic acid, 0.05 to 0.1%, is effective for fixing soluble peptides or proteins. A fixative containing NHA (see in the text), phosphate or oxalate can be used to detect the localization of Ca ion in tissue and cells. Combinations of these techniques have yielded excellent results in conventional light and electron microscopy, as well as in histochemical and cytochemical studies.MWI also yields very good results in high resolution electron microscopy, for example, many cholinergic synaptic vesicles at the presynaptic membrane in the rat brain could be observed for their opening images. The fixation effect of MWI seems to be equivalent to that of the rapid freezing fixation method.The enzymatic activity of alkaline phosphatase could withstand MWI treatment, even when the sample had been embedded in Technovit resin. However, acid phosphatase and glucose-6-phosphatase seemed to be rather weak against the MW!. Lectin binding sites also were stable against MWI treatment. MWI is excellent for micro-and electron autoradiography and also for X-ray microanalysis.

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“…The uncharacterized peptide was found to be processed at dibasic amino acid sites from the known neuropeptide containing secretogranin 1 precursor. temperature to 90 • C, the delay until enzyme inactivation is avoided [50,55,56]. The proteolytic events are thereby inactivated by irreversible heat denaturation and, in contrast with freezing methods, they permit further dissection and processing of the sample [57].…”

Section: Sample Preparationmentioning

confidence: 99%

Neuropeptidomics: expanding proteomics downwards

Svensson

Sköld

Nilsson

et al. 2007

Biochemical Society Transactions

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Biological function is mainly carried out by a dynamic population of proteins and peptides which may be used as markers for disease diagnosis, prognosis and as a guide for effective treatment. The study of proteins is called proteomics and it is generally performed by two-dimensional gel electrophoresis and mass spectrometric methods. However, gel-based proteomics is methodologically restricted from the low mass region, which includes important endogenous peptides. The study of endogenous peptides, peptidomics, is complicated by protein fragments produced post-mortem during conventional sample handling. Nanoflow liquid chromatography and MS, together with improved methods for sample preparation, have been used to semi-quantitatively monitor endogenous peptides in brain tissue. When rapidly heat-denatured brain tissue was analysed, these methods enabled simultaneous detection of hundreds of peptides and the identification of several endogenous peptides not previously described in the literature. In an application of the MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) model for Parkinson's disease, the expression of the small protein PEP-19 was compared with controls. The levels were found to be significantly decreased in the striatum of MPTP-treated animals.

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A Microwave Applicator for in Vivo Rapid Inactivation of Enzymes in the Central Nervous System (Short Papers)

Cited by 42 publications

References 12 publications

Focused microwave irradiation of the brain preserves in vivo protein phosphorylation: comparison with other methods of sacrifice and analysis of multiple phosphoproteins

Focused microwave irradiation of the brain preserves in vivo protein phosphorylation: comparison with other methods of sacrifice and analysis of multiple phosphoproteins

New tissue fixation method for cytochemistry using migrowave irradiation. I. General remarks.

Neuropeptidomics: expanding proteomics downwards

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