Renal Ischemia in Rats: Mitochondria Function and Laser Autofluorescence

Tirapelli, Luís Fernando; Bagnato, Vanderlei Salvador; Tirapelli, Daniela P.C.; Kurachi, Cristina; Barione, D.F.; Tucci, Sílvio; Suaid, Haylton Jorge; Cologna, Adauto José; Martins, Antônio Carlos Pereira

doi:10.1016/j.transproceed.2008.02.081

Cited by 12 publications

(10 citation statements)

References 35 publications

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“…[1][2][3][4] Kidney tissue induces high levels of AF, and the intensity of AF has been used to assess severity of tubular necrosis 5 and the degree of damage in ischemic kidney. [6][7][8] However, AF is an obstacle to immunofluorescence (IF) analysis that can either mask or interfere with specific fluorescent signals, 9,10 especially those with weak IF labeling, including direct IF and multiple-color fluorescence staining. In addition to intrinsic fluorescence of kidney is AF that arises from the tissue-processing techniques, including fixation agents such as glutaraldehyde 11 and embedding material such as paraffin.…”

mentioning

confidence: 99%

Sudan Black B Reduces Autofluorescence in Murine Renal Tissue

Sun¹,

Hong²,

Zheng³

et al. 2011

Archives of Pathology &Amp; Laboratory Medicine

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N Context.-Renal tissue emits intense autofluorescence, making it difficult to differentiate specific immunofluorescence signals and thus limiting its application to clinical biopsy material.Objective.-To identify and minimize autofluorescence of renal tissue and demonstrate a simple, efficient method to reduce autofluorescence using Sudan black B.Design.-In this study, the sources and features of autofluorescence emitted from kidney tissue were examined. Broad autofluorescence was visualized in both frozen and paraffin kidney sections of normal mice and mice with Adriamycin-induced nephropathy using confocal laser scanning microscopy. Autofluorescence appeared in commonly used 49,6-diamidino-2-phenylindole, fluorescein isothiocyanate, and Texas Red channels but not in far-red channel, and emitted extensively from red cells, injured tubulointersitial cells, and protein casts in diseased kidney.To eliminate autofluorescence, Sudan black B was used on formaldehyde-fixed paraffin sections and frozen sections of mouse kidney. The effects of Sudan black B in various concentrations were tested on kidney tissue.Results.-The 0.1% Sudan black B effectively blocked autofluorescence from both paraffin and frozen sections without adversely affecting specific fluorescence signals. Interestingly, the solvent for Sudan black B, 70% ethanol, was also shown to reduce autofluorescence on frozen sections, but not on paraffin sections.Conclusions.-This study demonstrates a simple, efficient, and cost-effective method to reduce autofluorescence using Sudan black B, and also provides a comprehensive approach to identify and minimize autofluorescence of renal tissue.

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confidence: 99%

Sudan Black B Reduces Autofluorescence in Murine Renal Tissue

Sun¹,

Hong²,

Zheng³

et al. 2011

Archives of Pathology &Amp; Laboratory Medicine

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“…Blood was collected from the inferior vena cava of anaesthetized rats (thiopental sodium, 0.4 mg/kg, i.p.) using heparinzed syringes, and the samples were analysed using a CG‐17A gas chromatographer (Shimadzu, Kyoto, Japan) as previously described [12,22] . The results are expressed as mg ethanol/ml blood.…”

Section: Methodsmentioning

confidence: 99%

“…Kidney mitochondria were prepared in 0.25 mol/l sucrose and 1 mmol/l ethylenediamine tetraacetic acid at pH 7.2 and 4°C using standard centrifugation procedures. Mitochondrial protein and mitochondrial respiratory function were measured as previously described [22] . Mitochondrial respiration was initiated by the addition of succinate (5 mmol/l final concentration) and oxidative phosphorylation by the addition of 200 mmol/l ADP.…”

Section: Methodsmentioning

confidence: 99%

“…For histology, tissue samples fixed in 10% buffered formalin were paraffin embedded for preparation of 5‐µm sections that were stained with hematoxylin and eosin as previously described [22] . Using a binocular Zeiss microscope (model Axioskop 2 plus, Jena, Germany), the interstitial tubular damage was graded according to Goujon criteria, [23] which analyses six basic morphological patterns.…”

Section: Methodsmentioning

confidence: 99%

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Ethanol consumption increases the expression of endothelial nitric oxide synthase, inducible nitric oxide synthase and metalloproteinases in the rat kidney

Tirapelli

Martins-Oliveira

Batalhão³

et al. 2012

Journal of Pharmacy and Pharmacology

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“…Exposure to hypoxia leads to accumulation of highly reduced metabolic intermediates in mitochondria, which, upon reoxygenation, can result in a surge of ROS production and damage mitochondrial proteins, lipids and DNA (Piper et al, 2003;Sadek et al, 2003;Zenebe et al, 2007). The ROS-induced mitochondrial damage is critically involved in ischemia-reperfusion injury during stroke, infarction, and tissue and organ transplantation (Levraut et al, 2003;Piper et al, 2003;Sadek et al, 2002Sadek et al, , 2003Tirapelli et al, 2008). However, some animals such as diving birds and mammals or intertidal invertebrates can undergo frequent H/R cycles without any apparent tissue damage.…”

Section: Introductionmentioning

confidence: 99%

Effects of intermittent hypoxia on oxidative stress and protein degradation in molluscan mitochondria

Ivanina

Sokolova

2016

Journal of Experimental Biology

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Oxygen fluctuations represent a common stressor in estuarine and intertidal environments and can compromise the mitochondrial integrity and function in marine organisms. We assessed the role of mitochondrial protection mechanisms (ATP-dependent and -independent mitochondrial proteases, and antioxidants) in tolerance to intermittent hypoxia or anoxia in three species of marine bivalves: hypoxia-tolerant hard clams (Mercenaria mercenaria) and oysters (Crassostrea virginica), and a hypoxia-sensitive subtidal scallop (Argopecten irradians). In clams and oysters, mitochondrial tolerance to hypoxia (18 h at 5% O 2 ), anoxia (18 h at 0.1% O 2 ) and subsequent reoxygenation was associated with the ability to maintain the steadystate activity of ATP-dependent and -independent mitochondrial proteases and an anticipatory upregulation of the total antioxidant capacity under the low oxygen conditions. No accumulation of endproducts of lipid or protein peroxidation was found during intermittent hypoxia or anoxia in clams and oysters (except for an increase in protein carbonyl concentration after hypoxia-reoxygenation in oysters). In contrast, hypoxia/anoxia and reoxygenation strongly suppressed activity of the ATP-dependent mitochondrial proteases in hypoxiasensitive scallops. This suppression was associated with accumulation of oxidatively damaged mitochondrial proteins (including carbonylated proteins and proteins conjugated with a lipid peroxidation product malondialdehyde) despite high total antioxidant capacity levels in scallop mitochondria. These findings highlight a key role of mitochondrial proteases in protection against hypoxia-reoxygenation stress and adaptations to frequent oxygen fluctuations in intertidal mollusks.

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Renal Ischemia in Rats: Mitochondria Function and Laser Autofluorescence

Cited by 12 publications

References 35 publications

Sudan Black B Reduces Autofluorescence in Murine Renal Tissue

Sudan Black B Reduces Autofluorescence in Murine Renal Tissue

Ethanol consumption increases the expression of endothelial nitric oxide synthase, inducible nitric oxide synthase and metalloproteinases in the rat kidney

Effects of intermittent hypoxia on oxidative stress and protein degradation in molluscan mitochondria

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