Detecting necrotic neurons with fluoro-jade stain

Krinke, G.; Classen, W.; Vidotto, N; Suter, Eugénie E.; Würmlin, C H

doi:10.1078/0940-2993-00202

Cited by 37 publications

(20 citation statements)

References 14 publications

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“…Fluoro-Jade fluorochromes label neurons at a late stage in degeneration, at a time when cell loss occurs, and are a marker of neuronal death irrespective of the type of cell death (apoptosis or necrosis) involved (Krinke et al, 2001;Rocha et al, 2004;Schumed and Hopkins, 2000;Seoane et al, 2005). The labeling of the positive control samples from rats treated with trans-crotononitrile (Boadas-Vaello et al, 2005) demonstrated that the staining process was performed adequately.…”

Section: Discussionmentioning

confidence: 99%

Dose-dependent regional brain acetylcholinesterase and acylpeptide hydrolase inhibition without cell death after chlorpyrifos administration

et al. 2013

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-Organophosphates (OPs) are important toxic compounds commonly used for a variety of purposes in agriculture, industry and household settings. It has been well established that the main mechanism of acute toxic action of OP is the inhibition of acetylcholinesterase (AChE). However, we observed long term deficit after acute subcutaneous exposure to Chlorpyrifos (CPF) even when AChE activity is restored. In fact, besides AChE inhibition, non-AChE targets have also been proposed as an alternative mechanism involved in the acute lethal action and side effects of short or long-term exposure. In this context, our main aim in this research was to establish a dose-response curve of Acylpeptide hydrolase (APH) and AChE regional brain activity after acute CPF administration that could explain these long term effects observed in the literature. Moreover, since available data suggest that long term effects of OPs exposure could involve neuronal cell death, our second aim was to evaluate, assessing by Fluoro-Jade B (FJB) staining, whether CPF produces induced cell death. Our results show that an acute exposure to 250 mg/kg CPF does not induce neuronal death as measured by FJB but produces highest AChE regional brain inhibition after administration. In addition, APH seems to be more sensitive than AChE to CPF exposure because after 31 days of exposure, complete recovery was seen only for APH activity at Frontal Cortex, Cerebellum and Brain Stem.

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Section: Discussionmentioning

confidence: 99%

Dose-dependent regional brain acetylcholinesterase and acylpeptide hydrolase inhibition without cell death after chlorpyrifos administration

et al. 2013

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“…Examples of likely ancillary methods include conventional neurohistology stains, such as cresyl violet (to reveal fine neuronal cytoarchitecture) and Luxol fast blue (to view myelin integrity); various immunohistochemical biomarkers of cell origin, like glial fibrillary acidic protein (to detect reactive astrocytes at sites of subacute to chronic brain injury) and ionized calcium binding adaptor molecule 1 (Iba1, to reveal microglia); and techniques to detect neuronal degeneration, such as amino cupric silver, Fluoro-Jade B, or Fluoro-Jade C. The recent literature contains many superb articles that address the rationale and technical considerations for deploying these special procedures (Fix and Garman 2000;Fix et al 1996;Barone et al 2000;Krinke et al 2001;Switzer and Butt 2011;Schmued et al 2005). …”

Section: Neurohistology Methodsmentioning

confidence: 99%

STP Position Paper

Garman²,

et al. 2013

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The Society of Toxicologic Pathology charged a Nervous System Sampling Working Group with devising recommended practices to routinely screen the central nervous system (CNS) and peripheral nervous system (PNS) in Good Laboratory Practice-type nonclinical general toxicity studies. Brains should be weighed and trimmed similarly for all animals in a study. Certain structures should be sampled regularly: caudate/putamen, cerebellum, cerebral cortex, choroid plexus, eye (with optic nerve), hippocampus, hypothalamus, medulla oblongata, midbrain, nerve, olfactory bulb (rodents only), pons, spinal cord, and thalamus. Brain regions may be sampled bilaterally in rodents using 6 to 7 coronal sections, and unilaterally in nonrodents with 6 to 7 coronal hemisections. Spinal cord and nerves should be examined in transverse and longitudinal (or oblique) orientations. Most Working Group members considered immersion fixation in formalin (for CNS or PNS) or a solution containing acetic acid (for eye), paraffin embedding, and initial evaluation limited to hematoxylin and eosin (H&E)-stained sections to be acceptable for routine microscopic evaluation during general toxicity studies; other neurohistological methods may be undertaken if needed to better characterize H&E findings. Initial microscopic analyses should be qualitative and done with foreknowledge of treatments and doses (i.e., ''unblinded''). The pathology report should clearly communicate structures that were assessed and methodological details. Since neuropathologic assessment is only one aspect of general toxicity studies, institutions should retain flexibility in customizing their sampling, processing, analytical, and reporting procedures as long as major neural targets are evaluated systematically.

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“…In general, peripheral nerves in DNT studies are routinely taken from the hind limb and include the sciatic, tibial (internal popliteal), common peroneal (external popliteal, fibular), sural, and plantar nerves (Figure 4). In our experience, all these levels must be sampled due to inherent differences in the susceptibility of various regions of nerve from proximal to distal to toxicant-induced damage (Krinke et al, 1979(Krinke et al, , 2001. Some pathologists prefer to post-fix the nerves while they remain in the carcass, while others remove nerves immediately after perfusion fixation and staple them (through their ends) to a flat card to facilitate post-fixation.…”

Section: Tissue Trimming and Sectioningmentioning

confidence: 99%

“…Common sites for sampling skeletal muscle are the gastrocnemius muscles (a recommended site based on its known anatomic and physiological characteristics, such as the presence of intramuscular tibial nerve branches that are known to be highly sensitive to neurotoxicity; Krinke et al, 1979Krinke et al, , 2001, the diaphragm, and the biceps femoris muscle. Evaluation of fixed muscle tissue allows agent-induced structural changes to be defined in both muscular and neural elements.…”

Section: Fixation and Tissue Collectionmentioning

confidence: 99%

A ‘Best Practices’ Approach to Neuropathologic Assessment in Developmental Neurotoxicity Testing—for Today

Bolon

Garman²,

Jensen

et al. 2006

Toxicol Pathol

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A key trait of developmental neurotoxicants is their ability to cause structural lesions in the immature nervous system. Thus, neuropathologic assessment is an essential element of developmental neurotoxicity (DNT) studies that are designed to evaluate chemically-induced risk to neural substrates in young humans. The guidelines for conventional DNT assays have been established by regulatory agencies to provide a flexible scaffold for conducting such studies; recent experience has launched new efforts to update these recommendations. The present document was produced by an ad hoc subcommittee of the Society of Toxicologic Pathology (STP) tasked with examining conventional methods used in DNT neuropathology in order to define the 'best practices' for dealing with the diverse requirements of both national (EPA) and international (OECD) regulatory bodies. Recommendations (including citations for relevant neurobiological and technical references) address all aspects of the DNT neuropathology examination: study design; tissue fixation, collection, processing, and staining; qualitative and quantitative evaluation; statistical analysis; proper control materials; study documentation; and personnel training. If followed, these proposals will allow pathologists to meet the need for a sound risk assessment (balanced to address both regulatory issues and scientific considerations) in this field today while providing direction for the research needed to further refine DNT neuropathology 'best practices' in the future.

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Detecting necrotic neurons with fluoro-jade stain

Cited by 37 publications

References 14 publications

Dose-dependent regional brain acetylcholinesterase and acylpeptide hydrolase inhibition without cell death after chlorpyrifos administration

Dose-dependent regional brain acetylcholinesterase and acylpeptide hydrolase inhibition without cell death after chlorpyrifos administration

STP Position Paper

A ‘Best Practices’ Approach to Neuropathologic Assessment in Developmental Neurotoxicity Testing—for Today

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