Fluoro Nissl Green: a novel fluorescent counterstain for neuroanatomy

Quinn, Bruce; Toga, Arthur W.; Motamed, Soheil; Merlic, Craig A.

doi:10.1016/0304-3940(94)11198-r

“…Neuronal staining was carried out through Nissl staining with NeuroTrace 500/525 green (Invitrogen, no. N21480), which binds to negatively charged nucleic acids in the ER (endoplasmic reticulum) of neurons (Quinn et al, 1995). We used a monoclonal RIP (receptor-interacting protein) antibody (Millipore, no.…”

Section: Methodsmentioning

confidence: 99%

Nanoparticle Targeting to Neurons in a Rat Hippocampal Slice Culture Model

Walters

¹

,

Kraig

²

,

Medintz

³

et al. 2012

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We have previously shown that CdSe/ZnS core/shell luminescent semiconductor nanocrystals or QDs (quantum dots) coated with PEG [poly(ethylene glycol)]-appended DHLA (dihydrolipoic acid) can bind AcWG(Pal)VKIKKP9GGH6 (Palm1) through the histidine residues. The coating on the QD provides colloidal stability and this peptide complex uniquely allows the QDs to be taken up by cultured cells and readily exit the endosome into the soma. We now show that use of a polyampholyte coating [in which the neutral PEG is replaced by the negatively heterocharged CL4 (compact ligand)], results in the specific targeting of the palmitoylated peptide to neurons in mature rat hippocampal slice cultures. There was no noticeable uptake by astrocytes, oligodendrocytes or microglia (identified by immunocytochemistry), demonstrating neuronal specificity to the overall negatively charged CL4 coating. In addition, EM (electron microscopy) images confirm the endosomal egress ability of the Palm1 peptide by showing a much more disperse cytosolic distribution of the CL4 QDs conjugated to Palm1 compared with CL4 QDs alone. This suggests a novel and robust way of delivering neurotherapeutics to neurons.

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“…Fluorescent Nissl staining was performed as described previously (Quinn et al, 1995) using the NeuroTrace 500/525 green fluorescent Nissl stain (Invitrogen). Briefly, rehydrated slices were incubated for 20 min with 1:100 diluted NeuroTrace.…”

Section: Methodsmentioning

confidence: 99%

Cholinergic Control in Developing Prefrontal–Hippocampal Networks

Janiesch¹,

Krüger²,

Poschel³

et al. 2011

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The cholinergic drive enhances input processing in attentional and mnemonic context by interacting with the activity of prefrontalhippocampal networks. During development, acetylcholine modulates neuronal proliferation, differentiation, and synaptic plasticity, yet its contribution to the maturation of cognitive processing resulting from early entrainment of neuronal networks in oscillatory rhythms remains widely unknown. Here we show that cholinergic projections growing into the rat prefrontal cortex (PFC) toward the end of the first postnatal week boost the generation of nested gamma oscillations superimposed on discontinuous spindle bursts by acting on functional muscarinic but not nicotinic receptors. Although electrical stimulation of cholinergic nuclei increased the occurrence of nested gamma spindle bursts by 41%, diminishment of the cholinergic input by either blockade of the receptors or chronic immunotoxic lesion had the opposite effect. This activation of locally generated gamma episodes by direct cholinergic projections to the PFC was accompanied by indirect modulation of underlying spindle bursts via cholinergic control of hippocampal theta activity. With ongoing maturation and switch of network activity from discontinuous bursts to continuous theta-gamma rhythms, accumulating cholinergic projections acting on both muscarinic and nicotinic receptors mediated the transition from high-amplitude slow to low-amplitude fast rhythms in the PFC. By exerting multiple actions on the oscillatory entrainment of developing prefrontal-hippocampal networks, the cholinergic input may refine them for later gating processing in executive and mnemonic tasks.

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“…The samples were incubated with secondary antibody followed by DAB treatment. Slides were counter-stained with fluorescent Nissl reagent to enable identification of intact neurons by presence of the Nissl substance [27]. …”

Section: Methodsmentioning

confidence: 99%

Cytosolic phospholipase A2alpha amplifies early cyclooxygenase-2 expression, oxidative stress and MAP kinase phosphorylation after cerebral ischemia in mice

Kishimoto

¹

,

Li

²

,

Zhang

³

et al. 2010

J Neuroinflammation

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Background The enzyme cytosolic phospholipase A 2 alpha (cPLA 2 α) has been implicated in the progression of cerebral injury following ischemia and reperfusion. Previous studies in rodents suggest that cPLA 2 α enhances delayed injury extension and disruption of the blood brain barrier many hours after reperfusion. In this study we investigated the role of cPLA 2 α in early ischemic cerebral injury. Methods Middle cerebral artery occlusion (MCAO) was performed on cPLA 2 α +/+ and cPLA 2 α -/- mice for 2 hours followed by 0, 2, or 6 hours of reperfusion. The levels of cPLA 2 α, cyclooxygenase-2, neuronal morphology and reactive oxygen species in the ischemic and contralateral hemispheres were evaluated by light and fluorescent microscopy. PGE 2 content was compared between genotypes and hemispheres after MCAO and MCAO and 6 hours reperfusion. Regional cerebral blood flow was measured during MCAO and phosphorylation of relevant MAPKs in brain protein homogenates was measured by Western analysis after 6 hours of reperfusion. Results Neuronal cPLA 2 α protein increased by 2-fold immediately after MCAO and returned to pre-MCAO levels after 2 hours reperfusion. Neuronal cyclooxygenase-2 induction and PGE 2 concentration were greater in cPLA 2 α +/+ compared to cPLA 2 α -/- ischemic cortex. Neuronal swelling in ischemic regions was significantly greater in the cPLA 2 α +/+ than in cPLA 2 α -/- brains (+/+: 2.2 ± 0.3 fold vs. -/-: 1.7 ± 0.4 fold increase; P < 0.01). The increase in reactive oxygen species following 2 hours of ischemia was also significantly greater in the cPLA 2 α +/+ ischemic core than in cPLA 2 α -/- (+/+: 7.12 ± 1.2 fold vs. -/-: 3.1 ± 1.4 fold; P < 0.01). After 6 hours of reperfusion ischemic cortex of cPLA 2 α +/+ , but not cPLA 2 α -/- , had disruption of neuron morphology and decreased PGE 2 content. Phosphorylation of the MAPKs-p38, ERK 1/2, and MEK 1/2-was significantly greater in cPLA 2 a +/+ than in cPLA 2 α -/- ischemic cortex 6 hours after reperfusion. Conclusions ...

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Fluoro Nissl Green: a novel fluorescent counterstain for neuroanatomy

Cited by 26 publications

References 10 publications

Nanoparticle Targeting to Neurons in a Rat Hippocampal Slice Culture Model

Nanoparticle Targeting to Neurons in a Rat Hippocampal Slice Culture Model

Cholinergic Control in Developing Prefrontal–Hippocampal Networks

Cytosolic phospholipase A2alpha amplifies early cyclooxygenase-2 expression, oxidative stress and MAP kinase phosphorylation after cerebral ischemia in mice

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