Traumatic Brain Injury-Induced Ependymal Ciliary Loss Decreases Cerebral Spinal Fluid Flow

Xiong, Guoxiang; Elkind, Jaclynn A.; Kundu, Suhali; Smith, Colin J.; Antunes, Marcelo B.; Tamashiro, Edwin; Kofonow, Jennifer M.; Mitala, Christina M.; Cole, John T.; Stein, Sherman C.; Grady, M. Sean; Einhorn, Eugene; Cohen, Noam A.; Cohen, Akiva S.

doi:10.1089/neu.2013.3110

Cited by 33 publications

(48 citation statements)

References 59 publications

(83 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In a previous study 16 and the present report, we showed that methanol treatment is necessary for immunofluorescent and immunogold staining of ependymal basal bodies in paraformaldehyde-fixed brain slices. With the present optimization, treatment with methanol at the concentration from 33.3% to 75% can result in ideal staining of basal bodies without inducing obvious deformations to the brain slices.…”

Section: Discussionsupporting

confidence: 62%

“…The rabbit polyclonal antibody against γ-tubulin was used to stain out the basal bodies of ependymal cells, as shown previously. 16 Before immunostaining, we applied different concentration of methanol to regular vibratome slices at RT for 30 minutes or to sucrose-protected slices at −20°C for 10 minutes. From the concentration of 33.3% to 100%, methanol treatment at both temperature conditions resulted in a similar immunofluorescent staining pattern of basal bodies in these brain slices (Figs.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Optimized Protocol of Methanol Treatment for Immunofluorescent Staining in Fixed Brain Slices

Yuan¹,

Xiong

Cohen³

et al. 2017

Applied Immunohistochemistry &Amp; Molecular Morphology

Self Cite

View full text Add to dashboard Cite

We optimized methanol treatment in paraformaldehyde-fixed slices for immunofluorescent staining of ependymal basal bodies in brain ventricles. As 100% methanol induced severe deformations to the slices (including rolling and folding over), we tried to decrease methanol concentration. We found that 33.3% to 75% methanol could result in ideal immunostaining of basal bodies without inducing obvious deformations. Instead of treating slices at À 201C (without proper cryoprotection measurements) as suggested in previous studies, we carried out methanol treatment at room temperature. Our modified protocol can not only raise immunostaining efficiency in tissue slices, it may also prevent potential freezing damages to the samples. I mmunohistochemical staining efficiency largely depends on tissue processing. 1-4 As a tissue processing reagent, methanol has been widely used to fix or permeabilize cultured cells. [5][6][7][8] It was also used to treat tissue blocks and slices. 9,10 To achieve the best immunostaining, cells need to be treated with 100% methanol at À 201C for 10 to 20 minutes. [10][11][12] We noticed that none of these studies or protocols adopted proper cryoprotection measurements. Without effective cryoprotection, treatment at À 201C might bring freezing damages to biochemical identities and/or morphologic features of the cells. 3 These morphologic and biochemical alterations may result in inconsistent immunostaining results, 13 as suggested in another study. 14 In the present study, we checked the effects of methanol treatment on paraformaldehyde-fixed brain slices before immunofluorescent staining. To our surprise, 100% methanol would result in severe deformations to the slices including rolling and folding over. We then tested various concentrations of methanol to the slices in different conditions. MATERIALS AND METHODSA total of 12 C57/Bl6 male mice (Jackson Laboratory, Bar Harbor, ME) at 6 to 8 weeks were used. The procedures and protocols for all animal studies were approved by Institutional Animal Care and Use Committees of Tongji University, Children's Hospital of Philadelphia, and University of Pennsylvania, in accordance with international guidelines on the ethical use of animals (National Research Council, 1996). After the mice were deeply anesthetized and perfused, the brains were removed and postfixed, and then vibratome slices were cut at 50 mm and collected in 6 serials in PBS as reported previously. 15 The slices were treated with different concentrations of methanol (0%, 33.3%, 50%, 75%, 95%, and 100%) at room temperature (RT) for 30 minutes or at À 201C for 10 minutes. Before methanol treatment at À 201C, slices were immersed in 20% sucrose overnight and then 30% sucrose for 2 days at 41C, except for control ones. The slices were then permabilized with 0.3% triton X-100 and blocked with a mixture of 5% normal goat serum and 1% bovine serum albumin before free-floating immunostaining.For double immunofluorescent staining, we incubated the slices with a mouse monoclonal antibody ...

show abstract

Section: Discussionsupporting

confidence: 62%

Section: Resultsmentioning

confidence: 99%

Optimized Protocol of Methanol Treatment for Immunofluorescent Staining in Fixed Brain Slices

Yuan¹,

Xiong

Cohen³

et al. 2017

Applied Immunohistochemistry &Amp; Molecular Morphology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Intravital particle tracking has been used to indirectly observe blood flow velocity with tracer particles in the systemic circulation [107-110] and in the colonic microcirculation in experimental inflammatory bowel disease (IBD) [111], as well as to indirectly observe decreases in cerebrospinal fluid flow after traumatic brain injury [112]. More recently, intravital particle tracking techniques have been adapted to tracking NP for drug and gene delivery in vivo in real time.…”

Section: The Utility Of Particle Tracking Analysis In Drug and Genmentioning

confidence: 99%

Particle tracking in drug and gene delivery research: State-of-the-art applications and methods

Schuster

Ensign

Allan

et al. 2015

Advanced Drug Delivery Reviews

122

139

View full text Add to dashboard Cite

Particle tracking is a powerful microscopy technique to quantify the motion of individual particles at high spatial and temporal resolution in complex fluids and biological specimens. Particle tracking's applications and impact in drug and gene delivery research have greatly increased during the last decade. Thanks to advances in hardware and software, this technique is now more accessible than ever, and can be reliably automated to enable rapid processing of large data sets, thereby further enhancing the role that particle tracking will play in drug and gene delivery studies in the future. We begin this review by discussing particle tracking-based advances in characterizing extracellular and cellular barriers to therapeutic nanoparticles and in characterizing nanoparticle size and stability. To facilitate wider adoption of the technique, we then present a user-friendly review of state-of-the-art automated particle tracking algorithms and methods of analysis. We conclude by reviewing technological developments for next-generation particle tracking methods, and we survey future research directions in drug and gene delivery where particle tracking may be useful.

show abstract

“…Ependymal cells are specialized glia, that line the ventricles of the CNS. Ependymal cell lose was found to decrease ventricular flow following TBI which could negatively affect the waste and nutrient exchange within the brain (Xiong et al, 2014 ). Additional research that helps decipher the molecular pathways between glia and memory deficits will be vital for development of better clinical tools for gauging memory loss.…”

Section: Glial Contribution To Memory Deficitsmentioning

confidence: 99%

Role of Glia in Memory Deficits Following Traumatic Brain Injury: Biomarkers of Glia Dysfunction

Sajja

Hlavac

VandeVord

2016

Front. Integr. Neurosci.

View full text Add to dashboard Cite

Historically, glial cells have been recognized as a structural component of the brain. However, it has become clear that glial cells are intimately involved in the complexities of neural networks and memory formations. Astrocytes, microglia, and oligodendrocytes have dynamic responsibilities which substantially impact neuronal function and activities. Moreover, the importance of glia following brain injury has come to the forefront in discussions to improve axonal regeneration and functional recovery. The numerous activities of glia following injury can either promote recovery or underlie the pathobiology of memory deficits. This review outlines the pathological states of glial cells which evolve from their positive supporting roles to those which disrupt synaptic function and neuroplasticity following injury. Evidence suggests that glial cells interact extensively with neurons both chemically and physically, reinforcing their role as pivotal for higher brain functions such as learning and memory. Collectively, this mini review surveys investigations of how glial dysfunction following brain injury can alter mechanisms of synaptic plasticity and how this may be related to an increased risk for persistent memory deficits. We also include recent findings, that demonstrate new molecular avenues for clinical biomarker discovery.

show abstract

Traumatic Brain Injury-Induced Ependymal Ciliary Loss Decreases Cerebral Spinal Fluid Flow

Cited by 33 publications

References 59 publications

Optimized Protocol of Methanol Treatment for Immunofluorescent Staining in Fixed Brain Slices

Optimized Protocol of Methanol Treatment for Immunofluorescent Staining in Fixed Brain Slices

Particle tracking in drug and gene delivery research: State-of-the-art applications and methods

Role of Glia in Memory Deficits Following Traumatic Brain Injury: Biomarkers of Glia Dysfunction

Contact Info

Product

Resources

About