Neurodegeneration - Science topic

Dr. He, the metabolism of the biomarkers of neurodegeneration and the impact of hepatic and renal dysfunctions seem to have been little explored. What about

Suhocki PV, Doraiswamy PM. Cerebral venous biomarkers and veno-arterial gradients: untapped resources in Alzheimer’s disease. Front Neurol 04 January 2023; 14.  https://doi.org/10.3389/fneur.2023.1295122 ...

or exploring diagnostic links to

Herndon RM, Johnson M. A method for the electron microscopic study of cerebrospinal fluid sediment. J Neuropathol Exp Neurol. 1970 Apr;29(2):320-30?

I believe this question is more philosophical than practical. However, I can say that while neurodegenerative diseases may exacerbate psychological symptoms and potentially affect how memories are processed or recalled, this would not typically be considered repression in the psychoanalytic sense. In clinical practice, cognitive symptoms coming from neurodegenerative diseases are generally treated and managed differently from issues arising from psychological repression.

The question being reformulated is (if you agree!): In clinical practice, do we neglect the concept of repression when dealing with neurodegenerative diseases?

Hello,

The administration of adeno-associated virus (AAV) vectors via intracerebroventricular (ICV) injection necessitates careful consideration of the vehicle used to ensure optimal delivery, stability, and efficacy of the vector. The choice of the vehicle is crucial as it can significantly influence the distribution, expression, and overall success of the gene therapy.

In conclusion, while PBS and aCSF are commonly used and generally effective vehicles for ICV injection of AAV vectors, the final choice should be guided by the specific requirements of the AAV serotype, the experimental design, and the need to maintain the physiological integrity of the brain tissue.

This list of protocols might help us better address the issue.

The most aggressive model for amyloid plaques is I believe the APPNL-G-F model, which first gets plaques at 2 months. Also the 5xFAD looks to get plaques as early as 2 months.

A great resource for comparing animal models in Alzheimer's disease is available on Alzforum (--> Research models --> Alzheimer's disease).

Maybe the following 2 articles can help to answer your question.

You touched a very tough subject that is penetrating the whole biology and medicine: the hypothesis creation and testing. Simply said, a combination of Complex Systems measures and AI together with machine is a very useful tool that can help to address similar problems to yours.

When you are really interested about complexity measures, which are employing entropy, their application to ECG (EEG is not much different), and subsequent creating and testing of Hypotheses using AI and ML, you can read the methodical paper on prediction of heart arrhythmia. The paper explains all from the first principles and have a rich list of related literature.

After reading it, when you find it useful, it would be possible to continue in our discussion, as the subject is really uneasy to grasp without thinking through it all.

Just one remark, any habits, comorbidities, and other health related features can be easily incorporated into AI & ML methods. I recommend reading papers on gut microbiota health/dysbiosis, which I shared in the project "Complexity in Medicine…" The research is linking gut microbiota dysbiosis to neurodegenerative diseases!

Thanks Farhad Kamandi

Paraphrasing and collecting reference, editing work in other word, cannot result in coautorship ! The authors of papers must have been involved in the creative process of the paper, no ? To be honest, I do not think that this kind of request is deontological or even ethical.

in fly model MPTP can be used to induce parkinsons disease refer the following article for further clarification.

"Resveratrol prolongs lifespan and improves 1-methyl-4-phenyl1,2,3,6-tetrahydropyridine-induced oxidative damage and behavioural deficits in Drosophila melanogaster"

Chinese Medical Journal may be a good choice. It is an authoritative medical journal with a long history in China, and its status is similar to that of BMJ.

Yes Surely

Yaser Rafiq Mir I could not find a video tutorial that explains the process. However, I found this webpage that shows the different step necessary to align sequence traces to a reference sequence.

The reference sequence can be imported from several databases, imported from FASTA files, or pasted in as text.

Hope this helps

I do not think there is a correlation between the quality of the review and the review duration. After one year of review, I may receive a review of low quality; on the other hand, after only one month, I may receive a review of high quality.

"Responsible peer reviews" should consider time during the review process. This is science! And it's being changed on a repaid basis.

Signs of Neurodegeneration in mouse brain include.

1- Distortion of layers and neural organization

2- Cell shrinkage

3- Pyknosis

4- Hypertrophy

5-Reduced Number of neurons

6-Poorly stain neurons

7-Cavitation

There most likely are many causes of AD.

One prominent theory is loss of autophagy. This dysfunction

allows deposition of tau protein. Rapamycin inhibition of the

mTOR pathway allows improvement in autophagy and repair

of damaged proteins etc.and suggests therapeutic effects.

Mitochondrial dysfunction is also involved. The loss of normal

cell function due to severe mitochondrial damage from various insults

ie ischemia, toxins, hyperglycemia, direct physical damage etc.

Cells die at an advance rate via apoptosis.

I love this topic....so much to learn

Lester Mandelker DVM

Fellow AAVPT

looks like IHC works https://www.nature.com/articles/nbt.4281 , i have not used it but i work with IHC for 25 years and i see good IHC data in this paper, try to go through this paper, i think it will help you

Neurofilament light chain.

Robert's questions are very important and his comments about Gosset's t are dead-on, yet I am still not sure of the question. In this case, "multiple comparisons" would typically refer to a situation in which there are multiple contrasts between groups, and the need, therefore, to handle alpha appropriately with that in mind. Yet, from your last post, I think that I see two groups, thus only one contrast.

Now, it seems that you do not view this as repeated measures as you are averaging the three measures; this is for you to decide, and involves your research question more than statistics. I would have no difficulty using a t-test with an N of 10 in each group, unless there is another problem. For instance are there distribution irregularities? I say this because I see a recommendation of a non-parametric test. If it is so simple as heterogeneous variances, use of a Welch-type t-test is a solution.

Finally, if all else fails, you can avoid all statistical distribution theory by making your own distribution with a re-sampling plan such as bootstrapping.

Hi Matvei Khoroshkin , the forerunner in RNA-seq data for AD and dementia and even TBI is the data atlas from Allen Institute which is quite solid and reliable and gives you many options to filter your results:

For PD RNA-seq data check:

You may also check the following website for papers with different RNA-seq data:

Yes. Please take a look at the following RG links.

According to the type of neurodegenerative disease.

1-For neuronal degeneration

presynaptic growth-associated protein B-50 (also known as GAP-43, neuromodulin, F1).It's a marker for mossy fiber sprouting associated with neuronal damage as in hippocampal sclerosis.

2-Multiple sclerosis: myelin stain, luxol fast blue.

3-Axonal injury: ß-amyloid precursor protein.

4-Gliosis: as in multi infarct dementia, GFAP

5-Neuropil and axons are well delineated by silver stains.

6-Microglia by CD68

7-Apoptosis by caspase 3.

The pruchures of these products are available online with the detailed techniques for each.

Trehalose is of particular interest having been shown to reduce protein aggregates in models of other neurodegenerative diseases (Tanaka et al., 2004; Davies et al., 2006). Trehalose is a naturally occurring disaccharide, which appears to play an important role in stress responses in yeast (Singer and Lindquist, 1998). Whilst it has been suggested that its ability to reduce protein aggregation occurs due to a chaperone activity, or through binding and stabilization of abnormal proteins (Tanaka et al., 2004; Davies et al., 2006), it has also been shown to act via an mTOR-independent pathway to increase autophagy (Sarkar et al., 2007; Rodríguez-Navarro et al., 2010). These studies have clearly prompted interest in this compound, though a recent study of mouse primary cortical neurons found that trehalose did not prevent toxicity from exposure to α-synuclein pre-formed fibrils (Redmann et al., 2017).

Mini Review ARTICLE Front. Neurosci., 08 October 2018 | https://doi.org/10.3389/fnins.2018.00693 Emerging Treatment Approaches for Parkinson’s Disease Thomas B. Stoker1,2,3*†, Kelli M. Torsney1,4† and Roger A. Barker1,2,3

Chere Marianne Tous les Vasodilatateurs ,et les fluidifiants sanguins ont des potentialités " limitées cependant dans la réalité clinique avec le temps"

Nos produits ont des actions réellles que nous souhaitons developper en preclinique et clinique avec les moyens qu il faudrait. Nous sommes surs de nos résultats nous attendons le financement ou avec qui travailler .

<< Je ne pourrait pas vous devoiler le comment cela marche ,mais il ya des similitudes avec calcium channel blocker et aussi avec le systeme autophagique et qq chose d autre que nous gardons en secret )<< jusqu a l étape d une convention de travail et de partenariat .

Hi researcher,

You can use the lipid-based nano-biopolymers that functionalized with targeting agents such as monosaccharides, antibodies, folic acid, etc. in which smart nanocarrier actively target the mitochondria membrane.

Simply put, when the neurons start to die away, this will eventually lead to loss of neuronal connections and circuits. Then, this would obviously cause decrease of functional activities or shrinkage of certain brain regions, which may be termed as 'neurodegeneration'. I hope this answer helps.

I hope you find it useful

Digit Cancellation Test:

Zazzo R. Test des deux barrages. Actualités pédagogiques et psychologiques . Neuchâtel, Switzerland: Delachaux et Nestlé; 1974.

Pattern Comparison Test:

Salthouse TA Babcock RL. Decomposing adult age differences in working memory. Development Psychol . 1991;27:763–776.

Regards

We conducted few experiments, which were more like an extension of our major project to answer the above question. Anybody interested can look into our publication https://www.researchgate.net/publication/309034150_Admixing_of_MPTP-Resistant_and_Susceptible_Mice_Strains_Augments_Nigrostriatal_Neuronal_Correlates_to_Resist_MPTP-Induced_Neurodegeneration?_sg=SduYxo0hz3DQmqy93kk9hyMjV_J-i0ouvHaEbuqxedt9oNHu5fli5E-5pZcgV85KPj4wkgogp_LGPrjwEADHJUw-jqh2GloUOcjthiGF.f0rENiJvOQ7gnlJA4_ETEjlNMaOTd1ddCDFJh6nABeNCD5Znzuxw_90ArlhGmSd80SC3nlE_R50L_Iw3FVWmnQ

Mohamed Mahdy Thank you very much for the links! The last link isn't working, could you giev me the title of the paper?

Thank you. That is helpful--I will have to take a closer look at the GATA-1 paper. I also found a paper by some UK researchers in 2002 (Flynn, PMID: ) which found CCR3 on microglia to be upregulated by TNF-alpha and interferon-gamma. CCR3 on astrocytes appeared to be upregulated by TNF-alpha, but not by IFN-gamma.

Thank you very much Mr Abdelrahman Ibrahim and Mr Rafael Linden.

Finally, a Winner for Alzheimer's? Anti-amyloid Agent Shows Promise

Pauline Anderson

July 26, 2018

CHICAGO — Positive results for an anti-amyloid agent in patients with early-stage Alzheimer's disease (AD) is drawing praise, but experts are calling for caution.

Results from the new phase 2 study showed a statistically significant reduction in brain amyloid with a high dose of BAN2401 (Eisai Co. Ltd/Biogen Inc) at 18 months.

In addition, the study showed a dose-dependent, statistically significant, and clinically meaningful slower decline in cognition and function with the highest dose compared to placebo.

"This is the first large clinical trial to support the amyloid hypothesis," Lynn D. Kramer, MD, chief clinical officer and chief medical officer, Neurology Business Group, Eisai Co, Inc, told a press briefing.

The study was released here at the Alzheimer's Association International Conference (AAIC) 2018.

The multicenter study included 856 patients with early AD (mild cognitive impairment due to AD or mild AD dementia) and amyloid pathology confirmed by positron-emission tomography (PET) or cerebral spinal fluid (CSF) tracer.

At baseline, the mean Mini‒Mental State Examination score was 25.6; the mean Clinical Dementia Rating Scale‒Sum of Boxes (CDR-SB) score was 2.9, and the mean Alzheimer's Disease Assessment Scale–Cognitive Subscale (ADAS-Cog) score was 22.2.

Ideally both PCR and WB, but if you have budget for only one the latter makes more sense especially if you plan on publishing. For PCR validation, you could randomly choose several genes that are not differentially expressed and some genes that are differentially expressed. Genes of interested could be in the pathways that are pertinent to your investigation i.e. inflammation. I will give you an example of how my previous PI represented microarray results to give you an idea.

Is there a specific question regarding sciatic nerve compression you might want to ask?

Look, it's a good question ... I try to give a ontological answer but also (above all) a functional one and, in the end, it is the applied chemical stoichiometry that will tell us if budgets exist that do not return and in those mysterious areas we must go to investigate. Chemistry will not support the needs of dialectics even if expressed by authoritative scientists. Truth wins in science, and it is evident that chemical stoichiometry makes this research more fruitful.

The cell body of the neuron needs a lot of energy and the mitochondria can not provide everything that is needed. However, the mitochondria contained in the cell body are very few and this is just a conundrum: in fact it is paradoxical that a cell that consumes a lot of energy with a request for high oxygen supply for biological combustion has few mitochondria!

Different facts lead us to think that, thanks to the probable fusion of the mitochondria with the Nissl substance (Nissl: the endoplasmic reticulum of the neuron-soma) this gives the soma the “machine” that synthesizes all the ATP it needs but… in the very long axon that happens? The endoplasmic reticulum does not penetrates the axon and it’s true-like that myelin support this energy demand, especially for firing (see " Monitoring ATP dynamics in electrically active white matter tracts” Elife. 2017 - PMID: 28414271) let us not forget, is energetically dispersive. So the white matter, but also the gray matter, energetically supports the axon with myelin. Among other things, there is evidence that neural activity stimulates myelin growth. Keep in mind that the brain has a serious deficit, compared for example to the muscle: it does not possess myoglobin which is slow reserve of oxygen so much that it is extremely affected by ischemia, while for the muscle this criticality does not exist, thanks to myoglobin.

So myelin is home of an active energy metabolism with aerobic degradation of glucose, which, through the Krebs cycle - which we have shown to be very active in myelin (see our article “Tricarboxylic acid cycle-sustained oxidative phosphorylation in isolated myelin vesicles” PMID: 23851157), supports "outside" the neuron's energy needs, above all to support firing.

But there's more!

In fact, for go on the combustion the myelin need a lot of oxygen and that's why the myelin has the highest content of carbonic anhydrase (CA) of the whole organism. Let us remember that CA is the enzyme that combines the carbon dioxide produced in gaseous form with water to make it soluble in the form of bicarbonate. Remember us that if the CO2 remained gaseous (let's remember that in this state it is produced by the Krebs cycle) in the brain we would have terrible seizures! Furthermore, again to be in the subject of oxygen, myelin dissolves oxygen quite well and therefore constitutes a slow reserve of oxygen to support neuronal combustion, even if it is not as efficient as myoglobin. In fact, white matter resists better the hypoxia of gray matter.

So many data would then remain mysterious if we do not take into consideration the chemical reactions we have now mentioned are operative in myelin.

At last to answer with the ontological profile, I hope that I have answered your question, and we can realize that all this chemical support to the axon from myelin could not be supported by the neuronal cell alone. I believe that it is possible to look at the issue from a teleological point of view, but if we talk about it we will talk about it another time…

Yours sincerely,

Sandro

‘Saltatory conduction’ is actually a nickname. Action potentials do not ‘jump’, nor do they even ‘travel’ along an axon, myelinated or not. The term action potential refers to a measurable fluctuation of the potential difference between an intracelular and an extracellular electrode, which  arises as a consequence of ionic fluxes across plasma membrane channels. Each action potential is a local event generated at a very small patch of the membrane, where there is a high concentration of a particular type of sodium channel. The source of the rapid depolarization known as the action potential is actually the massive influx of sodium ions through such voltage-gated sodium channels. These channels are more or less evenly distributed along the plasma membrane of unmyelinated axons, but they are for the most part restricted to the Ranvier nodes in myelinated axons. The fact that individual action potentials in a normal myelinated axon are generated at succesive Ranvier nodes leads to the appearance of a saltatory phenomenon. In reality, however, each spike is a single event. What drives the conduction of action potentials along the axon are electrotonic currents between the segment of the axon where an action potential has just been generated, and the rest of the axon ahead of that region, along the longitudinal dimension of the fiber (forward intracellularly, backward extracellularly). Myelin helps focus such currents into each node of Ranvier not quite because of its electrical resistance, but mostly because of the low capacitance of the myelin stack, which drains very little current through the membrane and leaves most of those local currents to spread longitudinally and sink into the concentrated voltage-gated sodium channels at the next Ranvier node. Then and there, the current drives the opening of those channels, a brand new action potential is generated, and so on.

Hi,

I think you might find this helpful

All the best !

Hi Mark Gordon,

Thank you for your response.

Currently, the only known modifiable risk factor for glaucoma is intraocular pressure(IOP) and hence is the focus of most physicians for glaucoma treatment.

But I understand why you ask this question as there is a paradigm shift in our understanding of glaucoma over the past 5 years or so. Newer drug classes like the ROCK kinase inhibitors, adenosine receptor agonists and newer prostanoid receptor agonists are also on the brink of entering markets after successful trials. Advanced modalities like SiRNA and Gene therapy has also been explored for glaucoma treatment.

What is important to note is the need for this research as glaucoma progression has been noted in several patients despite being maintained under target IOP and also invariably in NTG patients. The role of mematine, brimonidine and other neuroprotective agents has also been explored but results don't match up to the expectations. There is a lot to do yet but at the same time IOP is something that is imperative to control. These anti glaucoma drugs thus still have a very important, though quickly diminishing importance in explaining the patho-physiology of glaucoma, which unfortunately yet excitingly remains an elusive hunt for glaucoma researchers to pursue.

An excellent review of the newer drugs is attached for your reference.

To help facilitate attachment, cell spreading, growth, morphology, differentiation PDL or PLL are used. In histochemical applications both polymers of D- and L-lysine are used to coat slides to promote attachment of cells. i use astrocytes cell for cell viability test in 96 wells. i found my cells are easily attached to the cell well without PDL or PLL coating.

may be useful

Hi I am sending EtBr/AO staining protocol  used in my lab

First make cell suspension in PBS.

Now mixed the cell suspension with EtBr/AO staining solution (1 mg/ml of AO in PBS + 1 mg/ml of EtBr in PBS) and incubate for 2 minute on ice.

Now mount the mixture on slide and visualize the cells under fluorescence microscope.

Capture the images using TRITC,FITC and & DIC. After merging the image using any software u can differentiate the apoptotic cells.   

Hi,

are you restricted to LDH as endpoint for cytotoxicity? Remember that it measures membrane integrity, which may still be uncompromized following the 24h treatment with MPP+, which interferes with energy metabolism. You might want to try a different cytotoxicity test with an endpoint more closely related to your substance's mode of action, e.g. MTT (simplified measuring cellular respiration), Neutral red uptake (indirectly influenced by ATP contents) or direct ATP measurement, which in my opinion would be the most appropriate approach for your substance.

Best regards

Thank you Spielman and Kondru for your contirbutions,

highly appreciated

HI Igor! Nice to hear from you. I have a vast library of electrophysiology, microperimetry, mesopic visual fields and VA, mainly in non-DR (AMD, PR, Stargardts, etc).

What we see is:

1. SDM improves electrophysiology in every disease

2. The nature of the electrophysiology improvement is different and characteristic for each disease, as "reset" theory predicts.

3. Mesopic VA and perimetry improves in every disease.

4. By linear regression, the worse the pre-treatment findings for any measure, the more the improvement after SDM. Therefore, what we're seeing is restored function to previously non-functional (presumably pre-apoptotic) tissue (rescue). This is where I would expect DARC to show a reduction / disappearance of pre-apoptotic cells. 

Best,

Jeff

Dear Ms. Correia,

Is there any cheaper way to do this?

Thanks in advance,

Alam

Stress to endoplasmic reticulum (ER) can cause UPR and aggresome formation. Aggressome accumulation has been shown in AD. The ER stress is also related to autophagy. So theoretically autophagy can be a target for AD treatment. The thing is when you start to study a new signaling pathway in a disease there is always some association. The issue becomes how to decipher your findings.

@ Dr Dick Terwel. l am grateful for the pdf you sent. Thanks for your prompt response to my request.

Hi Lien!

Neurodegeneration is a multistage process, which depends on various factors. First of all, you have to decide what a model you want to use and what kind of cellular changes you want to see. Neurodegeneration may start from peripheral parts of neuron or may be initiated in the body. One more important question is how long it takes to develop neurodegenerative changes. When you will be able to answer these questions for yourself you can start to choose appropriate IHC or IF methods of detection.  

Thanks Dr Serra-Mestres,

This is exactly the information I am looking for. Now, it would be easier for me to do the literature survey easily.

Thank you so much.

Regards, Dushyant

Yeah.. There is one paper from our research group which shows mamba venom inhibits the aggregation of amyloid beta, but still the BBB is a mystery.

Dear Sweilem:

This important link for you

Wells, J. D., S. Thomsen, P. Whitaker, E. D. Jansen, C. C. Kao, P. E. Konrad and A. Mahadevan-Jansen (2007). "Optically mediated nerve stimulation: Identification of injury thresholds." Lasers Surg Med 39(6): 513-526.

uses "hematoxylin and eosin (H&E) to demonstrate pathologic changes in the tissue. Selected unstained slides were submitted for Luxol fast blue-Periodic acid Schiff stains (LFB-PAS) to highlight myelin and collagenous connective tissues. "  "thermal injury, denoted by birefringence loss, can be detected in H&E-stained sections, the resolution of subtle changes is significantly improved when sections are stained with dyes that enhance the tissues’ natural birefringence. For example, picrosirius red (PSR) enhances collagen’s birefringence and has been shown to enable detection of thermal injury in collagen and muscle. "

Dear Xinjian,

Though you have not been specific on your research population (human/animal) or how you model your CPP ( Three-compartment chamber (or 2 or 4), and how are they defined; do you have a "forced answer"), I suggest reading the following chapter: https://www.ncbi.nlm.nih.gov/books/NBK5229/

Please pay attention to section 4.6.2. Psychostimulants, as it might have some suggestions on why you fail to get  excitation effect.

Hope this helps.

Success!

Jelena

Hi Mathias,have a look to these two papers. Maybe it can help you

Hello, you may check this artcile and if this model of tauopathy in rats fits your experiment, you can contact dr. Michal Novak or dr. Peter Filipcik for more information. This model is used for a long time and is described very well, but I am not sure if it is provided only to collaborators or also commercially distributed. Good luck

For tau soluble oligomers, you could check these recent papers by Marc Diamond and Brad Hyman's groups.

PMID 24857020, PMID 27974162, PMID 26458742, PMID 27351289

These are pubmed ID numbers.

You can also use murine models of TDP-43 overexpressed in cortical neurons

I am interested in learning more about how I can help. I am using Drosophila as animal model to study dopamine related diseases and I am in the process of acquiring/making innovative genetic tools that could be useful to help you answer the question, in vivo. Looking forward to your response.

Best,

Dr. Tito

HIV is associated with difficulties in many cognitive domains, including short term memory which eventually gets consolidated into long term memory and learning.

A full screen for cognitive functions is essential for all HIV patients, especially those that can expose themselves or others to harm from cognitive impairment.

I could not agree more with Prof. Onaolapo, but again it all depends on your sample size. MANOVAs with age and APOE genotypes as covariates may be acceptable.

Dear Adejoke,

Hope these papers are useful.

DOI: http://dx.doi.org.uml.idm.oclc.org/10.1210/edrv-1-2-167

Thank you all. Farid is particularly acknowledged

Hi

I would use colour segmentation plugin in imagej/fiji

you can eventually preprocess the image with colour deconvolution plugin

of course white balance is very important during photo acquisition.

Hope this will be useful.

Regs

Carlo

Ha, think those are a little bit too big for endocytosis? ;)

1. The *very* nice thing about these beads is that they do not fill the processes: the signal is only in the soma, and is punctate (probably clumps/vesicles filled with many beads). With reasonably good imaging, you can readily distinguish between "filled" red cells like you'd get from RV, and bead-labeled cell bodies. Have a look at the (Kiritani et al) paper that I attached - this is what Taro did: RV-GFP together with two different colors of beads; I think he has a figure where you can see the difference in labeling pattern.

2. Have a look at the retrobead spectra here, if you like: https://lumafluor.com/Information.php

3. The other option is to go with cholera toxin subunit B (CTB), conjugated to a dye, for instance one of the Alexa variants. You can get quite a range. I've used CTB-Alexa-647 when I already have red and green dyes in the tissue. Of course, you'll need an imaging system that can detect far red. I believe you can get CTB conjugated to other dye families too. This will give you a cytosolic signal, unlike the Lumafluor beads. As I recall, in the (Kiritani et al) paper, Taro also used CTB together with RV. 

I can't say whether either of these options is better than Fast Blue, but at least they've been used (and published) in conjunction with RV.

Your protocol looks OK - we used prefused but not paraffin-embedded tissue (mouse brain)  but otherwise our procedure was just like yours. However, I'm not sure if FJ has been used in zebrafish already - do you know about such a publication?  As I am a mouse person, I unfortunately cannot offer specific ideas about possible reasons this dye might behave differently.

Go to: MBL weg servic, mouse atlases: mbl.org/mbt_main/atlas.html.

Making your own P21 mouse atlas in 10 steps. Succes. E.Marani

Hi Touqeer,

Some of the most expensive components of an in vivo and in vitro setup is the anti vibration table. You can avoid buying such system by putting a table, prefereabily a metal one and just the square stuff, over an old big truck tire or over four half tennis balls in the corners. The metal will help because most of the micromanipulator bases can be attached to the table with a magnet, helping to stability and giving flexibility in location. The table should weight enough to provide stability and bend a bit the rubber. This option is used in labs even for single-cell patch clamp recording and should provide stable recording conditions for extracellular field potentials both in vivo and in vitro.

Hope these solutions help!

Kind regards,

Marcel 

Docking programmes are very useful to detect the inhibition potential of a ligand (small molecule) against bulky molecule (protein). They can tell about the specific area and type of bonds could make and it's stability. These results usually similar to the in vitro studies. Auto dock docking programme is available and free to use. With expert advices and few practices valuable information can be gained.

Hi Nielsen

Are your sections one set of serial sections? Do these 5 sections cover from the beginning till the end of the lesion?

If this is the case you could do the following to estimate the lesion volume. Use Image J software  to measure the lesioned area in each section (if you calibrate it as Amit suggested, you will get mm2 values). 

Then calculate a lesioned average [LA]  for each animal, and ESTIMATE  average volume  by multiplying the LA by the section’s width (0,02 mm), and the total number of sections [TS] obtained for the whole lesion (that is to say, if you collected 6 series of 5 sections each, your TS would be [5 sections x 6 series]-5 sections {because you don't count the five sections after your last section} = 25).

Another similar way of going about is, is (using the same numbers as above) to consider the avarage area between Section1 and section2, and ESTIMATE that the lesion volume in the 5 sections between those is that average [...] and do the same for each set of sections.

Hope it helps!

I am not an expert of Friedrich ataxia. As Friedriech ataxia is a neurological disease in nature, a neuronal culture should be employed theoretically at best. However, fibroblasts from these patients may also be useful.

At the top of my head I don't recall a specific paper that looked into this correlation (or lack thereof), but it's not my area of specialization. Hence, I would suggest a thorough PubMed search, which may turn up some papers that deal with this question.

I do not disagree with the opinions above, but motor behavior and weight can vary significantly even in inbred strains. Although some favor inbred strains for genetic homogeneity, it may also make results less generalizable and can occasionally produce unusual GxE interactions.. This is why we do clinical trials in humans with diverse genetic backgrounds.  When backcrossing, you may also find that fertility or pup survival suffer greatly.

Aging and BBB dysfunction is an interesting topic because its a high risk factor for developing neurodegenerative diseases. To my knowledge most of recent studies point at loss of tight junction proteins of the BBB with aging which is likely due to neuroinflammation. Infiltration of circulatory leukocytes and blood borne immunoglobines into the brain is a marker of BBB leakage that happens in aged humans. In aged mice however, only loss of TJ proteins of BBB was observed. I think its not clear which come first, neuroinflammation or BBB leakage. It could be systemic inflammation disrupts the BBB first and lead to infiltration of toxins to the brain that trigger neuroinflammation. Not sure if the brain itself with aging would autonomously initiate inflammation due to certain malfunctioning in metabolic processes and clearance pathways to remove waste byproducts !?.. I would love to here comments from experts in the field..

I am a Sr Administrative Assistant with no knowledge of this at all.

Quantification of immunohistochemistry depends on a number of parameters including what the distribution of the staining looks like for each antigen. Is it somal (cell body), neuritic, puntca-like, etc? Depending on this you have many choices for quantification. Are you looking to see if all three labels are in the same cell or same field, if their intensities vary, etc? The choice of how one quantifies is dictated by what the staining looks like, and what you are trying to achieve. I've used ImageJ, Imaris, and in-house methods to quantify- each used for very specific types of staining. I also cannot stress enough that your staining needs to as good as you can possibly get for accurate measures.  We probably need more information to be helpful to you and  why Image J wasn't a