Neurobiology - Science topic

Dear Ranjit Singha , I am not an expert in this topic, but, as someone who deals with AI and neural networks, I may recommend you four articles that may be very related to spreading and communicating the emotions.

"Researchers have charted a long-distance ‘wireless’ nerve network in Caenorhabditis elegans worms for the first time. The nervous system can be thought of as a web of neurons that pass on messages through direct links, called synapses. But neurons can also communicate over longer distances by releasing molecules called neuropeptides, which are intercepted by other neurons some distance away. Incorporating both ‘wired’ synaptic connections and wireless signalling better predicts how signals travel in the worm than does a model using synaptic connections alone...

The next steps for Schafer and his colleagues will be to undertake similar studies in other organisms — aiming to understand how the neuropeptide network, in combination with the ‘wired’ synaptic network, contributes to an organism’s behaviour. A technique published in Science last week that allows researchers to visualize where neuropeptides bind to their receptors could aid in this quest. Because neuropeptides are conserved across species, some researchers suspect that this network could look similar to those in other organisms, including humans..."

Artificial intelligence (AI) has become an integral part of various fields, including neuroscience and mental health research. In recent years, AI has significantly contributed to the ongoing research and understanding of the neurobiological factors associated with depression. By analyzing vast amounts of data, AI algorithms can identify patterns and correlations that humans may overlook. I will discuss how AI is revolutionizing depression research by enhancing diagnosis accuracy, predicting treatment outcomes, and uncovering novel insights.

Firstly, AI algorithms have shown remarkable accuracy in diagnosing depression based on neurobiological markers. Traditional diagnostic methods heavily rely on subjective assessments by clinicians, which can be prone to biases and errors. However, AI systems can analyze brain imaging scans and genetic data to identify specific biomarkers associated with depression. By comparing these patterns with a large database of known cases, AI algorithms can provide more objective diagnoses with higher precision.

Moreover, AI has the potential to predict treatment outcomes for individuals suffering from depression. By analyzing a patient's neurobiological data alongside their response to different treatments, AI algorithms can identify which interventions are most likely to be effective for specific individuals. This personalized approach not only saves time but also increases the chances of successful treatment outcomes.

Furthermore, AI is capable of uncovering novel insights into the complex neurobiology underlying depression. With its ability to process massive amounts of data from various sources simultaneously, AI algorithms can detect subtle patterns that may go unnoticed by human researchers. These discoveries could lead to breakthroughs in understanding the underlying mechanisms of depression and developing more targeted treatments.

In conclusion, artificial intelligence plays a crucial role in advancing our understanding of the neurobiological factors associated with depression. Through its ability to analyze vast amounts of data quickly and accurately, it enhances diagnosis accuracy while predicting treatment outcomes for individuals suffering from this debilitating condition. Additionally, it uncovers new insights into the complex nature of depression that could lead to improved treatments in the future. As technology continues to advance, AI will undoubtedly continue to contribute significantly to depression research and ultimately improve the lives of those affected by this mental health disorder.

Yes. I did this for two years.

A Sutter P97 or a DMZ electrode puller offers more adjustment options, but with a little testing you will get good patch electrodes with the PC100 too.

Psychobiotics for Mitigation of Neuro‐Degenerative Diseases: Recent Advancements - Dhyani - Molecular Nutrition & Food Research - Wiley Online Library

Hi Shaktiprasad, I am the Product Manager of the neural team at STEMCELL Technologies. We need a little more information and context for your question. Please contact our Product and Scientific Support team at techsupport@stemcell.com and a team member will troubleshoot with you.

Kind regards, Nusrat

Can you also forward the drive and software to me. My email is sargol.okhovatian@mail.utoronto.ca

David Crompton , David Martínez-Vargas Janelle M Fine and

Dear professor,

sorry but I have many problems to partecipate at the Conference because of my cronic heath problems.

All my best, Catina Feresin

Vahid Rakhshan

I apologize for any confusion my previous response may have caused. Let me clarify:

There is evidence that some synapses are stronger than others, and that synaptic strength can change over time through a process known as synaptic plasticity, which includes long-term potentiation (LTP) and long-term depression (LTD). These changes in synaptic strength are thought to be important for learning and memory.

However, it is not clear that any single axon-branch to dendrite mini-synapse involved in memory formation is larger or stronger than any other one in general. The strength of a synapse is determined by a variety of factors, including the amount of neurotransmitter released, the number and sensitivity of receptors on the postsynaptic membrane, and the properties of the presynaptic and postsynaptic neurons themselves. These factors can vary widely from synapse to synapse, even within the same neural circuit.

So, while there may be differences in synaptic strength between individual synapses, it is not accurate to say that any single synapse is always larger or stronger than any other one, or that the strength of a synapse is solely determined by its size. The idea of synaptic weighting and LTP/LTD remains an important and valid concept in the field of neuroscience.

On the other hand, I am also, among other things, a philosopher, a clinical psychologist and a theoretical physicist. I tend to look at commonly accepted definitions and paradigms from many different perspectives.

I worked as a neuroscientist for 2 years at the Medical University of Wrocław, I previously studied neuroscience at Duke University. I did not find answers to my questions. On the one hand, we are looking for something, while on the other hand, our accepted definitions and paradigms often lead us to contradictions.

What interests me are noumena (things in themselves). I look for them in everything, although they can be a difficulty in typically technical discussions. Nevertheless - I cannot believe in any theory, concept or description if I do not find assumptions deep enough in them. The foundations are often fragile.

Neuroscience is one of my favourite disciplines of knowledge, but I nevertheless think that its foundations are not clear enough. I think the same about theoretical physics.

Hence, I warn you that what I write may be risky at times, but I take part in this discussion because I myself very much want to understand and be able to reflect further.

The type of memory you are describing is often referred to as highly superior autobiographical memory (HSAM) or hyperthymesia. Individuals with HSAM have the ability to recall detailed and specific information about events from their own life, as well as information from other domains, with exceptional accuracy and vividness. They often report that these memories come to mind involuntarily, without any conscious effort to retrieve them.

The inability to update or revise memories in individuals with HSAM may be related to the way their memories are stored and retrieved. Research suggests that individuals with HSAM have highly efficient and reliable retrieval processes, which allow them to access memories quickly and accurately. However, this efficiency may come at the cost of flexibility and updating, as memories may become so strongly encoded that they are resistant to change.

The causes and mechanisms underlying HSAM are not yet fully understood. Some research has suggested that HSAM may be related to differences in brain structure or function, such as increased connectivity between certain brain regions involved in memory processing. However, more research is needed to fully understand the underlying neural mechanisms of HSAM.

It's important to note that HSAM is a rare phenomenon, and not everyone with exceptional memory abilities has HSAM. While memory abilities can be enhanced through various techniques and strategies, the type of memory observed in individuals with HSAM appears to be a unique and innate ability.

I know a bit more about this from autopsy as well. I was also diagnosed with autism spectrum disorder as a child.

Vahid Rakhshan I will definitely spread the word about this amazing initiative. It's great to know that we can contribute to such a noble cause by simply utilizing our excess computer power. Thank you for bringing this opportunity to my attention. Let's join hands in making a difference in the fight against diseases.

Dear Colleague Michael Marek,

Yes, this is so often the case. My husband who is an active observational planetary scientist says how often the "devil" is in the details of data analysis.

Our short story collection, Children of Steel, is being considered by Wayne State UP, BTW. It is a collection of short fiction by people who grew up in steel mill towns. I just noticed where you retired from teaching.

Hi Mohab,

You could have a look at the NEURONET Knowledge Base

It provides a summary overview of the Innovative Medicines Initiative neurodegeneration research portfolio through an interactive dashboard, the NEURONET Knowledge Base includes links to over 500 publications and more than 380 publishable deliverable reports, acting as a one-stop shop to explore the diverse projects and outputs of the portfolio.

On the project website you can also find an overview of the ongoing (also completed) public-private research projects in the area of neurodegeneration (including dementia & Alzheimer's disease): https://www.imi-neuronet.org/ongoing-projects/

Hope this helps a bit.

Best wishes,

Chris

Check https://sitn.hms.harvard.edu/flash/2019/exploring-the-underground-network-of-trees-the-nervous-system-of-the-forest/

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"

Dear Alejandro!

Please You look at the following protocol:

NSC-34 cells were cultured in differentiation medium consisting of minimum essential medium Eagle/alpha modification (Millipore-Sigma, Burlington, MA, USA) supplemented with 1% fetal bovine serum (Thermo-Fisher Scientific, Cambridge, MA, USA), 1% 100× MEM non-essential amino acid solution (Millipore-Sigma, Burlington, MA, USA), 1% pen strep (Thermo-Fisher Scientific, Cambridge, MA, USA). D

Shweta Singh Fantastic. Thank you very much. We are asking (neuro)psychologists, neurosurgeons, speech-language therapists, occupational therapists, physiotherapists, and psychotherapists to participate. If you have a few contacts you would like to share, please message me privately. We will be happy to reach out to these people directly, if it helps save your time. I am also providing my email address: MPolczynska@mendet.ucla.edu just in case. Best wishes, Monika

Everyone who is listed as an author should have made a substantial, direct, intellectual contribution to the work. For example (in the case of a research report) they should have contributed to the conception, design, analysis and/or interpretation of data. Honorary or guest authorship is not acceptable. https://hms.harvard.edu/sites/default/files/assets/Sites/Ombuds/files/AUTHORSHIP%20GUIDELINES.pdf

Compared to various living organisms characterized by a much lower level of organization and body structure, the human body has limited regenerative abilities. However, along with technological advances, in medicine, genetics, microbiological tests, etc., the possibilities of transplanting various organs, limbs, growing specific types of tissues and rebuilding certain parts of the human body are gradually increasing. One of the most difficult and perhaps impossible to implement in the future is the rebuilding of the central nervous system, including the human brain. Similarly, it will be extremely difficult in the future to build artificial awareness in artificial neural network systems as a continuation of the progress made in the development of artificial intelligence.

Best regards,

Dariusz Prokopowicz

When AI began in 1956 (Shannon McCarthy, “Autmata Studies”, 1956), there was no connection between AI and neurobiology. The two most influential people were Marvin Minsky and John McCarthy. Minsky said “solve the problem, ignore neurobiology”, while McCarthy believed everything could be solved by using formal logic. In 1956 the founders believed the problem of language would be solved in 10 years! (Still not, in 2021) In 1969 Minsky and Papert published a book where they proved that the perceptron could not learn to decide if a pattern was connected.

This effectively killed the neural network- like approach for the next thirteen years, until 1982 when John Hopfield showed that a recurrent network of 2-state neurons could implement content-addressable memory. AI became, roughly, theory of neural networks, influenced by neurobiology. Neuroscience continues to develop with essentially no influence from AI. Today, “deep learning” is the frontier of AI, with remarkable success, but its’ relationship to neuroscience is unclear. Today, it may be that the grid cell and place cells of neurobiology influence robotic spatial navigation. Phew! I hope this sketch is useful. On a personal level, as a graduate student in the 1970s, I was forbidden to use the words neuron or neural network in an AI thesis.

Principles of Neurobiology, Second Edition presents the major concepts of neuroscience with an emphasis on how we know what we know. The text is organized around a series of key experiments to illustrate how scientific progress is made and helps upper-level undergraduate and graduate students discover the relevant primary literature.

you can download the preview version from the given link:

https://s3-euw1-ap-pe-df-pch-content-store-p.s3.amazonaws.com/9781003053972/e937dc89-0405-4bf1-89b8-33ed62142993/preview.pdf?AWSAccessKeyId=ASIAQFVOSJ5747IEEVII&Expires=1615405273&Signature=Sow50ZCxLEwMvgTprKyTZ2nZSfw%3D&response-content-disposition=attachment%3B%20filename%3D%229781003053972_previewpdf.pdf%22&x-amz-security-token=IQoJb3JpZ2luX2VjEFAaCWV1LXdlc3QtMSJHMEUCIDDjRFeswZq7EQe1AUG3D2u3e%2FMCYasl9l06HNZ%2Bj2CNAiEA08GM6bywJa7qvjzf7QG2c01NVqMjMT%2Bt5vq8c%2BXx%2B%2BMqtAMIeBACGgwwMTIxNzcyNjQ1MTEiDE%2FRUQhPw0C%2BWW9D4CqRA4P9HMFJcidDQ1WY9fdq%2FD0%2FGPOFv02tnngGcxU2zN8o6kh2IUszPkGRo5zIjXIJ2zelmMqXV81wxPN4ASe%2FydfRKShPXyuP9ukxG7Lpj90hNWn2sSJc6qDoGXqDHAlTLG9kmoFcrNDljzuw24i%2BqzDP13y95IpRPl7B7x2VCBIBz5%2F6u4QX%2FHxAK6Ud%2By9ufQ2ETF%2BwPOPisvZi9WuDTUUWHqbrH4uN5H7c2ckkGY1EIKaOP8yWqXX7s7Mv7H14laSXzbqbCPWTIEJ893ZvNAXQausqtsv8uBET%2Fwc1NCvlED1ZxzWJfQUgmdGOVDGmkNaXWSPGOthdfLwwUFX%2B5s15rPXeX32%2Fhcm8WTBCpqDITikHFlWF5moLuAeE1ucZrnWsMDYeJcH5SKIHGfm26fv6Fieh5GkbBTUMOVqm7pBo86n3hSgJzB21P5ssi52ksdmfEfuPn7Ufiiugv1PfP...

May I ask if you continue your experiment and find a good agent? I am also thinking about fixing my cells on a coverglass right after treating with Fura-2 AM and I am wondering if fixation processes by 4% PFA may effect the membrane permeability and cause Ca2+ fluorescence to be lost.

Something like this for action potentials ?

Hi Monika,

For me ensembl.org is the easiest. For example COMT

1. you search for the human gene

2. Open the gene file:

3. Select one of the longest and best annotated transcripts

4. Select cDNA in the left panel

If you don't see the SNPs go to Configure the Page and select Variants. If you prefer, you can visualize the SNPs as well in the gene view.

dear definitely many others apps there,but i suggest according user friendly convenience

Request the pertinent permission from its Author or Authors and then make it reliable, validate it, etc. Pisometrically in its new modification and with its own "ad hoc" Normative Groups.

Similar to above. Jaak

Panksepp?

From affect/psychodynamic perspective Phil Mollon

The self is a complex interaction between memory and attention. In this case memory is facts we know about ourselves (self-knowledge), schematic structures built from our socio-cultural context and autobiographical memory. The Self is how we appraise, use, and prioritize not only information in memory, but also various interests and desires. These actions on memory, interests, and desires require varying degrees of attentional resources. Carolyn Jennings makes this point much better than I in her essay (https://aeon.co/essays/what-is-the-self-if-not-that-which-pays-attention). In sum, there is a bidirectional relationship between our Self and how we appraise, use, and prioritize what’s stored away in memory structures as well as our management of our interests and desires.

sorry not

Andrew Chao no the project we were working on this for is currently stalled but will likely pick back up come May/June so if you figure this out it would be greatly appreciated. Likewise if we figure it out I'll send what ever we end up doing over your way.

Monika - the 'general' guidelines are similar to a 'primary' research paper in that the headings are similar i.e. Intro/background, methods (this may include an existing theoretical framework [as methodology] that you used to underpin the model and your literature search and appraisal strategy [if conducted]. It obviously doesn't include ethics, sampling, data collection/analysis.), limitations, further research, discussion, implications for practice, and conclusion/summary.

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:

I was also asking myself your question and then I found

not sure if it is still important to you but it may be for others...

Please take a look at the following PDF attachments.

It is only when you are working with rodents you have to abstain from using females because their estrus cycle disturbs the results.

Hello I am not an expert on that so pardon but it would seem to be on the order of one or a few photons depending on your theory and what fluctuations you speculate matter. Either you are looking for quantum level changes which you could capture with a total surround of the brain or a partial surround with compensatory assumptions, or you are looking for much more macroscopic change such as heat flow, where the resolution would of course be rather coarse particularly with probable difficulties arranging for a tightly controlled experiment.

Put another way, with less confidence you can make assumptions that make the test easier and at least reduce the hypothesis set thereby.

Oh, the phase-locking between spikes and theta LFP change with the moving speed. I get it, thank you!

Can one use N2a cells s.c. injection to study tumor growth in other mice strain then AJ?!

Journal of Visualized Experiments (J Vis Exp, JoVE)

www.jove.com

Hi Marie

You could try AAV-DJ, which is the most efficient serotype in infecting in vitro cells in our experience. Paradoxically, AAV-DJ is not good in tissue infection in vivo.

Genemedi is experienced in AAV production, you could find more information and manual files on this website: www.genemedi.net/i/aav-packaging

And here are more titles:

and of course

There are so many other, just let me know if you need more. Good luck!

Christina

Tanisha Singh,

Thank you so much for your answer and advice.

آسف السؤال ليس من اختصاصي

BioRender might be a good option (https://biorender.io/)

It's an online app that contains a library of pre-made cells, proteins, membrane shapes, organs, lab equipment, etc. that you can drag-and-drop so you don't have to spend time drawing each element of the figure yourself. Saves a LOT of time and the final figures you create are professional-looking. Plus, it's free for educational use.

Just like environmental factors, psychological factors may have an impact on gene expression and its regulatory mechanisms but the extent of impact would be depndent on multifactorial aspects and would need to be ascertained in a case to case basis.

this link is useful

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4445645/

regards

Dear Monika,

Results in some recent meta-analyses (e.g., by Shaofeng Li) have shown that executive control plays a more significant role in explicit/formal than in implicit/naturalistic L2 learning (SLA). Regarding implicit and explicit brain networks, a similar pattern can be detected, e.g., in the paper by Jing Yang & Ping Li (2012). Brain networks of explicit and implicit learning. PloS One, 7(8), e42993.

Hope this above information helps a bit,

Best,

Edward

It seems similar to what is told to happen in hypnosis: telling a person to do something after hypnosis, that person ( sometime) can do it after hypnosis without remembering the order.

Good Luck

There are a number of considerations to take into account when using your blocking reagent. Using whole serum looks attractive on paper as it contains several proteins, yet the major protein is serum albumen. If you use whole serum, there will also be IgG's in there, and it is for this reason that people use whole serum from the same species that was used to raise the secondary antisera [think what would happen if you were to use the whole serum from the same species in which your primary antisera were raised]. In the past I have used secondary pig antibodies, and normal pig serum was easily obtained from an abattoir. Most of my secondary antisera are made in goats and I use normal goat serum. For many antisera BSA will work just as well, however I never try that as I have many neuropeptide antisera in which BSA is used as the carrier protein and hence my primary antisera may contains anti-BSA IgGs. If anti-BSA IgG's were to become a problem other blocking reagents like the one suggested here (fish skin gelatin) or non-fat milk powder may work as well.

You can look up Mimetas and easily make a BBB model with their OrganoPlate https://mimetas.com/overview-mimetas-applications/human-blood-brain-barrier

There was a paper published in February this year in Scientific Reports with this platform: https://www.nature.com/articles/s41598-018-20876-2

Hope it helps!

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

Ok, after two years, I can come up with a partial answer!

References

1. Gossen A, Hahn A, Westphal L, Prinz S, Schultz RT, Gründer G, et al. Oxytocin plasma concentrations after single intranasal oxytocin administration - A study in healthy men. Neuropeptides [Internet]. Elsevier Ltd; 2012;46(5):211–5. Available from: http://dx.doi.org/10.1016/j.npep.2012.07.001

2. Dhakar MB, Stevenson EL, Caldwell HK. Oxytocin, vasopressin, and their interplay with gonadal steroids. In: Oxytocin, Vasopressin and Related Peptides in the Regulation of Behavior [Internet]. 2013. p. 3–26. Available from: http://ebooks.cambridge.org/ref/id/CBO9781139017855

3. Frayne J, Nicholson HD. Effect of oxytocin on testosterone production by isolated rat Leydig cells is mediated via a specific oxytocin receptor. Biol Reprod [Internet]. 1995;52(6):1268–73. Available from: http://www.ncbi.nlm.nih.gov/pubmed/7632835

4. Burnham TC. High-testosterone men reject low ultimatum game offers. Proc Biol Sci [Internet]. 2007;274(1623):2327–30. Available from: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1950304&tool=pmcentrez&rendertype=abstract

5. Zak PJ, Kurzban R, Ahmadi S, Swerdloff RS, Park J, Efremidze L, et al. Testosterone Administration Decreases Generosity in the Ultimatum Game. Aleman A, editor. PLoS One [Internet]. 2009 Dec 16;4(12):e8330. Available from: http://dx.plos.org/10.1371/journal.pone.0008330

6. Zilioli S, Ponzi D, Henry A, Maestripieri D. Testosterone, Cortisol and Empathy: Evidence for the Dual-Hormone Hypothesis. Adapt Hum Behav Physiol [Internet]. 2015;1(4):421–33. Available from: http://link.springer.com/10.1007/s40750-014-0017-x

7. Zak PJ, Borja K, Matzner WT, Kurzban R. The Neuroeconomics of Distrust: Sex Differences in Behavior and Physiology. Am Econ Rev [Internet]. 2005 Apr;95(2):360–3. Available from: http://www.jstor.org/stable/10.2307/4132847%5Cnpapers3://publication/uuid/BB68F281-21C7-452C-9238-C59CBE7560DA

8. Insel TR, Young LJ, Witt DM, Crews D. Gonadal steroids have paradoxical effects on brain oxytocin receptors. J Neuroendocrinol [Internet]. 1993;5(6):619–28. Available from: http://www.ncbi.nlm.nih.gov/pubmed/8680433

9. Arsenijevic Y, Tribollet E. Region-specific effect of testosterone on oxytocin receptor binding in the brain of the aged rat. Brain Res. 1998;785(1):167–70.

10. Bos PA, Hofman D, Hermans EJ, Montoya ER, Baron-Cohen S, van Honk J. Testosterone reduces functional connectivity during the ‘Reading the Mind in the Eyes’ Test. Psychoneuroendocrinology [Internet]. Elsevier Ltd; 2016;68(March):194–201. Available from: http://linkinghub.elsevier.com/retrieve/pii/S0306453016300671

11. Frith CD, Frith U. The Neural Basis of Mentalizing. Neuron [Internet]. 2006;50(4):531–4. Available from: http://linkinghub.elsevier.com/retrieve/pii/S0896627306003448

12. Goldstein JM, Jerram M, Poldrack R, Anagnoson R, Breiter HC, Makris N, et al. Sex differences in prefrontal cortical brain activity during fMRI of auditory verbal working memory. Neuropsychology [Internet]. 2005;19(4):509–19. Available from: https://www.tandfonline.com/doi/full/10.1080/10253890.2017.1378638

13. Güvendir E. Why are males inclined to use strong swear words more than females? An evolutionary explanation based on male intergroup aggressiveness. Lang Sci [Internet]. Elsevier Ltd; 2015;(13):1–7. Available from: http://linkinghub.elsevier.com/retrieve/pii/S0388000115000194

14. Mehta PH, Beer J. Neural Mechanisms of the Testosterone-Aggression Relation:The Role of Orbitofrontal Cortex. J Cogn Neurosci [Internet]. 2010;22(10):2357–68. Available from: http://www.ncbi.nlm.nih.gov/pubmed/19925198

Dear Yojet,

I suppose you have patch clamp rig to measure ionic currents, if yes, then you can simply patch NSC derived neuron and record inward I_Na and apply TTX. You shall see rapid and almost complete block of sodium currents (see attached pic) which will confirm the efficacy and potency of TTX stocks you have. You can refer to the following paper for internal external combination and protocol to be used for recording.

measuring

Ensure that you aliquote your TTX and store it at -20/80 to preserves its efficacy. I hope it helps!

Best,

Nand

Hi Lorena

Videos should be in .avi format. If you send me a mail I can share the toolbox with you.

Best

Julian

Did you ever try transfecting primary motor neurons with AMAXA?

Dear Zhou,

That will depend on the level of tissue and bleeding. If there is significant level of blood supply to the area that you are recording from, the reduction in local oxygen supply can cell death. Another factor is the swelling associated with damage to the Pia mater and damage other tissue ( interstitial) damage. Usually as you go down with your electrode you should be able to see/hear (from amplifier) discharges of damaged neurons. If your electrode tip is not blocked and you hear/see no discharges at all. That is bad news, meaning substantial damage. That is what I used to do in my studies, for an example see below.

Best wishes,

Refik

Just to avoid you a big waste of time; PC-12 Adh do not differentiate/grow neurites. I tried every possible experimental setups. They are not sensitive to NGF.

I dissolved the carbenoxolone disodium salt in distilled water. However, after mixed the carbenoxolone with saline solution, there is a lot of cloudy things. Does any one see this phynomena?

Hi Seungcheol Ahn,

Thanks for the update. That is a non-trivial slice, and one that I don't have personal experience with. My worry would be that you have cut the axons between the MNTB and the LSO.

If I were you, I would cut hippocampal slices. Pretty much any slice you cut that has a complete hippocampus in it will have a connection between CA3 and CA1. If you stimulate in stratum radiatum and put a recording electrode in the pyramidal cell layer you should see a large population spike (> 1mV). If you can not see this, then there is a problem with your stimulating or recording system.

However, if I had to make a guess, other than the worry that you have cut the MNTB LSO axons, it might be that the resistance of your stimulating electrode is too high. If we say that it has a resistance of 5MOhm, and you want to pass 100uA of stimulating current, then that would require 500 Volts, which is far more than any stimulating system that I know of has available. You'd be limited to less than 20 uA, which is starting to get pretty low (not that that amount can't work, it's just not what I would recommend for testing a system).

Try stimulating with a simple Tungsten wire (or something similar), with a resistance of 100kOhm or so.

Under the waveform tab you set up three waveform epochs, the first one is just light, the second one is light plus stimulus and the third one is just light again.

Vlad, probably the question is the sentence right after "I am wondering (...) whether...". Isn't it?   :v

 Hi Angelica,

Depending on the specifics of your experiment, you might also try doing transient transfection of a membrane bound fluorophore (e.g. mGFP https://www.addgene.org/14757/). I've done such transfections in on primary neural culture using CaPO4 method and Lipofectamine. In each case, the key was to optimize concentrations and incubation.

Best of luck, 

Joey 

Please check the material at the link I sent you on the previus message (this one: http://cienciaparaeducacao.org/eng/publications/)

Best,

Mateus

Most likely the homogenate contains glutamate as it is near millimolar concentrations in cytoplasm which will activate glutamatergic receptors in the pyramidal neurons. Glycine concentration is also likely sufficient to activate GluN1 subunits of heteromeric NMDA receptors. I suggest coapplication of ionotropic glutamate receptor antagonists, eg. CNQX and APV.

Hee-Hwan,

In my experience with brain derived neural stem cells, there are a number of things that affect the maintenance of the the stem cells and the development of the neurospheres. 

Cell passage: how high is the passage number of your cell line? The higher it is the less likely the cells will behave the way you want.

Expansion time: you say your are passaging the cells every 7 days or so.  How big are the neurospheres at this time?  the smaller the spheres, the better.  It is when the neurospheres get really big (1-2 millimeters in diameter) that the cells begin to differentiate.  When this happens and you dissociate the spheres, stem cells which have started differentiating down a glial cell fate will attach to the bottom of the plate.  The floating cells are either dead cells OR are still undifferentiated. When there are only a few of these floater cells, it might be better to re-plate them in a smaller dish, then expand them into a larger dish in 2-3 days (do not passage them, just dissociate when they are 10-15 cell spheres and put everything back into a larger dish).

Serum and growth factors:  Sometimes serum contains growth factors that encourage differentiation.  It is better to use a serum-free media and add growth factors that help maintain the undifferentiated state (ie. bFGF). 

It is good that you are not coating your dishes.  You might even try non-tissue culture dishes, low attachment dishes, or bacterial dishes to reduce attachment.

I've listed some good references below, although they are all brain based neural stem cells.  It is possible that spinal cord stem cells behave entirely differently.  What kind of cells are you hoping to get? glia? schwann? neurons? Gage's seminal paper in Science is a good place to start although keep in mind the field has advanced a lot since his original paper 17 years ago. 

Hope this helps!  Cheers, Kim

Gage FH. Mammalian neural stem cells. Science. 2000 Feb 25;287(5457):1433-8.

Review. PubMed PMID: 10688783

Rietze RL, Reynolds BA. Neural stem cell isolation and characterization.

Methods Enzymol. 2006;419:3-23. Review. PubMed PMID: 17141049.

Marchenko S, Flanagan L. Passaging human neural stem cells. J Vis Exp.

2007;(7):263. doi: 10.3791/263. Epub 2007 Aug 22. PubMed PMID: 18989434;

Kim HT, Kim IS, Lee IS, Lee JP, Snyder EY, Park KI. Human neurospheres derived

from the fetal central nervous system are regionally and temporally specified but

are not committed. Exp Neurol. 2006 May;199(1):222-35. Epub 2006 May 22. PubMed PMID: 16714017.

Dr. S. Liu,

Your answer was very much appreciated and I have learned a great deal due to your paper in PNAS.  I am going to be studying the db/db K strain model in the coming months, while I will be focusing my TEM  studies of this model primarily on the neurovascular unit I will also be evaluating the  dysmorphic myelin white matter and due to the better understancing of you PNAS paper I will be keeping a close observation regarding the assymettrical excitatory snapses as well.  You paper really helped and I am so thankful for your earlier response to my questions regarding an observation of increased excitatory synapses in the type 1 model and now will be alert to observe similar changes in the obese T2DM of obesity insulin resistance hyperinsulinemia  and persisitent elevation of glucose levels well above the diabetic range for mice  - approximately 300-400 ranges. 

Sincerely ,

M.R.(Pete) Hayden MD 

Dear Antant,

as a beginner in image analysis I found this book extremely helpful, practical and also entertaining:

I also have experience with matlab. Matlab can be connected with ImageJ and fiji:

So you can combine the two softwares. I like that, because I can immedately create the graphs as well. 

Good luck with your analysis!

OK.  I think it is a good idea.

Thank you for your timely response William! This does help a lot. Information about how well the cell can get signals to the the soma is still pertinent information in my study.