Science topic
Neurochemistry - Science topic
The study of the composition, chemical structures, and chemical reactions of the NERVOUS SYSTEM or its components.
Questions related to Neurochemistry
Tell me the literature or articles where you can find the neurochemistry of forgiveness (which systems work at the same time)
I now that alot of artificial networks has appeared now. And may be soon we wil not read articles and do our scientific works and AI will help us. May be it is happening now? Wat is your experience working with AI and neural networks in science?
I analyzed the internal standard "2-methyl-L-cysteine hydrochloride" using HPLC and the chromatogram shows three peaks instead of one which is weird!!
I repeated the preparation process two times to check if there is a contamination problem but the chromatogram still showing three peaks. So what do you suggest?! Is there a possibility that the internal standard is converting to other compounds?!
I would like to scan calcium-indicator-dye-loaded neurons after fixation, just to know, which cells have been loaded. Fura and Carbodiimide fixation have been reported to retain fluorescence but I prefer paraformaldehyde fixation for several reasons. Is there a single calcium dye, which is not quenched after paraformaldehyde fixation?
The incredible thing about Physarum polycephalum is that whilst being completely devoid of any nervous system whatsoever (not possessing a single neuron) it exhibits intelligent behaviours. Does its ability to intelligently solve problems suggest it must also be conscious? If you think, yes, then please describe if-and-how its consciousness may differ {physically or qualitatively ... rather than quantitatively} from the consciousness of brained organisms (e.g., humans)? Does this intelligent behaviour (sans neurons) suggest that consciousness may be a universal fundamental related more to the physical transfer or flow of information rather than being (as supposed by most psychological researchers) an emergent property of processes in brain matter?
General background information:
"Physarum polycephalum has been shown to exhibit characteristics similar to those seen in single-celled creatures and eusocial insects. For example, a team of Japanese and Hungarian researchers have shown P. polycephalum can solve the Shortest path problem. When grown in a maze with oatmeal at two spots, P. polycephalum retracts from everywhere in the maze, except the shortest route connecting the two food sources.[3] When presented with more than two food sources, P. polycephalum apparently solves a more complicated transportation problem. With more than two sources, the amoeba also produces efficient networks.[4] In a 2010 paper, oatflakes were dispersed to represent Tokyo and 36 surrounding towns.[5][6] P. polycephalum created a network similar to the existing train system, and "with comparable efficiency, fault tolerance, and cost". Similar results have been shown based on road networks in the United Kingdom[7] and the Iberian peninsula (i.e., Spain and Portugal).[8] Some researchers claim that P. polycephalum is even able to solve the NP-hard Steiner minimum treeproblem.[9]
P. polycephalum can not only solve these computational problems, but also exhibits some form of memory. By repeatedly making the test environment of a specimen of P. polycephalum cold and dry for 60-minute intervals, Hokkaido University biophysicists discovered that the slime mould appears to anticipate the pattern by reacting to the conditions when they did not repeat the conditions for the next interval. Upon repeating the conditions, it would react to expect the 60-minute intervals, as well as testing with 30- and 90-minute intervals.[10][11]
P. polycephalum has also been shown to dynamically re-allocate to apparently maintain constant levels of different nutrients simultaneously.[12][13] In particular, specimen placed at the center of a Petri dish spatially re-allocated over combinations of food sources that each had different protein–carbohydrate ratios. After 60 hours, the slime mould area over each food source was measured. For each specimen, the results were consistent with the hypothesis that the amoeba would balance total protein and carbohydrate intake to reach particular levels that were invariant to the actual ratios presented to the slime mould.
As the slime mould does not have any nervous system that could explain these intelligent behaviours, there has been considerable interdisciplinary interest in understanding the rules that govern its behaviour [emphasis added]. Scientists are trying to model the slime mold using a number of simple, distributed rules. For example, P. polycephalum has been modeled as a set of differential equations inspired by electrical networks. This model can be shown to be able to compute shortest paths.[14] A very similar model can be shown to solve the Steiner tree problem.[9]"
source of quotation: https://en.wikipedia.org/wiki/Physarum_polycephalum
Background:
Professional-school student with extensive ACE (child adverse events) history along with severe depression and anxiety diagnosed over previous year, presented with recent severe ADHD (I-Type) diagnosis at age 26.
Documentation confirmed maximum dose step therapy for various Amphetamine-based stimulants was completed but still not found to be fully affective.
Unexpectedly, they are currently prescribed daily 50mg Mydayis (Mixed salts of single-entity amphetamine product) along with 80mg Prozac, and consumming 300-400mg of caffeine.
Due to initial medication-only use producing very minimal stabilizing effects, but found to increase at re-introduction of SSRI and further increase with Caffeine reintroduction.
No adverse effects (cardiac, neuromuscular, neurocognitive) have been reported/measured in 4 months of aforementioned therapeutic combination.
NOTE: Adverse reaction to methylphenidate-based medications were identified early on.
Assessment of (remaining) presenting symptoms seems to overlap with tentatively defined SCT Criteria.
NOTE: Student has never been prescribed Strattera (only presently confirmed SCT-symptom relief medication)
Specific question:
Recent research has shown SCT + ADHD to correlate with much greater impairment in adults, do you think a combination of severe ADHD + SCT may result in required use of excess pharmacotherapy dosages that surpass established safe therapeutic/combination parameters?
Hi everyone ^_^
I was wondring if there is a specific technique to study the chemical change responsible for DID and I want to know if I could follow a specific procedure to the neurochemicals behinde this disorder and how the alteration vary between a normal person and a DID patient? also, the most important thing is finding a specific drug to cure this disorder. I know that there are drugs that help the patients endure anixty and depression come with this disorder but there is no direct and specific cure for DID.
I read that contextual treatment helped the patient somehow to know themselves and some of them return the control and can change from one personlaity to the other when they want but it didn't work for most of them although it benfits them somehow but not cure them. So; I read a lot of artiles and reviews from the litreture but most of them focused in the contextual treatment not chemical based treatment!!
I don't know if this is right or not but I think this return to neuropsychopharmacology and I'm a chemistry master student so I want to know if there is a possibility to study this in my thesis project and if I need a high tech equipments to do that?
The Elevated Plus Maze Model (EPM) is basically tested for the anti-anxiety activity. Currently, I am working on learning and memory activity in rats by the Barnes Maze and EPM test. Is the EPM reliable for nootropic activity? whether the electrophysiology of hippocampus part of rats brain suitable or not to know the mechanism of action for tested drugs?
Does anyone know how/if its possible to measure erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) in living humans?
Currently doing a research project for my Honor's Chemistry class, lately I've been looking into Brain Machine Interfaces on the side for research later on this summer, however, I was hoping to find a relationship between BMI's and chemistry so that I can use this project as an addendum for my internship.
The scope of the project is essentially on current research in chemistry, or historical importance of chemistry.
that being said I'm aware there is some chemistry involved in BMI's however, I can't find anything to specifically research on.
Any ideas or advice?
GABA can be released via GABA Transporter (GAT) through reversion of this transporter. But how this 'reversion' works? In which situation this transporter starts to release (and not uptake) GABA? Is the mechanism described?
Can systems dynamics (e.g. Braun, 2001) and Fisher’s account of temperament (Fisher et al., 2015; Brown et al., 2013), be used to explore the "emergence" of values? For example, a child developing in a setting which rewards and reinforces curiosity and creativity might become increasingly sensitised to feelings associated with their dopamine system (Fisher, below), and subsequently seek experiences which offer that particular feeling of reward. This might establish a reinforcing cycle (Braun, 2001), through which the child’s temperament becomes predominantly expressive of the dopamine system. From Schwartz’ human values perspective would this child’s goal choices tend to consistently express openness to change, independence and self-direction values? Would the child’s choices be values-based insofar as they are infused with feelings, refer to desired goals, and exhibit consistency that transcends specific situations (Schwartz, 2012)?
Might Fisher’s Explorers express Schwartz’ Openness To Change values, Builders express Conservation values, Directors express Self-enhancement values, and Negotiators express Self-transcendence values?
Might accounts connecting neurochemical systems with values and feelings (e.g. Lövheim, 2012) improve the utility of the worthwhile, satisfied, happy, anxious and social trust dimensions in the subjective wellbeing literature (e.g. Michaelson et al., 2012) and enrich the goal choice (e.g. Knafo and Sagiv, 2004) and productivity literatures (e.g. Parks and Guay, 2009)?
Fisher Four broad temperament dimensions:
· Explorers expressive of the dopamine system linked with traits including: being curious, creative, spontaneous, energetic, risk-taking, novelty-seeking, mentally flexible.
· Builders expressive of the serotonin system linked with traits including being: traditional, conventional, following the rules, respecting authority.
· Directors expressive of the testosterone system linked with traits including being: analytical, logical, direct, decisive.
· Negotiators expressive of the oestrogen system linked with traits including: being empathetic, emotionally expressive, good with people.
Schwartz (2012) values super groups:
· Openness to change values: emphasise independence of thought, action, and feelings and readiness for change (self-direction, stimulation)
· Conservation values: emphasise order, self-restriction, preservation of the past, and resistance to change (security, conformity, tradition)
· Self-enhancement: emphasise pursuit of one's own interests and relative success and dominance over others (power, achievement).
· Self-transcendence: emphasise concern for the welfare and interests of others (universalism, benevolence)
References
Braun, W., 2001. The Systems Modelling Workbook. Berlin: Springer, 2001. [Online]. [Accessed 19 January 2017]. Available at http://www.albany.edu/faculty/gpr/PAD724/724WebArticles/sys_archetypes.pdf
Brown, L.L., Acevedo, B. and Fisher, H.E., 2013. Neural correlates of four broad temperament dimensions: testing predictions for a novel construct of personality. PloS one, 8(11), p.e78734.
Fisher, H.E., Island, H.D., Rich, J., Marchalik, D. and Brown, L.L., 2015. Four broad temperament dimensions: description, convergent validation correlations, and comparison with the Big Five. Frontiers in psychology, 6.
Lövheim, H., 2012. A new three-dimensional model for emotions and monoamine neurotransmitters. Medical hypotheses, 78(2), pp.341-348.
Knafo, A. and Sagiv, L. 2004. Values and work environment: Mapping 32 occupations, European Journal Of Psychology Of Education, 19 (3).
Michaelson J., Mahony, S. and Schifferes, J. 2012 Measuring Well-being: A guide for practitioners, London: New Economics Foundation.
Parks, L.,and Guay R. 2009. Personality, values, and motivation, Personality and Individual Differences, 47, pp. 675–684
Schwartz, S. H. (2012). An Overview of the Schwartz Theory of Basic Values. Online Readings in Psychology and Culture, 2(1). http://dx.doi.org/10.9707/2307-0919.1116
Hi everyone,
I have an Amyloid beta 1-42 fragman, but i dont know how can I prepare this. In water or another liquid. There are not enough paper in web about that. I asked to many people, but they did not give an answer clearly.
I want to inject AB 1- 42 into the brain parenchyma.
Is there any advice?
What is the normal range NT-proBNP using ELISA kit in the healthy adolescents?
Reference. please
Dear all,
I am currently working with ghrelin (a gut peptide) and I would like to check if the peptide crosses the blood brain barrier and if it is possible to find out in which areas it reaches. I know that the question is a bit general, but I've been puzzled, since there are no previous studies investigating that question and there are only some hypotheses out there which don't seem to help me. Ideally, I would like to somehow trace "the flow" of the peptide through the BBB and its distribution across the brain areas.
Any ideas or suggestions would be highly appreciated!
Thank you all in advance.
Regards,
Lydia
It has been known for a long time now that cotinine, a metabolite of nicotine, is a nootropic and more importantly, is somewhat neuroprotective for PD and AD. It is also known that cotinine is well tolerated by human subjects and has none of the negative side effects of nicotine itself. Given today's great concern regarding both PD and AD, how can it be that research on this compound, very closely approximating a "silver bullet" if you will, is not being conducted with great intensity by NIH and/or the pharma industry? Why does this seemingly magical solution to so many problems sit on the side while less practicable molecules get to dance?
I don't get it. There is tons of research supporting this but not much is happening. Why?
Brain tissue was homogenised in 25 mM Tris - 4 M urea buffer. After a freeze thaw cycle noticed that the tissue formed clumps. Which is not getting dissolved. Why it is so?
Just recently, features of psychiatric disorders such as schizophrenia were suggested to be due to a decrease of brain memory center’s inhibitory neurons. HSV-infection is primarily limited to mucosal epithelial cells and neurons, and autophagy is critical in antiviral defense in neurons. Given that type I IFN treatment failed to completely block HSV-1 replication and induce cell death in neurons, some xenobiotical factors might be considered triggering replication of neurotropic HSV-1, and lead to a loss of inhibitory neurons.
As we know, microdialysis is an acceptable method for detecting the concentration of many metabolites, such as lactate, glucose, glutamate etc. in human brains. However, I wonder weather this technique shrank the true value for which the taken interstitial fluid was diluted in this process. Thus, could you provide some evidences to clarify the true value of C. lactate in human brain?
Orch-OR theory for consciousness asserts that the microtubules are the neural structures that support the quantum effects. Let's assume that it is true. Therefore, if they have to play a role in the brain, they need to effect the signal transmission in the brain. Is there any indication for such an effect?
I used neutrophil elastase antibody to stain brain sections and got a lot of background staining. I used 3% H2O2 and 2% serum blocking.
Hello!
I am looking to utilize ifenprodil in slices of rat cortex. I have read that it can also act on α-adrenergic receptors which I wish to avoid. Most work seems to utilize 0.1-10 micromolar, and I see EC50s of approx. 0.3 micromolar. Does anyone have experience with using ifenprodil in slices and have a recommendation of a selective concentration?
I have had this ICC working for several years and have used it for multiple experiments, but recently it has stopped working. I am now having nearly every cell in the brain labeled in every trial I do. I initially tried ordering a new lot of the primary antibody, but that made no difference. I have modified the concentration of the secondary and the time in the DAB and neither of those helped. I tried doing a no primary control to rule out nonspecific binding of the secondary and a no primary or secondary control to rule out ABC binding to endogenous biotin in the tissue. Both of those trials came out clean, with no staining, suggesting that the problem is not with the secondary or the ABC. We have tried replacing the paraformaldehyde (our brains are not perfused, so fixation is the first step of our IHC), the methanol, the H2O2, the normal goat serum, and the DAB tablets and none of these things have solved the problem. We also thought that contamination in the water may have been the problem, but I got the same result when using all solutions made with HPLC grade bottled water. Finally we thought there may have been a problem with the tissue since it had been in the -80 freezer for 2+ years and may have experienced temperature variation, but I recently included sections from a brain that was collected and sliced within the previous week and that did not solve the problem. I have run out of ideas for what could be causing the excess staining. Does anyone have suggestions on what I could try?
I need to know where the cholinergic afferents that reach the temporal side of the perirhinal cortex are coming from. Thanks.
Dear expert,
I need some works about diffusion mechanism (How molecules are transferred from blood into the brain tissue) in the brain .or an up-to-date brain anatomy.
Since, the primordial Na-pump on the plasma membrane is the ultimate source of energy of animal cells; it appears that the remaining Ca-signaling networks will join the leading Na-pump, and the potential energy is used by remaining networks for homeostasis. Hence, the cellular bioenergetics initiated and managed by the NaAF regulated Na-pump, by turn regulated by the provisional Ca-pump and Ca-signaling, is the leading-center of a cell’s holistic-networks.
We visualized operation of the ion-pump under Ca-signaling as an all-time allosteric dancing of the Na-pump. During operation, the ubiquitous NaAF acts as the dancing-partner cum gate-keeper of the double-gated (two α-subunits) Na-pump allowing simultaneous transport of Na (in) and K (out), while the process is controlled by Ca-signaling. Intensity of the dancing will be affected by the isoform-nature of the Na, K-ATPase which varies in various bodily cells in a tissue-specific manner except brain where the Na-pump is area-specific. The extremely specialized functions of the area-specific brain (of human) are of prime interests now-a-days.
The Ca-signaling is conducted periodically by the provisional Ca-pump which is an altered form of the cell-specific Na-pump. The provisional Ca-pump is entrusted to pump out the excess (inhibitory) local Ca for continuation of Na-pump function for homeostasis, thus making it a stop-and-go type dance.
This brings us to the question we are asking. Are the molecular natures of the provisional Ca-pumps the same as the isoforms of the PMCA well-known in the vast Ca-ATPase literature?
It will be very interesting issue to investigate.
Can I know if I can detect calcineurin levels in brain of hamsters after prion infection in fresh sacrificed animal brain only or is it possible to detect levels after the freezing of brain sample for future analysis? I would like to use calcineurin activity assay kit.
Hello, I am trying to find a source of Deoxynivalenol (vomitoxin) HRP conjugate and having no luck. I could make it, but am hoping to find a commercial supplier. Anyone ever come across this?
The concentration of alpha synuclein is 25-100 micromolar and i use 800 rpm, 37 degree celsius. I am concerned about the sample volume (30 microlitres in 1.5 ml tubes) and whether agitation helps at small volumes.
I'm successful in separating the presynaptic and PSD fraction using a method that published in a Neuron article. I have some native gel running complication with the pre-synaptic membrane fraction as the gel goes bizarre. I found that the triton X-100 in the presynaptic fraction causing this while running native page gel. I even found the same while running in SDS/PAGE but I was successful after subjecting the presynaptic fraction to acetone precipitation. But the acetone precipitated presynaptic fractions is not compatible to resolve you native proteome as acetone disturbs the lipid bilayer thus no longer the native condition of your membrane proteins maintained. At the separation step of presynaptic and PSD fraction we use 1% triton at pH 8. During this step the presynaptic fraction is resolved in the supernatant along with 1% triton. I used to filter (used 100 K esp for running native page also used 10 K to 30 K filters) and exchange the presynaptic fraction buffer composition (ph 8 to 7.4) to get rid of triton but instead it gets concentrated to form a micelle I persume so. How do I get void this triton from this fraction without affecting the nativity of a proteome? Does anyone have any idea how do I over this situation?
I’m working with a new Haas type interface chamber, called BSC-HT Brain Slice Chamber System Haas Top (Harvard Apparatus). The problem I encountered is as follows: after calibrating the perfusion, the slice doesn’t look dry, yet the fEPSP amplitudes facilitate during the recording (increases by about 400 uV in an hour).
The ACSF is saturated with carbogen and is at room temperature, carbogen is also directly flown into the chamber to produce carbogen-steam. I tried different perfusion speeds (1-4 ml/min), but it didn’t make significant difference. I previously worked with a similar Haas type chamber, and managed to overcome similar difficulties.
I wonder what may couse the facilitation in the fEPSP amplitude - any suggestion is welcome.
Thanks in advance.
I've been using trizol to homogenize frozen nerves. I've pooled 2, 4, and 6 nerves and ran qPCR for GAPDH. The Ct values for all combinations were about the same (~23). So I used 2 nerves to run a data set, but this time my Ct values for GAPDH ranged from 26 to 31. I think I may just not have any RNA, and the readout of the RNA concentrations is something else. What protocols have worked for others?
I have been staining brain sections of humanized mice with a number of human microglial and macrophage markers but nothing is showing up. The same antibodies are positive for peripheral tissues. Has anyone looked at human macrophage markers in the brains of these mice. These mice are supposed to have human CD34+ CD45+ and CD68+ cells in the brain. But I have failed to see any of these yet.
Hi all,
I am interested in AMPAR mediated mini EPSCs in hippocampal neurons. My current recording configuration (hibernate E as bath solution and Cs gluconate as internal) seem to allow me to record from them up to day in vitro (DIV) 25 for about 30 min with fairly stable access. The sad story is that I do not see a lot of mini events (1 - 2 events every 3 - 5 seconds --> much less than 1 Hz). I can see quite a lot spontaneous events starting at DIV 11 already (1 - 2 event every second or so). Does it sound like something you experience before? How would you recommend troubleshooting it? Maybe, like, changing the recording condition or culture condition to have more mini AMPAR EPSCs? Thanks a lot!!!
Best,
Huong
Below are some more information if you would like to know....
When the neurons are younger (Div 12 - 15), there are a lot of action potential driven EPSCs [huge events, > 100 pA]. And when they get to Div 25, there are mostly very small events (20 pA, more or less). The small events decay time is approximately from 4 - 30 ms.
Regarding the culture, Coverslips are coated with 1 mg/ml Poly D lysine. I plate the neurons from E17 - E 18 hippocampi at 1.4 millions neurons per 100 mm dish containing 6 coverslips. The coverslips are submerged in serum containing media. The coverslips have wax feet so I can flip them up side down into 60 mm dishes with neurobasal + B27 + glutamax + 20 % media conditioned by astrocytes [which facilitates the growth of a lot of astrocytes underneath the neurons].
Sometimes I also co-culture the neuron coverslips in dishes with astrocyte feeder layers [in which the cells are not touching each other i.e. Banker culture]. The viability and development look fairly good. I can see a fairly dense network of dendrites already at DIv 11 and it just gets denser over time. Cells are evenly distributing across the coverslip, not much fasciculation or any sign of substrate problem. I feed them twice a week after the first week.
In the past we have used Diasorin's ACTH IRMA for assays in human and rhesus plasma. This kit has been discontinued, and I am looking for a replacement. I have tried MP Biomedicals RIA and MD Biosciences ELISA. The ELISA has given more comparable results, but many of our samples are near the limit of detection which results in high variability in duplicate measurements. Does anyone have any other suggestions for a replacement plasma ACTH assay?
I am need to estimate the level of serotonin in my sample, for the that I have to compare my results with standard curve. I had tried lot of methodology but failed. So please suggest the working methodology (for serotonin standard by using serotonin creatinine sulfate monohydrate) to come up with good results.
Thanking you in advance for your kind courtesy.
I am performing patch-clamp in whole brains in vivo and including a fixable intracellular dye
In order to locate the cell I have recorded from more easily, I figured if I add some Hoechst to the intracellular solution, then; as it is highly fluorescent, membrane-permeable and does not interfere with living cellular functions, it would serve as a general locational marker.
Has anyone else tried this? I am expecting to see a nice UV patch in the cortex I have recorded from that should guide me to where the cell I have patched is :D
I'm patch-clamping horizontal cells from goldfish retinas, and the membrane potential is constantly reading 10-30 mV (from "I = 0" setting on my AxoPatch 200B amplifier). I don't know if it's physiological, pathological, or a problem with the equipment. When I put on glutamate, the cells repolarize towards the glutamate reversal potential (0 mV), so I think it's not an equipment problem - but I'm all ears if you have advice/ideas.
I've tried less papain during dissociation; this didn't change anything. I've tried whole cell and perforated patch over and over - no difference. I've replaced solutions and tried different ones. I've checked my osmolarity and tried different ones. I've tried new batches of fish. Any ideas?
Please help! You'll be my hero!
I'm interested in recording fiber volleys in CA1 elicited via stimulation of Schaffer collaterals in Organotypic slices. I have been looking around for an explanation of what type of stimulation electrode to use and how to place the stimulator/recording electrodes but have not been able to find a clear explanation.
Should I pull large glass pipettes for the recording electrode so that I have a lower resistance? Or does this not matter? Should I place the concentric bipolar electrode directly into the slice or should I place it in a glass pipette?
Thanks for your help.
I'd like to perform an experiment on mice inducing EAE with whole myelin as the immunogen. The most important reference seems to be "Oct;21(4):749-57.
Myelination in rat brain: method of myelin isolation. Norton WT, Poduslo SE.J. of Neurochemistry 1973" that obviously is about rats.. Thank you in advance for your help.
I am planing some experiments to study difference in basal synaptic transmission accross different animal conditions. I am thinking about producing a fiber volley vs fEPSP slope curve, but I haven't done this before.
Can anyone advise me for this exercise?
Regarding the x-axis of this putative curve, should I give fixed increasing steps for the stimulation intensity (for me this option seems easier) or should I try to find representative FV values (e.g. 0.05, 0.1, 0.15, 0.2, etc...; for me this option seems harder, since the FV size changes among preparations).
It would be great if you can help me.
Best regards,
Diego Fernández
Hi, nowadays i am trying to apply channel antibody into cerebellar Purkinje cell. However, every time I tried, whole cell condition went bad in 10 minutes (even initial condition was quite good).
Cerebellar Purkinje cell, especially its dendrites, is rather larger than other cells, so I should wait at least 20 minutes until the internal solution spread out enough.
I can see the common problems in Rs configuration; decreasing holding current, increasing leak current by time.
If you know any know-how, please let me know
thank you
The values of the ionic conductances for excitatory neurotransmitters can be found from the paper published by A. L. Hodgkin and A.F. Huxley. Are the values same for inhibitory neurotransmitters? Is yes, then what is the difference?
Since neuropeptides are small proteins and the samples we collect are very small. I tried some commercial kits that can separately get RNA, DNA and protein from the same sample. But I polled a lot of these samples together and then measured the concentration of whole protein. Even though the concentration of protein can hit the minimum amount used for western blot, no neuropeptides can be detected. I guess most of the neuropeptides either had already filtered through during extracting them by using the kit or the concentration were too low to detect. Is there any good way to handle this? The punch that I use is 0.75 mm in diameter and 0.3 mm in depth. Thanks.
I am planning to determine acetylcholine and glutamate level in brain tissue homogenate. Firstly I want to know whether it is possible to detect nerotransmittres in brain homogenate? Secondly if it's possible then how can I get rid of the fats and other proteins in brain homogenate to prepare a sample, that contains only nerotransmitter, to be run in HPLC. As all the papers, I found, to detect neurotransmitters in brain use brain dialysate.
I have no experience whatsoever transfecting cells with cDNA, and I am looking for help/guidance/knowledge from the community.
What electroporation unit do you recommend? I will be using mainly PC12 cells.
Are there particular settings or solutions you like?
Does anyone have any clue why the frequency of mEPSCs is so low (<0.1Hz) in rat cultured cortical pyramidal neurons at 14 DIV, even though sEPSCs activity is robust?
We usually generate cortical neuronal culture from E18 embryos and plate the neurons with a density of 10^6 on PDL/Laminin coated coverslip. The neurons were mainteined in BME supplemented with Pen/Strep, Glutamax, FBS, N2 and 2-Me. The medium was replaced every 2-3 days. We transfected the neurons with lipofectamine 2000 at 8DIV and started to record from 14DIV.
For the transfected pyramidal neurons (GFP positive), they have normal morphology including beautiful spines, and robust spontaneous EPSCs (~1Hz). However, mEPSCs frequency is always low. In some neurons, we can't see even one event in 100 second recording. We also tried to culture the neurons up to 21DIV, but the mEPSCs frequency didn't increase over the additional week.
I am trying to dilute DPCPX (A1 receptor antagonist) to inject in rats. According to some publications it can be done in a 10% DMSO, 90% saline solution. I tried this and was able to dilute the drug in DMSO, but whenever I add the saline the drug precipitates.
It has been said that some neurons have the capability to spontaneously generate an action potential without receiving any kind of synaptic input, in other words the AP is intrinsically generated.
How does the cell do this? Is this a slow depolarization resulting in tonic low frequency firing? Can phasic firing also be intrinsically generated?
Which brain areas and cell types are particularly known for exhibiting this type of behavior?
What is the best/optimal way to freeze the rat brain - liquid nitrogen, CO2, isoamyl alcohol? When you take out brain from 30% sucrose, what is the next step?
Can anyone tell me about what are the ethical issues in Human Neuroscience studies. Which are Drugs we cannot use? Which are parameters we can use to study Human Brain?
We tried Santa Cruz but there's a lot of non/specific staining. How about for PSD-95 Western blots?
I want to do a western blot using proteins from rat cerebral cortex but I am asking if there will be enough cells that express this?
I am working on a methylating protein and would like see the expression of gene in mature oligodendrocytes. What is the best method for transient but long lasting expression?
I am using a HCN-2 cell line, the growth rate of this cell line is greater than 120 hours. According to ATCC the growth rate of HCN-2 cells is stimulated by treatment with phorbol esters, but I am not getting a good growth rate after treatment with phorbol esters.
I'm working on the regeneration of the auditory cortex in adult rats. In my material, a restricted lesion induces axonal growth from the contralateral cortex, shown using injection of neuroanatomical tracers. Can any suggest antibodies for labeling axonal growth cones in histological sections?
What are the important points to be noted while looking at the uncoupled anion currents? The assay will require transfection and over-expression of a membrane protein. A ligand induced current will be looked at. Any inputs regarding these kind of assays?