Selective astrocyte marker except GFAP?

More Christina Ahrensmeier's questions See All

Should I use PCR or Western blot?

I want to prove the existence of S100, S100A6, GFAP, MOG and MPZ in SG- and CN-sphere as well as in SG and CN tissue. What is more reliable; PCR or Western blot in this case? What do you think?

11 December 2015 3,901 4 View

error in running a model in Abaqus

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17 August 2021 0 0 View

error in modelling model in Abaqus

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17 August 2021 0 0 View

MGL tools

20 June 2021 0 0 View

i have some problem with chtMultiRegionSimpleFoam

--> FOAM FATAL ERROR: Maximum number of iterations exceeded From function Foam::scalar Foam::species::thermo::T(Foam::scalar, Foam::scalar, Foam::scalar, Foam::scalar (Foam::species::thermo::*)(Foam::scalar, Foam::scalar) const, Foam::scalar (Foam::species::thermo::*)(Foam::scalar, Foam::scalar) const, Foam::scalar (Foam::species::thermo::*)(Foam::scalar) const) const [with Thermo = Foam::hConstThermo >; Type = Foam::sensibleEnthalpy; Foam::scalar = double; Foam::species::thermo = Foam::species::thermo >, Foam::sensibleEnthalpy>] in file /home/ubuntu/OpenFOAM/OpenFOAM-4.1/src/thermophysicalModels/specie/lnInclude/thermoI.H at line 66. FOAM aborting #0 Foam::error::printStack(Foam::Ostream&) at ??:? #1 Foam::error::abort() at ??:? #2 Foam::heRhoThermo >, Foam::sensibleEnthalpy> > > >::calculate() at ??:? #3 Foam::heRhoThermo >, Foam::sensibleEnthalpy> > > >::correct() at ??:? #4 ? at ??:? #5 __libc_start_main in "/lib/x86_64-linux-gnu/libc.so.6" #6 ? at ??:?

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02 March 2021 0 0 View

Pete Williams

Christina:

A good option might be GLAST (sometimes called EAAT). This gets all astrocytes + radial astrocytes, without (to my knowledge) getting other glial cells. However, I don't know its specificity in the CN.

Christina Ahrensmeier

Hey Pete,

I looked it up. GLAST ist also expressed in schwann cell myelin, neurons and satellite cells of spiral ganglion and additionally oligodendrocytes.

Thanks for the hint.

http://citeseerx.ist.psu.edu/viewdoc/download;jsessionid=AC57526FE7CB231DEA3EB1F6A30A6687?doi=10.1.1.377.5713&rep=rep1&type=pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2667548/

Refik Kanjhan

Hi Christina,

What about vimentin. I know that it primarily labels radial glia and astrocytes. You may also consider targeting more than one marker. Perhaps using couple of markers, GFAP and vimentin together and GFAP and GLAST or EAAT2 together may help you. In the past I have found that TASK-1 and NHERF molecules also localized to the astrocytes, but I am noy sure how specific is that.

best wishes,

Refik

Hey Rifik,

thanks for your suggestions. I found that vimentin is expressed in the cytoskeletion of SC and neurons and TASK-1 is also expressed in SC but only in carotid bodies. So I will try to exclude astrocytes by a triple staining with S100, GFAP and MPZ. I hope that it may works.

best regards,

Christina

http://dev.biologists.org/content/139/7/1359.long

Article TASK-1, TASK-2, TASK-3 and TRAAK immunoreactivities in the r...

Sounds great and sounds like you know pretty much what you are. Cochlear nucleus is special to study with interesting its synaptic organization. Thank you for sharing it with us. I have previously used astrocyte labelling, but never had to do just astrocyte labelling. Looks like multiple labelling is currently one of your options. If I hear anythingelse I will let you know.

All the best,

Thank you, Rafik.

I will let you know, if the triple staining works.

Best regards,

Rogier Min

I have been using glutamine synthetase antibody which in cortex gives nice cytosolic astrocyte labelling. No neurons stained for sure. Not sure about other glial populations, but this paper suggests no strong oligodendrocyte labelling:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3797418/

Hi Roger,

I already looked GS up. It's highly expressed in differentiated SC.

But thanks for trying.

http://www.ncbi.nlm.nih.gov/pubmed/20107048

Marcin Siwiec

I've heard good things about Aldh1L1. It's supposed to be a highly specific astrocyte marker in the brain, albeit somewhat challenging to stain for.

Best wishes,

Marcin

Hey Marcin,

I'm sorry to say that Aldh1L1 is also a SC marker.

Thanks for trying.

http://jnci.oxfordjournals.org/content/106/8/dju184.full

Oh well, you got me :) Thanks for letting me know and good luck with the triple staining!

Ghulam Md Ashraf

really nice suggestions, thanks eveyone

Staining is done. It is not one of the best stainings but sufficient. I found single positive, double positive or triple positive cells. So there are Schwann cells (MPZ+/S100+/GFAP+; MPZ+/S100+; MPZ+/GFAP+) and also cells which are GFAP+/S100+. These cells are may be Astrocytes or Oligodendrocytes.

most probably astrocytes.

Chiara Ciriachi

NDRG2 is a new marker for astrocytic processes and cells (Flugge et al, 2014, Cell Tissue Res), but I have not worked with this again. Maybe you can try it.

Patro Nisha

You can try with Glutamine synthetase, an enzyme in astrocytes to convert glutamate to glutamine.

Hi Patro,

as I already mentioned GS is highly expressed in differentiation SC. But thanks for trying.

Serguei N Skatchkov

Dear Christina, indeed, we used several specific astrocyte markers (GS, GLAST, GLT-1, OCT-1, OCT-2, OCT-3, Kir4.1, Kir6.1,etc, however the staining is strongly dependent on age. For example, GLT-1, OCT-3, Kir4.1 and Kir6.1 are stably expressed in rat and mouse after 11-21 days, except if cells are transormed to cancer cells as glioblastoma.

If you are looking for healthy adults, try SLC22A-3 (organic cation transporter, OCT-3) or SLC18B1 (vesicular polyamine transporter, vPAt), or directly try spermine and spermidine. Check it. The markers would be very interesting for you. I can provide more information if need.

Cordially, Serguei.

Hey Serguei,

thanks for the suggestions. Some are promising. It would be nice if you could provide more informations.