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Just to keep topics separate, Brian.
I would be grateful for comments on the staining I achieved on FFPW section after HIER on adult rat brain, hippocampus. The nuclear staining is localised to just a few areas. Mainly hippo but clearly also, what I think is the Pars nervosa. At 1/100 I also see nerve fibre staining....is this non-specific? I have also immunostained rat/mouse E17 body T/S at cerv/thor/and lumbar regions and see the occassional positive nucleus.
Antibody from RELIATech #102-PA30S
Pics in gallery ( also seeing if I can get the hang of putting one here)
Thanks
carl
Thu Apr 27, 2006 10:26 am
brain
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I also like to add more information about Prox-1 marker for hippocampus.
I knew clearly when this marker used in hippocampus, it stains well with the progenitor cells of granule cells in the Dentate gyrus of adult mouse.
Thu Apr 27, 2006 3:59 pm
Carl
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Post subject: Thanks for info, Brain
OK....looking at my pic....do you think that all the nuclei that are positive are progenitor cells? It seems to me that the nuclei that are positive are granule cells..which are not progenitor cells? If I immunostain for DCX a very much smaller subpopulation of cells WITHIN the granule cells are positive...not the granule cells, per se
Would you agree that the progenitor cells in the sub-ventricular region of the anterior/lateral ventricle ( that migrate to form/replenish the olfactory neurones in mouse/rat)should also be positive?
Be most grateful for your comments.
Many thanks
Carl
Thu Apr 27, 2006 7:13 pm
brain
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I don't find your pic yet? Can you show me where they are?
But I'm pretty sure that the Prox-1 stain with the immature granule cell in hypocampal dentate gyrus. You can reference at Brandt et al at , Mol&Cell Neurosci 24 (2003) 603-613, or a comparative review
"Laminating the hippocampus", Eckart Förster, Shanting Zhao and Michael Frotscher, Nature Reviews Neuroscience,8 March 2006
I also agree with you that DCX is positive with a smaller population of cells within the granule cells. Recent review show that the DCX is mostly positive with progenitor cells, and Prox-1 is positive with progenitor cells but also with a mature granule cell as well. To prove this, some paper used BrdU staining that show the overlap staining of DCX with BrdU as well as a part of Prox-1 cell in adult hippocampal dentate gyrus.
Thus, The DCX is also overlap with Prox-1. This might suggest the DCX as a marker define from stem cell --> progenitor cells, the Prox-1 as a marker from progenitor cell-->mature granule cells.
As for the progenitor cells in the SVZ pathway to the OB, I am pretty agree with this opinion because I saw it already, but I not sure the differntiation of progenitor cells to form OB granule cell, mitral or periglomerular cells in OB in adult mouse . I have never seen the prove!
If you know it, show me!
Fri Apr 28, 2006 2:34 am
brain
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Dear Carl,
I found some pic in image gallery, i guess that yours?
As I though rightly, that the DCX positive under the granule cell layer where of the progenitor cell of gc (Pic: DoubleCortin in rat Hippocampus after P'Shop enhancement) and Prox-1 staining mostly granule cell layer, where of the mature gc. However, the Prox-1 might also overlap the DCX as well. Do you think so?
Fri Apr 28, 2006 2:45 am
lulbom
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Hi Carl,
The Prox1 staining in the hippocampus looks perfectly fine to me...and as many others have pointed out, Prox1 labels the progenitor cells as well as the post mitotic granule cells (both immature and mature).
I am not really sure abt Prox1 in other parts of the brain...so I cannot comment on that!
Mon May 01, 2006 7:28 am
Carl
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Post subject: Thanks for responses, Lulbom and Brain
Is Prox1 positivity only found in the nucleus?
I have to admit that I do not see the same cells expressing both the proteins, so Brain, perhpas you are right. Interestingly, I do not see ANY Nestin-positive cells in the rat Hippocampus...only DCX-positivity. Any idea why not?However, in the SVR of the anterior ventricle one sees both DCX and Nestin positivity ( in the same cells or not?).
Also, none of the hippocampal cells are positive for PSA-NCAM ( I have stained sections with two different PSA-NCAM Abs; the SVR prog. cells, however, are clearly positive)
Early days for me as my main area in one project is to define DAG lipase/CB1/2 expression patterns in the SVR
I will have to do serial section DAB staining but, better, double- IF staining next week to find out whether there is co-expression of DCX/PROX1 in the Hippocampus
NB: Brain- regarding the fate of the SVR progenitor neurones, I thought that they migrated to the olfactory epithelium and did not replenish the OB?
Mon May 01, 2006 1:47 pm
brain
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Hi Carl,
Which ages of mouse you do DCX, and Prox-1 or Nestin?
I do not read any papers demonstrate they replace the OB neurons, but I think it might occur.
I think you can try the BrdU with glial and neuronal marker to prove it.
Wed May 03, 2006 2:00 am
skku
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Post subject:
hhehe I would like to ask a small question,
Do the Prox-1 and Dcx stain with migrating neuron in the dgm1 and dgm2 (dentate gyrus migration) in which populate granule cell layer during late embryonic to postnatal day in mice? _________________ Seeing is believing!
Thu Dec 07, 2006 2:10 am
Carl
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Post subject: Hehe.....skku
Is that a rhetorical question? Or do you know the answer? I do not have an answer as I have not tried this Prox1 on developing mouse brain hippocampus.
The fact that most of the nuclei are positive in hippocampus of adult mouse pwax sections worries me , as Prox1 is an accepted marker for endothelial cells which are committing to become lymphatic vessel lining cells?Am I wrong? Perhaps my results are non-specific?
I still haven't found any info regarding Prox1 positivity in CNS.
Thnx for your comment
Best wishes
carl
Nb: I have posted pics in the IHC Image gallery
Thu Dec 07, 2006 8:14 pm
Carl
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Post subject: Explanation:
Sorry, I know that Prox1 is found in areas of the CNS...I just have not found any reference to it's presence in the nuclei of the dentate gyrus cells, to the extent that is shown in my pic, in the gallery. From that pic, I would assume that these are mature cells: why are they expressing Prox1, is what I meant
Carl
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. I have never seen the prove!
