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
What species?
In rodents I would synthesis a 14C labelled Ghrelin peptide, then after injection I would sacrifice the animals, section the brains on a cryotome and expose sections to photographic film to see where the peptide ends up.
in man it would be quite difficult, but essentially you would have to spin-label it for subsequent MRI scanning.
As Mostafa said earlier, you might just want to inject a large quantity (even unlabelled), then extract the CSF and see if the concentration of Ghrelin has increased from baseline.
You could fluorescently label the peptide with a small fluorophore, such as FITC. There are a number of commercially available kits which you could buy to do this. Then administer the peptide or perform in situ brain perfusion and fix the brain with paraformaldehyde. You can then look at its distribution using fluorescent microscopy. You can check whether the peptide reaches in an in tact form or not by running a brain homogenate from an unfixed brain on a gel. If you see the labeled peptide at the correct molecular weight this would indicate it reached the brain in tact.
Hi everyone,
Thank you for your time and your replies. Unfortunately the "Human Digestive System" was tagged automatically, my question was referring to rodents.
Your ideas have been very helpful and I will look into the available techniques more thoroughly.
Good luck to all!
Best,
Lydia
Hi,
I think that the best way to reach Prof. William Banks ([email protected]) who works on peptides and BBB for several decades and he has been investigating behavior of peptides, and how they across to brain.
Good luck
Mehmet
Hi Lydia,
Interesting topic. Please see Diano et al.2006 (Nature Neuroscience 9, 3, 381-388) where radio-labelled ghrelin was used to determine entry of peripheral ghrelin into the CNS. They also compare levels across different brain regions. I hope this helps.
Good luck.
Jeff
go through this paper
Brain circuits mediating the orexigenic action of peripheral ghrelin: narrow gates for a vast kingdom
Agustina Cabral, Pablo N. De Francesco and Mario Perello
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