Neuropeptide delivery to the brain: a von Willebrand factor signal peptide to direct neuropeptide secretion.
BACKGROUND
Multiple neuropeptides, sometimes with opposing functions, can be produced from one precursor gene. To study the roles of the different neuropeptides encoded by one large precursor we developed a method to overexpress minigenes and establish local secretion.


RESULTS
We fused the signal peptide from the Von Willebrand Factor (VWF) to a furin site followed by a processed form of the Agouti related protein (AgRP), AgRP(83-132) or alpha-melanocyte stimulating hormone. In vitro, these minigenes were secreted and biologically active. Additionally, the proteins of the minigenes were not transported into projections of primary neurons, thereby ensuring local release. In vivo administration of VWF-AgRP(83-132), using an adeno-associated viral vector as a delivery vehicle, into the paraventricular hypothalamus increased body weight and food intake of these rats compared to rats which received a control vector.


CONCLUSIONS
This study demonstrated that removal of the N-terminal part of full length AgRP and addition of a VWF signal peptide is a successful strategy to deliver neuropeptide minigenes to the brain and establish local neuropeptide secretion.
de Backer
Marijke W A
MW
Rudolf Magnus Institute of Neuroscience, Department of Neuroscience and Pharmacology, Utrecht University Medical Centre Utrecht, Utrecht, the Netherlands.
Brans
Maike A D
MA
Luijendijk
Mieneke C M
MC
Garner
Keith M
KM
van den Heuvel
Dianne M A
DM
Pasterkamp
R Jeroen
RJ
Adan
Roger A H
RA
D054369:Agouti-Related Protein / Q000096:biosynthesis / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D000229:Dependovirus / Q000235:genetics; D018014:Gene Transfer Techniques; D057809:HEK293 Cells; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007763:Lac Operon; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics / Q000502:physiology*; D044105:Receptor, Melanocortin, Type 4 / Q000096:biosynthesis / Q000235:genetics; D000521:alpha-MSH / Q000096:biosynthesis / Q000235:genetics; D014841:von Willebrand Factor / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
D054369:Agouti-Related Protein; D018076:DNA, Complementary; C470072:MC4R protein, human; D009479:Neuropeptides; D021382:Protein Sorting Signals; D044105:Receptor, Melanocortin, Type 4; D014841:von Willebrand Factor; D000521:alpha-MSH