TY - JOUR
T1 - The Prion Protein Regulates Synaptic Transmission by Controlling the Expression of Proteins Key to Synaptic Vesicle Recycling and Exocytosis
AU - Peggion, Caterina
AU - Stella, Roberto
AU - Chemello, Francesco
AU - Massimino, Maria Lina
AU - Arrigoni, Giorgio
AU - Cagnin, Stefano
AU - Biancotto, Giancarlo
AU - Franchin, Cinzia
AU - Sorgato, Maria Catia
AU - Bertoli, Alessandro
N1 - Funding Information:
Acknowledgements Authors are grateful to Drs. Domenico Azarnia Tehran and Fiorella Tonello for helpful suggestions on the synaptic vesicle recycling assay, and to Dr. Cesare Montecucco for kindly providing us with the needed serotype of botulinum toxin. This work was supported by the University of Padova (PRAT CPDA158035 to A.B. and CPDA139317 to S.C.). C.P. was supported by a grant to G.A. (CPDR159477/15). Authors also acknowledge the Cassa di Risparmio di Padova e Rovigo (Cariparo) Holding for funding acquisition of the LTQ-Orbitrap XL mass spectrometer.
Publisher Copyright:
© 2018, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2019/5/1
Y1 - 2019/5/1
N2 - The cellular prion protein (PrP C ), whose misfolded conformers are implicated in prion diseases, localizes to both the presynaptic membrane and postsynaptic density. To explore possible molecular contributions of PrP C to synaptic transmission, we utilized a mass spectrometry approach to quantify the release of glutamate from primary cerebellar granule neurons (CGN) expressing, or deprived of (PrP-KO), PrP C , following a depolarizing stimulus. Under the same conditions, we also tracked recycling of synaptic vesicles (SVs) in the two neuronal populations. We found that in PrP-KO CGN these processes decreased by 40 and 60%, respectively, compared to PrP C -expressing neurons. Unbiased quantitative mass spectrometry was then employed to compare the whole proteome of CGN with the two PrP genotypes. This approach allowed us to assess that, relative to the PrP C -expressing counterpart, the absence of PrP C modified the protein expression profile, including diminution of some components of SV recycling and fusion machinery. Subsequent quantitative RT-PCR closely reproduced proteomic data, indicating that PrP C is committed to ensuring optimal synaptic transmission by regulating genes involved in SV dynamics and neurotransmitter release. These novel molecular and cellular aspects of PrP C add insight into the underlying mechanisms for synaptic dysfunctions occurring in neurodegenerative disorders in which a compromised PrP C is likely to intervene.
AB - The cellular prion protein (PrP C ), whose misfolded conformers are implicated in prion diseases, localizes to both the presynaptic membrane and postsynaptic density. To explore possible molecular contributions of PrP C to synaptic transmission, we utilized a mass spectrometry approach to quantify the release of glutamate from primary cerebellar granule neurons (CGN) expressing, or deprived of (PrP-KO), PrP C , following a depolarizing stimulus. Under the same conditions, we also tracked recycling of synaptic vesicles (SVs) in the two neuronal populations. We found that in PrP-KO CGN these processes decreased by 40 and 60%, respectively, compared to PrP C -expressing neurons. Unbiased quantitative mass spectrometry was then employed to compare the whole proteome of CGN with the two PrP genotypes. This approach allowed us to assess that, relative to the PrP C -expressing counterpart, the absence of PrP C modified the protein expression profile, including diminution of some components of SV recycling and fusion machinery. Subsequent quantitative RT-PCR closely reproduced proteomic data, indicating that PrP C is committed to ensuring optimal synaptic transmission by regulating genes involved in SV dynamics and neurotransmitter release. These novel molecular and cellular aspects of PrP C add insight into the underlying mechanisms for synaptic dysfunctions occurring in neurodegenerative disorders in which a compromised PrP C is likely to intervene.
KW - Gene expression
KW - Mass spectrometry
KW - Neurotransmission
KW - Prion protein
KW - Selected reaction monitoring
KW - Synaptic vesicle
UR - http://www.scopus.com/inward/record.url?scp=85051804725&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85051804725&partnerID=8YFLogxK
U2 - 10.1007/s12035-018-1293-4
DO - 10.1007/s12035-018-1293-4
M3 - Article
C2 - 30128651
AN - SCOPUS:85051804725
SN - 0893-7648
VL - 56
SP - 3420
EP - 3436
JO - Molecular Neurobiology
JF - Molecular Neurobiology
IS - 5
ER -