TY - JOUR
T1 - Plasma membrane calcium ATPase-neuroplastin complexes are selectively stabilized in GM1-containing lipid rafts
AU - Ilic, Katarina
AU - Lin, Xiao
AU - Malci, Ayse
AU - Stojanović, Mario
AU - Puljko, Borna
AU - Rožman, Marko
AU - Vukelić, Željka
AU - Heffer, Marija
AU - Montag, Dirk
AU - Schnaar, Ronald L.
AU - Kalanj-Bognar, Svjetlana
AU - Herrera-Molina, Rodrigo
AU - Mlinac-Jerkovic, Kristina
N1 - Funding Information:
The research was funded by Croatian Science Foundation grant NeuroReact, IP-2016-06-8636 (to SK-B), grant Raft tuning, IP-2014-09-2324 (to MH), and by German Academic Exchange Service (DAAD) n◦ 57514679 to KM-J and RH-M. KM-J gratefully acknowledges the support from EMBO (Short-term fellowship, ASTF 363-2015). KI gratefully acknowledges the support from Collaborative Research Center 779 “Neurobiology of Motivated Behavior” project to Dr. Karl-Heinz Smalla, Magdeburg, Germany. RH m acknowledges the grant from Center for Behavioral Brain Sciences (CBBS). RLS acknowledges support from the US National Institutes of Health (CA241953). DM acknowledges the support from German Federal Ministry of Education and Research (BMBF grant CONICYT to Eckart D. Gundelfinger, Karl-Heinz Smalla, Constanze Seidenbecher, and DM). AM is a fellow of the ABINEP graduate school (ESF, 2014–2020). XL is a fellow of the China Scholarship Council (n◦ 201506290028). This publication was co-financed by the European Union through the European Regional Development Fund, Operational Programme Competitiveness and Cohesion, grant agreement No. KK.01.1.1.01.0007 (CoRE –Neuro). Graphical abstract and Figure 5 were created with BioRender.com (Agreement number: ZA239UMOJD, last accessed 3 September 2021).
Funding Information:
Funding: The research was funded by Croatian Science Foundation grant NeuroReact, IP-2016-06-8636 (to SK-B), grant Raft tuning, IP-2014-09-2324 (to MH), and by German Academic Exchange Service (DAAD) n◦ 57514679 to KM-J and RH-M. KM-J gratefully acknowledges the support from EMBO (Short-term fellowship, ASTF 363-2015). KI gratefully acknowledges the support from Collaborative Research Center 779 “Neurobiology of Motivated Behavior” project to Dr. Karl-Heinz Smalla, Magdeburg, Germany. RH m acknowledges the grant from Center for Behavioral Brain Sciences (CBBS). RLS acknowledges support from the US National Institutes of Health (CA241953). DM acknowledges the support from German Federal Ministry of Education and Research (BMBF grant CONICYT to Eckart D. Gundelfinger, Karl-Heinz Smalla, Constanze Seidenbecher, and DM). AM is a fellow of the ABINEP graduate school (ESF, 2014–2020). XL is a fellow of the China Scholarship Council (n◦ 201506290028). This publication was co-financed by the European Union through the European Regional Development Fund, Operational Programme Competitiveness and Cohesion, grant agreement No. KK.01.1.1.01.0007 (CoRE –Neuro). Graphical abstract and Figure 5 were created with BioRender.com (Agreement number: ZA239UMOJD, last accessed 3 September 2021).
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/12/1
Y1 - 2021/12/1
N2 - The recent identification of plasma membrane (Ca2+)-ATPase (PMCA)-Neuroplastin (Np) complexes has renewed attention on cell regulation of cytosolic calcium extrusion, which is of particular relevance in neurons. Here, we tested the hypothesis that PMCA-Neuroplastin complexes exist in specific ganglioside-containing rafts, which could affect calcium homeostasis. We analyzed the abundance of all four PMCA paralogs (PMCA1-4) and Neuroplastin isoforms (Np65 and Np55) in lipid rafts and bulk membrane fractions from GM2/GD2 synthase-deficient mouse brains. In these fractions, we found altered distribution of Np65/Np55 and selected PMCA isoforms, namely PMCA1 and 2. Cell surface staining and confocal microscopy identified GM1 as the main complex ganglioside co-localizing with Neuroplastin in cultured hippocampal neurons. Furthermore, blocking GM1 with a specific antibody resulted in delayed calcium restoration of electrically evoked calcium transients in the soma of hippocampal neurons. The content and composition of all ganglioside species were unchanged in Neuroplastin-deficient mouse brains. Therefore, we conclude that altered composition or disorganization of ganglioside-containing rafts results in changed regulation of calcium signals in neurons. We propose that GM1 could be a key sphingolipid for ensuring proper location of the PMCA-Neuroplastin complexes into rafts in order to participate in the regulation of neuronal calcium homeostasis.
AB - The recent identification of plasma membrane (Ca2+)-ATPase (PMCA)-Neuroplastin (Np) complexes has renewed attention on cell regulation of cytosolic calcium extrusion, which is of particular relevance in neurons. Here, we tested the hypothesis that PMCA-Neuroplastin complexes exist in specific ganglioside-containing rafts, which could affect calcium homeostasis. We analyzed the abundance of all four PMCA paralogs (PMCA1-4) and Neuroplastin isoforms (Np65 and Np55) in lipid rafts and bulk membrane fractions from GM2/GD2 synthase-deficient mouse brains. In these fractions, we found altered distribution of Np65/Np55 and selected PMCA isoforms, namely PMCA1 and 2. Cell surface staining and confocal microscopy identified GM1 as the main complex ganglioside co-localizing with Neuroplastin in cultured hippocampal neurons. Furthermore, blocking GM1 with a specific antibody resulted in delayed calcium restoration of electrically evoked calcium transients in the soma of hippocampal neurons. The content and composition of all ganglioside species were unchanged in Neuroplastin-deficient mouse brains. Therefore, we conclude that altered composition or disorganization of ganglioside-containing rafts results in changed regulation of calcium signals in neurons. We propose that GM1 could be a key sphingolipid for ensuring proper location of the PMCA-Neuroplastin complexes into rafts in order to participate in the regulation of neuronal calcium homeostasis.
KW - B4galnt1
KW - GM2/GD2 synthase
KW - Gangliosides
KW - Glycosphingolipids
KW - Membrane microdomains
KW - Neuronal calcium homeostasis
UR - http://www.scopus.com/inward/record.url?scp=85121295305&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85121295305&partnerID=8YFLogxK
U2 - 10.3390/ijms222413590
DO - 10.3390/ijms222413590
M3 - Article
C2 - 34948386
AN - SCOPUS:85121295305
SN - 1661-6596
VL - 22
JO - International journal of molecular sciences
JF - International journal of molecular sciences
IS - 24
M1 - 13590
ER -