SULTA4A1 modulates synaptic development and function by promoting the formation of PSD-95/NMDAR complex
Lorenza Culotta,Benedetta Terragni,Ersilia Vinci,Alessandro Sessa,Vania Broccoli,Massimo Mantegazza,Chiara Verpelli +6 more
TL;DR: It is demonstrated that the pharmacological inhibition of Pin1 reverses the pathological phenotypes of SULT4A1 knockdown neurons by specifically restoring dendritic spine density and rescuing NMDAR-mediated synaptic transmission.
read more
Abstract: Sulfotransferase 4A1 (SULT4A1) is a cytosolic sulfotransferase, that is highly conserved across species and extensively expressed in the brain. However, the biological function of SULT4A1 is unclear. SULT4A1 has been implicated in several neuropsychiatric disorders, such as Phelan-McDermid Syndrome and schizophrenia. Here, we investigate the role of SULT4A1 within neuron development and function. Our data demonstrate that SULT4A1 modulates neuronal branching complexity and dendritic spines formation. Moreover, we show that SULT4A1, by negatively regulating the catalytic activity of Pin1 towards PSD-95, facilitates NMDAR synaptic expression and function. Finally, we demonstrate that the pharmacological inhibition of Pin1 reverses the pathological phenotypes of SULT4A1 knockdown neurons by specifically restoring dendritic spine density and rescuing NMDAR-mediated synaptic transmission. Together, these findings identify SULT4A1 as a novel player in neuron development and function by modulating dendritic morphology and synaptic activity.
read more
Chat with Paper
AI Agents for this Paper
Find similar papers on Google Scholar, PubMed and Arxiv
Write a critical review of this paper
Analyze citations of this paper to find unaddressed research gaps
Figures
Figure 3 │SULT4A1 knockdown alters synaptic transmission. (a) Representative immunoblots of total protein lysates derived from day-in-vitro 14 rat cortical neurons transduced with a lentivirus expressing shCtrl or shSULT4A1. Both conditions were compared with not infected condition (Ni) so to verify whether the infection itself could cause any alteration of SULT4A1 protein expression. SULT4A1 protein levels were normalized against the level of the not infected neurons (Ni n=7, shCtrl n=4, shSULT4A1 n=7; One-sample t test, **** P<0.0001, ns= not significant). (b) Representative western blots of total lysates from shCtrl- or shSULT4A1-transduced cortical neurons. Protein levels were normalized against the level of the shCtrl-transduced neurons. (shCtrl n≥3, shSULT4A1 n≥3; One-sample t test, * P<0.05). (c) Left: representative spontaneous excitatory postsynaptic currents (sEPSCs) recorded from shCtrl- (blue) or shSULT4A1- (red) transfected neurons. Center: plot of cumulative probability of currents frequency (shCtrl n=5, shSULT4A1 n=5; Kolmogorov-Smirnov test, P=4.16x10 -108 ). Right: mean instantaneous frequencies plot (shCtrl n=5, shSULT4A1 n=5; Mann-Whitney test, * P<0.05). (d) Left: representative spontaneous inhibitory postsynaptic currents (sIPSCs) recorded from shCtrl- (blue) or shSULT4A1- (red) transfected neurons. Center: plot of cumulative probability of currents frequency (shCtrl n=5, shSULT4A1 n=6; Kolmogorov-Smirnov test, P=4.85x10 -8 ). Right: mean instantaneous frequencies plot (shCtrl n=5, shSULT4A1 n=6; Mann-Whitney test). Data represent Mean ± SEM. 
Figure 6 │SULT4A1 modulates NMDAR synaptic expression and function. SULT4A1 interacts with Pin1 promoting the formation of the PSD-95/NMDAR complex in excitatory synapses. In absence of SULT4A1, Pin1 is free to interact with PSD-95 reducing NMDAR expression in synapses. 
Figure 5 │Partial rescue of shSULT4A1-dependent phenotype via Pin1 pharmacological 
Figure 4 │SULT4A1 interaction with Pin1 and its role in excitatory synapses. (a) Left: representative NMDA-mediated current (INMDA) traces elicited by the perfusion of 100 µM NMDA from shCtrl- (blue) and shSULT4A1- (red) transfected neurons. Right: analysis of mean peak INMDA current densities in shCtrl- and shSULT4A1-transfected neurons (shCtrl n=9, shSULT4A1 n=13; Unpaired t test, * P<0.05). (b) Representative western blots of synaptosomal fractions obtained from cortical neurons transduced with shCtrl or shSULT4A1. Protein levels were normalized against the levels of the shCtrl-transduced neurons (shCtrl n≥3, shSULT4A1 n≥3; One-sample t test, * P<0.05, ** P<0.01, *** P<0.001). (c) Left: representative images of immunoprecipitation (IP) assay performed on synaptosomal fractions derived from neurons transduced with shCtrl or shSULT4A1. Proteins were precipitated using mouse anti-Pin1 or mouse IgG antibodies and nitrocellulose membranes were probed with anti-Pin1, anti-SULT4A1 and anti-PSD-95 antibodies (WB). Right: histogram showing the ratio between PSD-95 and Pin1 signals (PSD-95/Pin1) (shCtrl n=4, shSULT4A1 n=4; Unpaired t test, * P<0.05). Data represent Mean ± SEM. 
Figure 1 figure supplement 1│Physiological expression of SULT4A1 in primary cortical neurons and iPSC-derived neurons. (a) Representative immunocytochemical staining for SULT4A1 (green), βIII-Tubulin (red) and DAPI (blue), in rat cortical neurons at day-in-vitro (DIV) 1, 7 and 14. (b) Left: representative immunocytochemical staining of Oct4 (green), Sox2 (red) and DAPI (blue) of induced pluripotent stem cells (iPSCs) obtained from healthy control individuals. Center: iPSC-derived neural stem cells (NSCs) stained for Nestin (green), Sox2 (red) and DAPI (blue). Right: representative immunostaining for MAP2 (green) and DAPI (blue) of NSC-derived neurons after 40 days of differentiation. Scale bar = 50μm. 
Figure 1 figure supplement 1│Physiological expression of SULT4A1 in primary cortical neurons and iPSC-derived neurons. (a) Representative immunocytochemical staining for SULT4A1 (green), βIII-Tubulin (red) and DAPI (blue), in rat cortical neurons at day-in-vitro (DIV) 1, 7 and 14. (b) Left: representative immunocytochemical staining of Oct4 (green), Sox2 (red) and DAPI (blue) of induced pluripotent stem cells (iPSCs) obtained from healthy control individuals. Center: iPSC-derived neural stem cells (NSCs) stained for Nestin (green), Sox2 (red) and DAPI (blue). Right: representative immunostaining for MAP2 (green) and DAPI (blue) of NSC-derived neurons after 40 days of differentiation. Scale bar = 50μm.
Citations
Sulfotransferase 4A1 activity facilitates sulfate-dependent cellular protection to oxidative stress
TL;DR: SULT4A1 has been assessed through all classical SULT approaches yet no SULT activity has been reported as discussed by the authors , however, they utilized Saccharomyces cerevisiae as a model system, which exhibits no endogenous SULT-related genes.
Sulfation Pathways During Neurodevelopment
TL;DR: In this article , the authors curated the temporal and spatial expression of sulfate maintenance genes in human fetal brain from 4 to 17 weeks post conception using the online Human Developmental Biology Resource Expression.
References
In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector
Luigi Naldini,Ulrike Blömer,Philippe Gallay,Daniel S. Ory,Richard C. Mulligan,Fred H. Gage,Inder M. Verma,Didier Trono +7 more
TL;DR: The ability of HIV-based viral vectors to deliver genes in vivo into nondividing cells could increase the applicability of retroviral vectors in human gene therapy.
5.4K
Germline Transmission and Tissue-Specific Expression of Transgenes Delivered by Lentiviral Vectors
TL;DR: Transgenic mice carrying the green fluorescent protein (GFP) gene driven by a ubiquitously expressing promoter are generated and transgenic rats that express GFP at high levels are generated, suggesting that this technique can be used to produce other transgenic animal species.
2.2K
Mutations in the gene encoding the synaptic scaffolding protein SHANK3 are associated with autism spectrum disorders.
Christelle M. Durand,Catalina Betancur,Tobias M. Boeckers,Juergen Bockmann,Pauline Chaste,Fabien Fauchereau,Gudrun Nygren,Maria Råstam,I. Carina Gillberg,Henrik Anckarsäter,Eili Sponheim,Hany Goubran-Botros,Richard Delorme,Nadia Chabane,Marie-Christine Mouren-Simeoni,Philippe de Mas,Eric Bieth,Bernadette Rogé,Delphine Héron,Lydie Burglen,Christopher Gillberg,Marion Leboyer,Thomas Bourgeron +22 more
TL;DR: It is reported that a mutation of a single copy of SHANK3 on chromosome 22q13 can result in language and/or social communication disorders.
1.5K
Shank3 mutant mice display autistic-like behaviours and striatal dysfunction
João Peça,Catia Feliciano,Jonathan T. Ting,Wenting Wang,Michael F. Wells,Talaignair N. Venkatraman,Christopher D. Lascola,Zhanyan Fu,Zhanyan Fu,Guoping Feng,Guoping Feng,Guoping Feng +11 more
TL;DR: In this paper, Shank3 gene deletions were found to lead to repetitive grooming and impaired social interaction in mice with autism spectrum disorders (ASDs) and other non-syndromic ASDs.
Shank, a Novel Family of Postsynaptic Density Proteins that Binds to the NMDA Receptor/PSD-95/GKAP Complex and Cortactin
Scott Naisbitt,Eunjoon Kim,Jian Cheng Tu,Bo Xiao,Carlo Sala,Juli G. Valtschanoff,Richard J. Weinberg,Paul F. Worley,Morgan Sheng +8 more
TL;DR: A novel family of postsynaptic density proteins, termed Shank, that binds via its PDZ domain to the C terminus of PSD-95-associated protein GKAP, and may function as a scaffold protein in the PSD, potentially cross-linking NMDA receptor/PSD- 95 complexes and coupling them to regulators of the actin cytoskeleton.
1K