Phosphatidylinositol 3,5-bisphosphate facilitates axonal vesicle transport and presynapse assembly

Rizalar, Filiz Sila; Lucht, Max T.; Petzoldt, Astrid; Kong, Shuhan; Sun, Jiachen; Vines, James H.; Telugu, Narasimha Swamy; Diecke, Sebastian; Kaas, Thomas; Bullmann, Torsten; Schmied, Christopher; Löwe, Delia; King, Jason S.; Cho, Wonhwa; Hallermann, Stefan; Puchkov, Dmytro; Sigrist, Stephan J.; Haucke, Volker

doi:10.1126/science.adg1075

Cited by 14 publications

(3 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The transport of SV-lysosomes, like SVp carriers, also depends on UNC-104 (Figs 3B, S1H and S5A). Although our work has the limitation of using primarily overexpression-based transgenic lines to assess the composition of transport carriers, some of our observations in wild type neurons are corroborated by recent studies examining the trafficking of SVps in mammalian systems using CRISPR knock-in reporter lines [55].…”

Section: Discussionsupporting

confidence: 63%

LRK-1/LRRK2 and AP-3 regulate trafficking of synaptic vesicle precursors through active zone protein SYD-2/Liprin-α

Nadiminti,

Dixit,

Ratnakaran

et al. 2024

PLoS Genet

View full text Add to dashboard Cite

Synaptic vesicle proteins (SVps) are transported by the motor UNC-104/KIF1A. We show that SVps travel in heterogeneous carriers in C. elegans neuronal processes, with some SVp carriers co-transporting lysosomal proteins (SV-lysosomes). LRK-1/LRRK2 and the clathrin adaptor protein complex AP-3 play a critical role in the sorting of SVps and lysosomal proteins away from each other at the SV-lysosomal intermediate trafficking compartment. Both SVp carriers lacking lysosomal proteins and SV-lysosomes are dependent on the motor UNC-104/KIF1A for their transport. In lrk-1 mutants, both SVp carriers and SV-lysosomes can travel in axons in the absence of UNC-104, suggesting that LRK-1 plays an important role to enable UNC-104 dependent transport of synaptic vesicle proteins. Additionally, LRK-1 acts upstream of the AP-3 complex and regulates its membrane localization. In the absence of the AP-3 complex, the SV-lysosomes become more dependent on the UNC-104-SYD-2/Liprin-α complex for their transport. Therefore, SYD-2 acts to link upstream trafficking events with the transport of SVps likely through its interaction with the motor UNC-104. We further show that the mistrafficking of SVps into the dendrite in lrk-1 and apb-3 mutants depends on SYD-2, likely by regulating the recruitment of the AP-1/UNC-101. SYD-2 acts in concert with AP complexes to ensure polarized trafficking & transport of SVps.

show abstract

Section: Discussionsupporting

confidence: 63%

LRK-1/LRRK2 and AP-3 regulate trafficking of synaptic vesicle precursors through active zone protein SYD-2/Liprin-α

Nadiminti,

Dixit,

Ratnakaran

et al. 2024

PLoS Genet

View full text Add to dashboard Cite

show abstract

“…The hiPSC line BIHi005-A-24 (hPSCreg link: https://hpscreg.eu/cell-line/BIHi005-A-24 ) stably overexpressing the TetO-NGN2-T2A-PURO cassette was obtained from Berlin Institute of Health (BIH), Germany. This cell line constitutively expresses a Tet-ON activator which in turn allows induction of Ngn2 expression and the puromycin resistance gene by addition of doxycycline to the culture medium ( Rizalar et al, 2023 ). For capacitance measurements, we used the iPSC line NWTC11.G3.0036 ( Karch et al, 2019 ) derived from WTC11 (hPSCreg link: https://hpscreg.eu/cell-line/UCSFi001-A ), which also constitutively expresses a Tet-ON activator and was differentiated into glutamatergic neurons using the same protocol.…”

Section: Methodsmentioning

confidence: 99%

Human iPSC-Derived Neurons with Reliable Synapses and Large Presynaptic Action Potentials

Bullmann,

Kaas,

Ritzau-Jost

et al. 2024

J. Neurosci.

View full text Add to dashboard Cite

Understanding the function of the human brain requires determining basic properties of synaptic transmission in human neurons. One of the most fundamental parameters controlling neurotransmitter release is the presynaptic action potential, but its amplitude and duration remain controversial. Presynaptic action potentials have so far been measured with high temporal resolution only in a limited number of vertebrate but not in human neurons. To uncover properties of human presynaptic action potentials, we exploited recently developed tools to generate human glutamatergic neurons by transient expression of Neurogenin 2 (Ngn2) in pluripotent stem cells. During maturation for 3 to 9 weeks of culturing in different established media, the proportion of cells with multiple axon initial segments decreased, while the amount of axonal tau protein and neuronal excitability increased. Super-resolution microscopy revealed the alignment of the pre- and postsynaptic proteins, Bassoon and Homer. Synaptic transmission was surprisingly reliable at frequencies of 20, 50, and 100 Hz. The synchronicity of synaptic transmission during high-frequency transmission increased during 9 weeks of neuronal maturation. To analyze the mechanisms of synchronous high-frequency glutamate release, we developed direct presynaptic patch-clamp recordings from human neurons. The presynaptic action potentials had large overshoots to ∼25 mV and short durations of ∼0.5 ms. Our findings show that Ngn2-induced neurons represent an elegant model system allowing for functional, structural, and molecular analyses of glutamatergic synaptic transmission with high spatio-temporal resolution in human neurons. Furthermore, our data predict that glutamatergic transmission is mediated by large and rapid presynaptic action potentials in the human brain.Significance statementPresynaptic physiology remains poorly understood despite its relevance to neurological and psychiatric diseases. Studying presynaptic functions in human iPSC-derived neurons offers the important advantage of characterizing molecular mechanisms of neurotransmitter release in neurons derived from diseased patients. As a first step towards this goal, we established direct presynaptic whole-cell patch-clamp recordings from human glutamatergic neurons induced by transient Neurogenin 2 overexpression. We furthermore analyzed the structure of the synapses with super-resolution light microscopy and the synaptic short-term plasticity during high-frequency transmission. Our findings show that synchronous high-frequency transmission is mediated by rapid and large presynaptic action potentials in human neurons, similar to small conventional nerve terminals of rodent neurons.

show abstract

“…4(c) – 4(d) ]. Together with genome editing strategies to tag with SEP endogenous receptors or transporters, 16 , 39 – 41 these new approaches are paving the way to a detailed understanding of the trafficking steps at play in intact neuronal networks.…”

Section: Imaging and Manipulating Synaptic Functionmentioning

confidence: 99%

Understanding the nervous system: lessons from Frontiers in Neurophotonics

De Koninck,

Alonso,

Bancelin

et al. 2024

Neurophoton.

View full text Add to dashboard Cite

The Frontiers in Neurophotonics Symposium is a biennial event that brings together neurobiologists and physicists/engineers who share interest in the development of leading-edge photonics-based approaches to understand and manipulate the nervous system, from its individual molecular components to complex networks in the intact brain. In this Community paper, we highlight several topics that have been featured at the symposium that took place in October 2022 in Québec City, Canada.

show abstract

Phosphatidylinositol 3,5-bisphosphate facilitates axonal vesicle transport and presynapse assembly

Cited by 14 publications

References 68 publications

LRK-1/LRRK2 and AP-3 regulate trafficking of synaptic vesicle precursors through active zone protein SYD-2/Liprin-α

LRK-1/LRRK2 and AP-3 regulate trafficking of synaptic vesicle precursors through active zone protein SYD-2/Liprin-α

Human iPSC-Derived Neurons with Reliable Synapses and Large Presynaptic Action Potentials

Understanding the nervous system: lessons from Frontiers in Neurophotonics

Contact Info

Product

Resources

About