Presynaptic Biogenesis Requires Axonal Transport of Lysosome-Related Vesicles

Vukoja, Anela; Rey, Ulises; Petzoldt, Astrid G.; Ott, Christoph; Vollweiter, Dennis; Quentin, Christine; Puchkov, Dmytro; Reynolds, Eric C.; Lehmann, Martin; Hohensee, Svea; Rosa, Stefanie; Lipowsky, Reinhard; Sigrist, Stephan J.; Haucke, Volker

doi:10.1016/j.neuron.2018.08.004

Cited by 119 publications

(177 citation statements)

References 74 publications

(128 reference statements)

Supporting

Mentioning

156

Contrasting

Unclassified

Order By: Relevance

“…Also, transcripts coding for membrane proteins including endoplasmic reticulum proteins, such as Ergic1, Calr or Sec62, were diminished in the vGLUT1 + sorted synaptosomes. There was also a clear depletion of transcripts coding for integral synaptic vesicle proteins ( Fig 4D,F), consistent with a recent report of somatic vesicle biogenesis and transport (31). Of note, among the 468 transcripts enriched in vGLUT1 + terminals, 62 were known targets of the RNA-binding Fragile X Mental Retardation Protein (FMRP), the loss of which causes fragile X syndrome ( Fig.…”

Section: The Presynaptic Transcriptome Of Vglut1+ Terminalssupporting

confidence: 90%

Local protein synthesis is a ubiquitous feature of neuronal pre- and postsynaptic compartments

Hafner

Donlin-Asp

Leitch

et al. 2018

Preprint

116

View full text Add to dashboard Cite

Local protein synthesis is a ubiquitous feature of neuronal pre-and postsynaptic compartments.One sentence summary: Protein synthesis occurs in all synaptic compartments, including excitatory and inhibitory axon terminals. Presynaptic terminals from mouse cortex and hippocampus contain translation machineryEfforts to localize molecules or cell biological events to neuronal pre-or postsynaptic compartments using fluorescence microscopy are limited by the tight association of the axonal bouton and the dendritic spine or synapse; the synaptic cleft, the only clear region of separation, is only ~ 20 nm wide. Here, in order to increase the resolving power to visualize mRNA molecules in pre-and postsynaptic compartments, we optimized fluorescence in situ hybridization (FISH) and nascent protein detection methods for use with expansion microscopy (22) ( Fig. 1A; see Methods). We used adult mouse brain slices or rat cultured hippocampal neurons and found that expansion resulted in an enlargement of both pre-and postsynaptic compartments, with an average expansion of ~3.4 fold. This yielded a clear separation between the pre-and postsynaptic compartments. To evaluate whether ribosomes and mRNA species are present in defined presynaptic compartments, we used immunolabelling for either excitatory (vGLUT1; (23, 24) or inhibitory (vGAT; (25, 26)) nerve terminals in expanded mouse brain sections (both cortex and hippocampus) ( Fig. 1B-E) or rat cultured hippocampal neurons. We took care to identify the molecules-of-interest within individual z-sections positively labeled for excitatory or inhibitory terminals. We noted that signal detected outside of immunolabeled compartments corresponded to signal arising from nearby unlabeled cells. We detected ribosomes in a large majority (>75%) of both excitatory and inhibitory presynaptic nerve terminals, using antibodies directed against either a small (RPS11) or a large (RPL26) ribosomal protein ( Fig. 1 B-E). Next, we used FISH probes to detect 18s and 28s rRNA as well as polyadenylated mRNA (detected with a poly d(T) probe) in expanded samples ( Fig. 1B-E). Consistent with the abundance of ribosomal proteins, we detected rRNA in over 80% of both excitatory and inhibitory nerve terminals ( Fig. 1B-E). RNase treatment effectively reduced all rRNA signal. In cultured neurons, we also noted that poly(A) mRNA was abundant, as expected, in dendritic spines. In addition, we used an anti-tau antibody to label axons and detected both 18s and 28s rRNA in axonal segments. Thus, mRNAs and ribosomes were abundant in excitatory and inhibitory presynaptic nerve terminals from both mouse brain slices and rat hippocampal cultured neurons.

show abstract

Section: The Presynaptic Transcriptome Of Vglut1+ Terminalssupporting

confidence: 90%

Local protein synthesis is a ubiquitous feature of neuronal pre- and postsynaptic compartments

Hafner

Donlin-Asp

Leitch

et al. 2018

Preprint

116

View full text Add to dashboard Cite

show abstract

“…We found that activity-dependent MT nucleation at boutons is required for interbouton SV motility, and to allow neurotransmitter release (Figures 3 and 4). Arl8b facilitates the Kif1A mediated motility of both SVs and PLVs, which coordinate the anterograde transport of packets of AZ and SV proteins in developing neurons during presynaptic biogenesis (30,31,35). In addition, Kif1A was recently implicated in synaptic cargo anterograde delivery to en passant boutons using dynamic MTs as tracks (23).…”

Section: Discussionmentioning

confidence: 99%

Activity-dependent nucleation of dynamic microtubules at presynaptic boutons is required for neurotransmission

Kumar

Blockus

et al. 2019

Preprint

View full text Add to dashboard Cite

Highlights:• Excitatory boutons are hotspots for neuronal activity-induced γ-tubulin dependent MT nucleation• The augmin complex is required for the correct polarity of presynaptic de novo nucleated MTs • Presynaptic MT nucleation promotes SV motility and exocytosis at sites of release In Brief: Our results demonstrate that excitatory en passant boutons are hotspots for neuronal activity-induced γ-tubulin-and augmin-dependent oriented MT nucleation, and that the resulting presynaptic de novo nucleated MTs promote inter-bouton SV motility which is rate-limiting for neurotransmitter release. 2 SUMMARY Control of microtubule (MT) dynamics is critical for neuronal function. Whether MT nucleation is regulated at presynaptic boutons and influences overall presynaptic activity remains unknown. By visualizing MT dynamics at individual excitatory en passant boutons in axons of hippocampal neurons we found that MTs preferentially grow from presynaptic boutons as a result of g-tubulin and augmin-dependent nucleation. MT nucleation at boutons is promoted by neuronal activity, functionally coupled to synaptic vesicle (SV) transport, and required for neurotransmission. Hence, en passant boutons act as hotspots for activitydependent MT nucleation, which is required for neurotransmission by providing the tracks for a rate-limiting supply of SVs to sites of neurotransmitter release.

show abstract

“…Significantly, all three mRNAs are high confidence targets of FMRP in granule neurons ( Table 2). It is thought that the coordinated delivery of these active zone components to presynaptic domains by anterograde transport along microtubules using kinesin motors (including Kif1a) allows for their growth [169][170][171][172] . Notably, the top ranked mRNA target of FMRP in granule cells is Rapgef4 which encodes Epac2/RapgefII, a guanine nucleotide exchange factor for Rap1 and a sensor of cAMP levels, which are altered in both the mouse model and human patient cells [173][174][175][176] [190][191][192][193] and Emily Osterweil and colleagues have identified lovastatin as an ERK inhibitor which has shown efficacy in mouse 130 and rat 132 FXS models.…”

Section: Loss Of Function Of Fmrp Causes Altered Behavior and Cognitimentioning

confidence: 99%

FMRP binding to a ranked subset of long genes is revealed by coupled CLIP and TRAP in specific neuronal cell types

Driesche

Sawicka

Zhang

et al. 2019

Preprint

View full text Add to dashboard Cite

Highlights 1. We have created a mouse model in which endogenous FMRP can be conditionally tagged.2. Using tag-specific CLIP we describe ranked and specific sets of in vivo FMRP mRNA targets in two types of neurons.3. This ranking was used to reveal that FMRP regulates mRNAs with long coding sequences.4. FMRP mRNA targets in Purkinje cells, including the top-ranked IP3 receptor, are related to cell-autonomous Fragile X phenotypes. 5.We have updated our previous list of whole mouse brain FMRP mRNA targets with more replicates, deeper sequencing and improved analysis 6. The use of tagged FMRP in less abundant cell populations allowed identification of novel mRNA targets missed in a whole brain analysis

show abstract

Presynaptic Biogenesis Requires Axonal Transport of Lysosome-Related Vesicles

Cited by 119 publications

References 74 publications

Local protein synthesis is a ubiquitous feature of neuronal pre- and postsynaptic compartments

Local protein synthesis is a ubiquitous feature of neuronal pre- and postsynaptic compartments

Activity-dependent nucleation of dynamic microtubules at presynaptic boutons is required for neurotransmission

FMRP binding to a ranked subset of long genes is revealed by coupled CLIP and TRAP in specific neuronal cell types

Contact Info

Product

Resources

About