Presynaptic localization of Smn and hnRNP R in axon terminals of embryonic and postnatal mouse motoneurons.

Benjamin Dombert,Michael Sendtner,Rajeeve Sivadasan,Christian M Simon,Sibylle Jablonka,Michael A Fox

doi:10.1371/journal.pone.0110846

Benjamin Dombert, Michael Sendtner + Show 4 more

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https://doi.org/10.1371/journal.pone.0110846

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Journal: PloS one	Publication Date: Oct 22, 2014
Citations: 58	License type: CC BY 4.0

Affiliation: University Hospital Würzburg

Abstract

Spinal muscular atrophy (SMA) is caused by deficiency of the ubiquitously expressed survival motoneuron (SMN) protein. SMN is crucial component of a complex for the assembly of spliceosomal small nuclear ribonucleoprotein (snRNP) particles. Other cellular functions of SMN are less characterized so far. SMA predominantly affects lower motoneurons, but the cellular basis for this relative specificity is still unknown. In contrast to nonneuronal cells where the protein is mainly localized in perinuclear regions and the nucleus, Smn is also present in dendrites, axons and axonal growth cones of isolated motoneurons in vitro. However, this distribution has not been shown in vivo and it is not clear whether Smn and hnRNP R are also present in presynaptic axon terminals of motoneurons in postnatal mice. Smn also associates with components not included in the classical SMN complex like RNA-binding proteins FUS, TDP43, HuD and hnRNP R which are involved in RNA processing, subcellular localization and translation. We show here that Smn and hnRNP R are present in presynaptic compartments at neuromuscular endplates of embryonic and postnatal mice. Smn and hnRNP R are localized in close proximity to each other in axons and axon terminals both in vitro and in vivo. We also provide new evidence for a direct interaction of Smn and hnRNP R in vitro and in vivo, particularly in the cytosol of motoneurons. These data point to functions of SMN beyond snRNP assembly which could be crucial for recruitment and transport of RNA particles into axons and axon terminals, a mechanism which may contribute to SMA pathogenesis.

Highlights

Proximal spinal muscular atrophy (SMA), the most common form of motoneuron disease in children and young adult, is caused by deficiency or loss of function of survival motoneuron (SMN) [1]
Smn is found in nuclear structures called Gemini of coiled bodies (Gems) where spliceosomal U small nuclear ribonucleoprotein (snRNP) are regenerated [17]
Localization of Smn and hnRNP R in spinal motoneurons and neuromuscular endplates Based on these results we studied distribution and colocalization of Smn and hnRNP R in spinal cord cross sections from E18 mouse embryos (Fig. 5A) which correlates with the developmental stage of primary motoneurons isolated at E13.5 and cultured for 5DIV

Summary

Introduction

Proximal spinal muscular atrophy (SMA), the most common form of motoneuron disease in children and young adult, is caused by deficiency or loss of function of SMN [1]. Recent studies with fly models have provided evidence that defects of pre-RNA splicing in sensory neurons contribute to the pathogenesis of SMA [3]. In SMA patients, motoneurons are primarily affected. Other organs and even most other types of neurons in the central and peripheral nervous system are spared or much less affected, raising the question about the molecular mechanisms underlying the high vulnerability of motoneurons. Weakness of the proximal musculature is an early feature in SMA patients and this correlates with defects in neurotransmission at neuromuscular junctions (NMJ) [9,10]

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