bioRxiv Science⌕ Search

Biology subjects

Pauzin, F. P.

Publications and source records attributed to Pauzin, F. P..

6 recordsLinked to original sources

Tactile responses in the mouse perirhinal cortex show invariance to physical features of the stimulus

Sensory processing in the cortex follows a hierarchical structure, transforming fine-grained stimulus features into abstract, invariant representations. These transformations have typically been characterised in visual pathways ranging from primary visual cortex to inferotemporal cortex. Less is known about how stimulus representations are formatted in other modalities and downstream regions such as the perirhinal cortex (PER), which receives processed sensory input from all modalities and is involved in object recognition. To address this, we investigated neural activity along the somatosensory hierarchy in awake, head-fixed mice receiving tactile whisker stimulation on either side of the face. PCA and stimulus-baseline decoding analyses revealed that PER activity was similar across stimulated sides, while this was not the case in primary somatosensory cortex (wS1). We also performed decoding analyses for the side of stimulation and the stimulus movement direction and found that wS1 and secondary somatosensory cortex could reliably decode both features. In contrast, PER was shown to be invariant to both features. We also clustered neurons based on their response dynamics, which indicated that the majority of PER neurons did not encode the basic stimulus features. These findings demonstrate that PER is a critical node in the somatosensory processing hierarchy, encoding tactile information in an invariant and abstract format. Together, these insights contribute to a more refined understanding of how perceptual processing in PER contributes to the various cognitive processes it is utilised for. Significance StatementSensory information is processed in stages that transform detailed signals into complex, stable representations. The perirhinal cortex (PER) is at the apex of these stages and is involved in recognizing objects, a cognitive process that requires both perceptual and memory-based processing. While PERs role in the latter is established, evidence for the former is conflicting. To investigate this, we recorded PER activity in mice passively experiencing simple tactile stimulation with two pairs of feature combinations. Along the cortical somatosensory hierarchy invariance of stimulus representation emerged in PER, while individual features were represented in earlier cortices with high fidelity. This suggests that PER contributes to the formation of invariant representations of sensory stimuli that is a foundation for object recognition.

neuroscience↗

Arc/Arg3.1 binds the nuclear polyadenylate-binding protein RRM and regulates neuronal activity-dependent formation of nuclear speckles

Arc is a neuronal activity-induced protein interaction hub with critical roles in synaptic plasticity and memory. Arc localizes to synapses and the nucleus, but its nuclear functions are little known. We show that Arc accumulates in the interchromatin space of dentate granule cell nuclei and the nucleosol subcellular fraction following seizure activity and in vivo dentate gyrus LTP. Proteomic analysis of affinity-purified Arc complexes identified proteins with functions in post-transcriptional mRNA processing. During LTP, Arc undergoes enhanced complex formation with polyadenylate binding protein nuclear 1 (PABPN1) and paraspeckle splicing factor (PSF) in the nucleosol. In vitro peptide binding arrays show selective binding of Arc to the PABPN1 polyA RNA recognition motif. In hippocampal neuronal cultures, Arc knockdown increases formation of PABPN1 nuclear speckles and blocks chemical-LTP associated increases in small PABPN1 foci. These results implicate Arc in basal and neuronal activity-dependent regulation of PABPN1 speckles involved in mRNA processing and polyadenylation.

neuroscience↗

Terminal nucleotidyltransferase Tent2 microRNA tailing regulates excitatory/inhibitory balance in the hippocampus

One of the post-transcriptional mechanisms regulating the stability of RNA molecules involves the addition of non-templated nucleotides to their 3 ends, a process known as RNA tailing. To systematically investigate the physiological consequences of terminal nucleotidyltransferase TENT2 absence on RNA 3 end modifications in the mouse hippocampus we developed a new Tent2 knockout mouse. Electrophysiological measurements revealed increased excitability in Tent2 KO hippocampal neurons, and behavioral analyses showed decreased anxiety and improved fear extinction in these mice. At the molecular level, we observed a significant contribution of TENT2 to the monoadenylation of various classes of miRNAs, but found no effect of the enzymes loss on the total poly(A) tail length of mRNAs, as measured by Direct Nanopore RNA sequencing. Alterations in monoadenylation of a large population of microRNAs affected the overall mRNA abundance, particularly transcripts related synaptic transmission, which were downregulated in the hippocampus of Tent2 knockout mice. These changes explain the observed behavioral and electrophysiological alterations. Our data thus establish a link between TENT2-dependent microRNA tailing and the balance of inhibitory and excitatory neurotransmission.

neuroscience↗

A nanobody-based proximity ligation assay detects constitutive and stimulus-regulated native Arc/Arg3.1 oligomers in hippocampal neuronal dendrites

Activity-regulated cytoskeleton-associated (Arc), the product of an immediate early gene, plays critical roles in synaptic plasticity and memory. Evidence suggests that Arc function is determined by its oligomeric state, however, methods for localization of native Arc oligomers are lacking. Here, we developed a nanobody-based proximity ligation assay (PLA) for detection, localization, and quantification of Arc-Arc complexes in primary rat hippocampal neuronal cultures. We used nanobodies with single, structurally defined epitopes in the bilobar Arc capsid domain. Nanobody H11 binds inside the N-lobe ligand pocket, while nanobody C11 binds to the C-lobe surface. For each nanobody, ALFA- and FLAG-epitope tags created a platform for antibody binding and PLA. Surprisingly, PLA puncta in neuronal dendrites revealed widespread constitutive Arc-Arc complexes. Treatment of cultures with tetrodotoxin or cycloheximide had no effect, suggesting stable complexes that are independent of recent neuronal activity and protein synthesis. To assess detection of oligomers, cultures were exposed to a cell-penetrating peptide inhibitor of the Arc oligomerization motif (OligoOFF). Arc-Arc complexes detected by H11 PLA were inhibited by OligoOff but not by control peptide. Notably, Arc complexes detected by C11 were unaffected by OligoOFF. Furthermore, we evaluated Arc complex formation after chemical stimuli that increase Arc synthesis. Brain-derived neurotrophic factor increased Arc-Arc signal detected by C11, but not H11. Conversely, dihydroxyphenylglycine (DHPG) treatment selectively enhanced H11 PLA signals. In sum, nanobody-based PLA reveals constitutive and stimulus-regulated Arc oligomers in hippocampal neuronal dendrites. A model is proposed based on detection of Arc dimer by C11 and higher-order oligomer by H11 nanobody.

neuroscience↗

The polyadenylation landscape after in vivo long-term potentiation in the rat brain

Local protein synthesis in neurons is vital for synaptic maintenance and plasticity, yet the regulatory mechanisms, particularly cytoplasmic polyadenylation, are not fully understood. This study employed nanopore sequencing to examine transcriptomic responses in rat hippocampi during in vivo long-term potentiation (LTP) and in synaptoneurosomes after in vitro stimulation. Our long-read transcriptomic dataset allows for detailed analysis of mRNA 3'-ends, poly(A) tail lengths, and nucleotide composition. We observed dynamic shifts in polyadenylation site preference post-LTP induction, with significant poly(A) tail lengthening restricted to transcriptionally induced mRNAs. The poly(A) tails of these genes showed increased non-adenosine abundance. In synaptoneurosomes, chemical stimulation led to shortening of poly(A) tails on preexisting mRNAs, indicating translation-induced deadenylation. Additionally, we discovered a group of neuronal transcripts with poly(A) tails abundant in non-adenosine residues. These tails are semi-templated and derived from extremely adenosine-rich 3'UTRs. This study provides a comprehensive overview of mRNA 3'-end dynamics during LTP, offering insights into post-transcriptional regulation following synaptic activation of plasticity in neurons.

neuroscience↗

Trunk sensory and motor cortex ispreferentially integrated with hindlimb sensoryinformation that supports trunk stabilization

Sensorimotor integration in the trunk system has been poorly studied despite its importance for examining functional recovery after neurological injury or disease. Here, we mapped the relationship between thoracic dorsal root ganglia and trunk sensory cortex (S1) to create a detailed map of the extent and internal organization of trunk primary sensory cortex, and trunk primary motor cortex (M1) and showed that both cortices are somatotopically complex structures that are larger than previously described. Surprisingly, projections from trunk S1 to trunk M1 were not anatomically organized. We found relatively weak sensorimotor integration between trunk M1 and S1 and between trunk M1 and forelimb S1 compared to extensive integration between trunk M1 and hindlimb S1 and M1. This strong trunk/hindlimb connection was identified for high intensity stimuli that activated proprioceptive pathways. To assess the implication of this integration, the responses in sensorimotor cortex were examined during a postural control task and supported sensorimotor integration between hindlimb sensory and lower trunk motor cortex. Together, these data suggest that trunk M1 is guided predominately by hindlimb proprioceptive information that reached the cortex directly via the thalamus. This unique sensorimotor integration suggests an essential role for the trunk system in postural control, and its consideration could be important for understanding studies regarding recovery of function after spinal cord injury. SignificanceThis work identifies extensive sensorimotor integration between trunk and hindlimb cortices, demonstrating that sensorimotor integration is an operational mode of the trunk cortex in intact animals. The functional role of this integration was demonstrated for postural control when the animal was subjected to lateral tilts. Furthermore, these results provide insight into cortical reorganization after spinal cord injury making clear that sensorimotor integration after SCI is an attempt to restore sensorimotor integration that existed in the intact system. These results could be used to tailor rehabilitative strategies to optimize sensorimotor integration for functional recovery.

neuroscience↗