Dataset Information

Glycolytic enzyme aldolase A is necessary for reelin mediated neurite growth and arborization

ABSTRACT: Reelin is a large, secreted glycoprotein that is best known as an essential molecule directing neocortical neuronal migration, but reelin serves an important functional role in regulating neurite outgrowth and arborization as well. Canonically, reelin signals through the lipoprotein receptors apolipoprotein E receptor 2 (ApoER2) and very low-density lipoprotein receptor (VLDLR). Reelin can signal through non-canonical receptors and unidentified co-receptors as well, but the downstream effects of these non-canonical pathways are not well understood. Using an unbiased Tandem Mass Tag LC-MS/MS (TMT) proteomics screen and gene set enrichment analysis (GSEA), we identified overlapping and distinct intra-cellular pathways enriched downstream of canonical and non-canonical reelin signaling in primary murine neurons during periods of robust neurite arborization. We observed canonical signaling led to substantial changes in actin cytoskeletal dynamics and cell morphogenesis while non-canonical signaling preferentially regulated protein translation, suggesting reelin fine tunes neuronal structure and function through distinct signaling pathways. We also identified a novel node downstream of canonical reelin signaling, glycolytic enzyme and actin binding protein aldolase A (aldo A). Biochemical analysis revealed aldo A undergoes de novo translation and is dissociated from the actin cytoskeleton after acute reelin stimulation. Additionally, shRNA knockdown of aldo A in primary murine cultures and in vivo through in utero electroporation (IUE), reveal aldo A is necessary for reelin mediated neurite growth and the proper arborization and positioning of excitatory layer II/III neurons independent of glycolytic function. These results shed new light on the molecular mechanisms and signaling pathways regulating reelin dependent neuronal arborization.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Neuron, Cell Culture

DISEASE(S): Alzheimer's Disease

SUBMITTER: WEIWEI LIN

LAB HEAD: Andrew Emili

PROVIDER: PXD046667 | Pride | 2025-05-06

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20220622_NL_Gav_Prot_F1.raw	Raw
	20220622_NL_Gav_Prot_F10.raw	Raw
	20220622_NL_Gav_Prot_F11.raw	Raw
	20220622_NL_Gav_Prot_F12.raw	Raw
	20220622_NL_Gav_Prot_F2.raw	Raw

Items per page:

1 - 5 of 12

Publications

Beyond Glycolysis: Aldolase A Is a Novel Effector in Reelin-Mediated Dendritic Development.

Lagani Gavin D GD Sha Mingqi M Lin Weiwei W Natarajan Sahana S Kankkunen Marcus M Kistler Sabrina A SA Lampl Noah N Waxman Hannah H Harper Evelyn R ER Emili Andrew A Beffert Uwe U Ho Angela A

The Journal of neuroscience : the official journal of the Society for Neuroscience 20241016 42

Reelin, a secreted glycoprotein, plays a crucial role in guiding neocortical neuronal migration, dendritic outgrowth and arborization, and synaptic plasticity in the adult brain. Reelin primarily operates through the canonical lipoprotein receptors apolipoprotein E receptor 2 (Apoer2) and very low-density lipoprotein receptor (Vldlr). Reelin also engages with noncanonical receptors and unidentified coreceptors; however, the effects of which are less understood. Using high-throughput tandem mass ...[more]

PMID: 39227156

Similar Datasets

Project description:The APOE gene is diversified by three alleles e2, e3, and e4 encoding corresponding apolipoprotein (apo) E isoforms. The e4 allele promotes age-related cognitive decline, risk of Alzheimer’s disease (AD), and the rate of AD dementia progression. ApoE is produced by astrocytes as high-density lipoprotein-like particles and these are internalized by neurons upon binding to neuron-expressed apoE receptors. ApoE isoforms differentially engage neuronal plasticity through poorly understood mechanisms. We examined here the effects of native apoE lipoproteins produced by immortalized astrocytes homozygous for e2, e3, and e4 alleles on the maturation and the transcriptomic profile of primary hippocampal neurons. Control neurons were grown in the presence of conditioned media from Apoe-/- astrocytes. ApoE2 and apoE3 significantly increase the dendritic arbor branching, the combined neurite length, and the total arbor surface of the hippocampal neurons, while apoE4 fails to produce similar effects and even significantly reduces the combined neurite length compared to the control. ApoE lipoproteins show no systemic effect on dendritic spine density, yet apoE2 and apoE3 increase the mature spines fraction, while apoE4 increases the immature spine fraction. This is associated with opposing effects of apoE2 or apoE3 and apoE4 on the expression of NR1 NMDA receptor subunit and PSD95. There are 1,062 genes differentially expressed across neurons cultured in the presence of apoE lipoproteins compared to the control. KEGG enrichment and gene ontology analyses show common for apoE2 and apoE3 activation of genes involved in neurite branching, and synaptic signaling. In contrast, apoE4 cultured neurons show upregulation of genes related to the glycolipid metabolism, which are involved in dendritic spine turnover and those which are usually silent in neurons and are related to cell cycle and DNA repair. In conclusion our work reveals that lipoprotein particles comprised of various apoE isoforms differentially regulate neuronal development through interaction with neuronal transcriptome. ApoE4 produces a functionally distinct transcriptomic profile, which is associated with attenuated neuronal development. Differential regulation of neuronal transcriptome by apoE isoforms is a newly identified biological mechanism, which has both implication in the development and aging of the CNS.

Project description:Stem cells are a potential key strategy for treating neurodegenerative diseases in which the generation of new neurons is critical. A better understanding of the characteristics and molecular properties of neural stem cells (NSC) and differentiated neurons can help in assessing neuronal maturity and possibly in devising better therapeutic strategies. We have therefore performed an in-depth gene expression profiling study of the C17.2 NSC line and primary neurons (PN) derived from embryonic mouse brains. Microarray analysis revealed a neuron-specific gene expression signature that distinguishes PN from NSCs, with elevated levels of transcripts involved in neuronal functions such as neurite development, axon guidance, in PN. The same comparison revealed decreased levels of multiple cytokine transcripts such as IFN, TNF, TGF, and IL. Among the differentially expressed genes, we found a statistically significant enrichment of genes in the ephrin, neurotrophin, CDK5 and actin pathways which control multiple neuronal-specific functions. Furthermore, genes involved in cell cycle were among the most significantly changed in PN. In order to better understand the role of cell cycle arrest in mediating NSCs differentiation, we blocked the cell cycle of NSCs with Mitomycin C (MMC) and examined cellular morphology and gene expression signatures. Although these MMC-treated NSCs displayed a neuronal morphology and expressed some neuronal differentiation marker genes, their gene expression patterns was very different from primary neurons. We conclude that: 1) Fully differentiated primary neurons display a specific neuronal gene expression signature; 2) cell-cycle block in NSC does not induce the formation of fully differentiated neurons; 3) Cytokines such as IFN, TNF, TGF and IL are part of normal NSC function and/or physiology; 4) Signaling pathways of ephrin, neurotrophin, CDK5 and actin, related to major neuronal features, are dynamically enriched in genes showing changes in expression level. Gene expression profiles in neuronal stem cell, mitomycin-treated neuronal stem cells and primary neuronal cultures were compared to examine cellular morphology and gene expression signatures during neuronal differentiation.