Translational and Posttranslational Dynamics in a Model Peptidergic System

Bárez-López, Soledad; Mecawi, André Souza; Bryan, Natasha; Pauža, Audrys G; Duque, Victor J.; Gillard, Benjamin; Murphy, David; Greenwood, Michael

doi:10.1016/j.mcpro.2023.100544

Cited by 2 publications

(4 citation statements)

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“…With the dorsal/ventral distribution present in both the female and male rats in our study, this could potentially be the first demonstration of this anatomic distribution of Avp and Oxt neurons in the SON of both sexes using spatial transcriptomics. Our current data were also significantly correlated with the results from SON transcriptomics data from bulk RNAseq [ 19 ] and a separate proteomics study [ 34 ].…”

Section: Discussionsupporting

confidence: 76%

“…In addition to comparing the number of genes identified by both DE analysis methods, we compared the Visium spatial transcriptomics data to previously published SON transcriptomic and proteomic data sets. The relationships among the spatial transcriptomics data from the male rats ( n = 4) and previously published SON bulk RNAseq [ 19 ] and Proteome [ 34 ] data from normal male rats are illustrated in Fig. 3 D. Significant positive correlations were observed for the relationships between Visium SON transcriptomics data and bulk SON RNAseq data (Spearman r = 0.726, p < 0.001; Fig.…”

Section: Resultsmentioning

confidence: 67%

“…When rats are stimulated by systemic hyperosmolality, such as chronic salt loading and water deprivation [ 19 , 47 ], the expression of Caprin2 is highly increased in the SON. In rats submitted to hyperosmolality induced by water deprivation, Caprin2 is strongly hyperphosphorylated in the SON [ 34 ] and its expression increases in both AVP and OXT MNCs [ 48 ]. Knockdown of Caprin2 in the SON of dehydrated male rats attenuated the normal increase in AVP mRNA but paradoxically was also associated with increased plasma AVP [ 46 ].…”

Section: Discussionmentioning

confidence: 99%

“…To determine how the conservative and less conservative Loupe Browse analyses output correlates with the DESeq2 outcome, we performed a Spearman correlation analysis between the Loupe Browse gene count average with the DESeq2 normalized count average, considering all males and females for both cases. Next, using published micro-punched SON bulk RNAseq [ 19 ] and Proteome [ 34 ] data, we also performed Spearman correlation analyses to compare the Visium-identified basal male rat SON genes with genes and proteins previously identified in the male rat SON.…”

Section: Methodsmentioning

confidence: 99%

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Spatial transcriptomics reveal basal sex differences in supraoptic nucleus gene expression of adult rats related to cell signaling and ribosomal pathways

Nguyen,

Duque,

Phillips

et al. 2023

Biol Sex Differ

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Background The supraoptic nucleus (SON) of the hypothalamus contains magnocellular neurosecretory cells that secrete the hormones vasopressin and oxytocin. Sex differences in SON gene expression have been relatively unexplored. Our study used spatially resolved transcriptomics to visualize gene expression profiles in the SON of adult male (n = 4) and female (n = 4) Sprague-Dawley rats using Visium Spatial Gene Expression (10x Genomics). Methods Briefly, 10-μm coronal sections (~ 4 × 4 mm) containing the SON were collected from each rat and processed using Visium slides and recommended protocols. Data were analyzed using 10x Genomics’ Space Ranger and Loupe Browser applications and other bioinformatic tools. Two unique differential expression (DE) analysis methods, Loupe Browser and DESeq2, were used. Results Loupe Browser DE analysis of the SON identified 116 significant differentially expressed genes (DEGs) common to both sexes (e.g., Avp and Oxt), 31 significant DEGs unique to the males, and 73 significant DEGs unique to the females. DESeq2 analysis revealed 183 significant DEGs between the two groups. Gene Ontology (GO) enrichment and pathway analyses using significant genes identified via Loupe Browser revealed GO terms and pathways related to (1) neurohypophyseal hormone activity, regulation of peptide hormone secretion, and regulation of ion transport for the significant genes common to both males and females, (2) Gi signaling/G-protein mediated events for the significant genes unique to males, and (3) potassium ion transport/voltage-gated potassium channels for the significant genes unique to females, as some examples. GO/pathway analyses using significant genes identified via DESeq2 comparing female vs. male groups revealed GO terms/pathways related to ribosomal structure/function. Ingenuity Pathway Analysis (IPA) identified additional sex differences in canonical pathways (e.g., ‘Mitochondrial Dysfunction’, ‘Oxidative Phosphorylation’) and upstream regulators (e.g., CSF3, NFKB complex, TNF, GRIN3A). Conclusion There was little overlap in the IPA results for the two different DE methods. These results suggest sex differences in SON gene expression that are associated with cell signaling and ribosomal pathways.

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Section: Discussionsupporting

confidence: 76%

Section: Resultsmentioning

confidence: 67%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Spatial transcriptomics reveal basal sex differences in supraoptic nucleus gene expression of adult rats related to cell signaling and ribosomal pathways

Nguyen,

Duque,

Phillips

et al. 2023

Biol Sex Differ

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Isoflurane Rapidly Modifies Synaptic and Cytoskeletal Phosphoproteomes of the Supraoptic Nucleus of the Hypothalamus and the Cortex

Bárez-López,

Gadd,

Pauža

et al. 2023

Neuroendocrinology

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Introduction: Despite the widespread use of general anaesthetics, the mechanisms mediating their effects are still not understood. Although suppressed in most parts of the brain, neuronal activity, as measured by FOS activation, is increased in the hypothalamic supraoptic nucleus (SON) by numerous general anaesthetics, and evidence points to this brain region being involved in the induction of general anaesthesia and natural sleep. Posttranslational modifications of proteins, including changes in phosphorylation, enable fast modulation of protein function which could be underlying the rapid effects of general anaesthesia. In order to identify potential phosphorylation events in the brain mediating general anaesthesia effects, we have explored the phosphoproteome responses in the rat SON, and compared these to cingulate cortex (CC) which displays no FOS activation is response to general anaesthetics. Methods: Adult Sprague-Dawley rats were treated with isoflurane for 15 minutes. Proteins from the CC and SON were extracted and processed for Nano-LC Mass Spectrometry (LC-MS/MS). Phosphoproteomic determinations were performed by LC-MS/MS. Results: We found many changes in the phosphoproteomes of both the CC and SON in response to 15 minutes of isoflurane exposure. Pathway analysis indicated that proteins undergoing phosphorylation adaptations are involved in cytoskeleton remodelling and synaptic signalling events. Importantly, changes in protein phosphorylation appeared to be brain region-specific suggesting that differential phosphorylation adaptations might underlie the different neuronal activity responses to general anaesthesia between the CC and SON. Conclusion: In summary, these data suggest that rapid posttranslational modifications in proteins involved in cytoskeleton remodelling and synaptic signalling events might mediate the central mechanisms mediating general anaesthesia.

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Translational and Posttranslational Dynamics in a Model Peptidergic System

Cited by 2 publications

References 81 publications

Spatial transcriptomics reveal basal sex differences in supraoptic nucleus gene expression of adult rats related to cell signaling and ribosomal pathways

Spatial transcriptomics reveal basal sex differences in supraoptic nucleus gene expression of adult rats related to cell signaling and ribosomal pathways

Isoflurane Rapidly Modifies Synaptic and Cytoskeletal Phosphoproteomes of the Supraoptic Nucleus of the Hypothalamus and the Cortex

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