Reference Genes across Nine Brain Areas of Wild Type and Prader-Willi Syndrome Mice: Assessing Differences in Igfbp7, Pcsk1, Nhlh2 and Nlgn3 Expression

Kummerfeld, Delf-Magnus; Skryabin, Boris V.; Brosius, Juergen; Vakhrushev, Sergey Y.; Rozhdestvensky, Timofey S.

doi:10.3390/ijms23158729

Cited by 2 publications

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“…The latter include validation of (B) 10 down-regulated genes ( Derl3 , Mylip , Atp10a , Jph3 , Ndrg4 , Tap1 , Tap2 , Syt1 , Robo2 , Id4 ), as well as (C) for a set of 4 upregulated DEGs involved in the secretory pathway ( Chgb , Cacna1a , Tmem176a , Tmem176b ), with apparent upregulation for an exogenous human insulin ( Hs , Homo sapiens )-neomycin resistance ( Neo R) transgene in the INS-1 lines, although the latter is artifactual due to transgene silencing in line 16 (see S16A , S16I–S16K and S16M Fig ). (D,E) However, the RT-ddPCR analyses failed to validate (D) several candidate upregulated DEGs from the RNA-seq data ( Fig 4A ), or (E) candidates for downregulated DEGs based on data from PWS iPSC-derived neuronal and Snord116 -deficient mouse models [ 60 , 129 ]; likewise, another mouse study [ 130 ] did not confirm Pcsk1 . Statistical comparison by Welch’s t-test: *, P < 0.05; **, P < 0.005; ***, P < 0.0005.…”

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confidence: 99%

Insulin secretion deficits in a Prader-Willi syndrome β-cell model are associated with a concerted downregulation of multiple endoplasmic reticulum chaperones

et al. 2023

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Prader-Willi syndrome (PWS) is a multisystem disorder with neurobehavioral, metabolic, and hormonal phenotypes, caused by loss of expression of a paternally-expressed imprinted gene cluster. Prior evidence from a PWS mouse model identified abnormal pancreatic islet development with retention of aged insulin and deficient insulin secretion. To determine the collective roles of PWS genes in β-cell biology, we used genome-editing to generate isogenic, clonal INS-1 insulinoma lines having 3.16 Mb deletions of the silent, maternal- (control) and active, paternal-allele (PWS). PWS β-cells demonstrated a significant cell autonomous reduction in basal and glucose-stimulated insulin secretion. Further, proteomic analyses revealed reduced levels of cellular and secreted hormones, including all insulin peptides and amylin, concomitant with reduction of at least ten endoplasmic reticulum (ER) chaperones, including GRP78 and GRP94. Critically, differentially expressed genes identified by whole transcriptome studies included reductions in levels of mRNAs encoding these secreted peptides and the group of ER chaperones. In contrast to the dosage compensation previously seen for ER chaperones in Grp78 or Grp94 gene knockouts or knockdown, compensation is precluded by the stress-independent deficiency of ER chaperones in PWS β-cells. Consistent with reduced ER chaperones levels, PWS INS-1 β-cells are more sensitive to ER stress, leading to earlier activation of all three arms of the unfolded protein response. Combined, the findings suggest that a chronic shortage of ER chaperones in PWS β-cells leads to a deficiency of protein folding and/or delay in ER transit of insulin and other cargo. In summary, our results illuminate the pathophysiological basis of pancreatic β-cell hormone deficits in PWS, with evolutionary implications for the multigenic PWS-domain, and indicate that PWS-imprinted genes coordinate concerted regulation of ER chaperone biosynthesis and β-cell secretory pathway function.

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Section: Supporting Informationmentioning

confidence: 99%