Proteomic analysis identifies NPTX1 and HIP1R as potential targets of histone deacetylase-3-mediated neurodegeneration

Experimental Biology and Medicine
Zhe Qu, Santosh R D'Mello

Abstract

A defining feature of neurodegenerative diseases is the abnormal and excessive loss of neurons. One molecule that is particularly important in promoting neuronal death in a variety of cell culture and in vivo models of neurodegeneration is histone deacetylase-3 (HDAC3), a member of the histone deacetylase family of proteins. As a step towards understanding how HDAC3 promotes neuronal death, we conducted a proteomic screen aimed at identifying proteins that were regulated by HDAC3. HDAC3 was overexpressed in cultured rat cerebellar granule neurons (CGNs) and protein lysates were analyzed by mass spectrometry. Of over 3000 proteins identified in the screen, only 21 proteins displayed a significant alteration in expression. Of these, 12 proteins were downregulated whereas 9 proteins were upregulated. The altered expression of five of these proteins, TEX10, NPTX1, TFG, TSC1, and NFL, along with another protein that was downregulated in the proteomic screen, HIP1R, was confirmed using Western blots and commercially available antibodies. Because antibodies were not available for some of the proteins and since HDAC3 is a transcriptional regulator of gene expression, we conducted RT-PCR analysis to confirm expression changes. In separa...Continue Reading

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Citations

Jun 4, 2020·Experimental Biology and Medicine·Santosh R D'Mello
Jun 4, 2020·Particle and Fibre Toxicology·Sweelin ChewKatja M Kanninen
Feb 5, 2019·Genes & Genetic Systems·Jia LiJiajun Chen
Aug 17, 2021·Investigative Ophthalmology & Visual Science·Heather M SchmittRobert W Nickells
Aug 27, 2021·Frontiers in Molecular Neuroscience·Clara TejidoLudo Van Den Bosch

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Methods Mentioned

BETA
density gradient centrifugation
PCR
RNA-Seq
ChIP

Software Mentioned

GraphPad Prism
SINQ
IPA
Ingenuity
! Tandem
Open MS Search Algorithm ( OMSSA )

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