Expression Profiling of RNA Transcripts during Neuronal Maturation and Ischemic Injury

Abstract

Neuronal development is a pro-survival process that involves neurite growth, synaptogenesis, synaptic and neuronal pruning. During development, these processes can be controlled by temporal gene expression that is orchestrated by both long non-coding RNAs and microRNAs. To examine the interplay between these different components of the transcriptome during neuronal differentiation, we carried out mRNA, long non-coding RNA and microRNA expression profiling on maturing primary neurons. Subsequent gene ontology analysis revealed regulation of axonogenesis and dendritogenesis processes by these differentially expressed mRNAs and non-coding RNAs. Temporally regulated mRNAs and their associated long non-coding RNAs were significantly over-represented in proliferation and differentiation associated signalling, cell adhesion and neurotrophin signalling pathways. Verification of expression of the Axin2, Prkcb, Cntn1, Ncam1, Negr1, Nrxn1 and Sh2b3 mRNAs and their respective long non-coding RNAs in an in vitro model of ischemic-reperfusion injury showed an inverse expression profile to the maturation process, thus suggesting their role(s) in maintaining neuronal structure and function. Furthermore, we propose that expression of the cell adhesion molecules, Ncam1 and Negr1 might be tightly regulated by both long non-coding RNAs and microRNAs.

MicroRNAs in Hyperglycemia Induced Endothelial Cell Dysfunction

“…We found that the expression of miR-140-5p to be increased in IFG as well as in T2DM, in both the human and rat miRNA microarray analyses, as well as in our in vitro study, where miR-140-5p level increased with increasing glucose concentrations ( Table 1 and Table S2 [ 72 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 , 106 , 107 , 108 , 109 , 110 , 111 , 112 , 113 , 114 , 115 , 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , 124 , 125 , 126 , 127 , 128 , 129 , 130 , 131 , 132 , 133 , 134 , 135 ]). This increase was also found to correlate with the increase in apoptosis ( Figure 2 ).…”

Hyperactive somatostatin interneurons contribute to excitotoxicity in neurodegenerative disorders

“…Primary antibodies used were mouse anti-parvalbumin 25 (1:1000 dilution, p3088, Sigma-Aldrich, St. Louis, MO), rabbit anti-VGAT 21 (1:2000 dilution, 131003, Synaptic Systems, Germany), rabbit anti-somatostatin 26 (1:300 dilution, sc-13099, Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-NeuN 27 (1:1000 dilution, ab104224, Abcam, Cambridge, MA) and mouse anti-ubiquitin 28 (1:500 dilution, 3936, Cell Signaling Technology, Beverly, MA). Secondary antibodies used were Alexa Fluor 647 donkey anti-mouse IgG 29 (1:300 dilution, 715-605-150, Jackson ImmunoResearch Laboratories, West Grove, PA) and Alexa Fluor 647 donkey anti-rabbit IgG 30 (1:300 dilution, 711-605-152, Jackson ImmunoResearch Laboratories).…”

MiR-335 Regulates Hif-1α to Reduce Cell Death in Both Mouse Cell Line and Rat Ischemic Models

“…Primary cultures of cortical neurons were established from E15 Swiss albino mouse brains according to Hirai et al [ 28 ] with slight modifications performed by Kaur et al [ 29 ]. The cortices were dissected from E15 mouse embryos and washed with Hanks’ balanced salt solution (HBSS; Gibco, Invitrogen, USA).…”