Inflammation-induced reversible switch of the neuron-specific enolase promoter from Purkinje neurons to Bergmann glia

Abstract

Neuron-specific enolase (NSE) is a glycolytic isoenzyme found in mature neurons and cells of neuronal origin. Injecting adeno-associated virus serotype 9 (AAV9) vectors carrying the NSE promoter into the cerebellar cortex is likely to cause the specific transduction of neuronal cells, such as Purkinje cells (PCs) and interneurons, but not Bergmann glia (BG). However, we found BG-predominant transduction without PC transduction along a traumatic needle tract for viral injection. The enhancement of neuroinflammation by the co-application of lipopolysaccharide (LPS) with AAV9 significantly expanded the BG-predominant area concurrently with the potentiated microglial activation. The BG-predominant transduction was gradually replaced by the PC-predominant transduction as the neuroinflammation dissipated. Experiments using glioma cell cultures revealed significant activation of the NSE promoter due to glucose deprivation, suggesting that intracellularly stored glycogen is metabolized through the glycolytic pathway for energy. Activation of the glycolytic enzyme promoter in BG concurrently with inactivation in PC may have pathophysiological significance for the production of lactate in activated BG and the utilization of lactate, which is provided by the BG-PC lactate shuttle, as a primary energy resource in injured PCs.

Minimal Purkinje Cell-Specific PCP2/L7 Promoter Virally Available for Rodents and Non-human Primates

“…Since the identification of the L7-4 sequence as an effective Purkinje cell-specific promoter suitable for viral vectors, the promoter has been widely used to date. 28 , 29 , 30 , 31 Nevertheless, we and others have noted a diffuse transduction in non-Purkinje cells in the molecular and granule cell layers. In addition, the L7 splice variants that use different ATG in exon 1 as a translation initiation codon (forms B and C) prompted us to narrow down the L7 further to a shorter promoter with a more restricted leakage to other cell types.…”

“…In a yet unpublished previous study using lentiviral vectors, we found that a 1-kb fragment of the L7 promoter exhibited strikingly greater promoter strength than the original 3-kb L7 promoter (US Patent 8912315 B2), 27 although the experimental procedure to obtain the 1-kb fragment and the sequence have not yet been published. Subsequent utilization of the compact 1-kb promoter by several groups, including ours, 28 , 29 , 30 , 31 revealed overall maintenance of the Purkinje cell selectivity, but with some diffuse activity in off-target cells.…”

Lactate Shuttles in Neuroenergetics—Homeostasis, Allostasis and Beyond

“…The concept of lactate shuttle systems extends to other types of neuronal cells, such as the oligodendrocyte–axon lactate shuttle (Baltan, 2015 ) and the Bergmann glia (BG) Purkinje cell (PC) lactate shuttle (Sawada et al, 2016 ). Lactate shuttles extend beyond the brain—they are of particular interest in cancer research (Pizzuto et al, 2012 ).…”

Synergistic effects of Rho kinase inhibitor Y‐27632 and Noggin overexpression on the proliferation and neuron‐like cell differentiation of stem cells derived from human exfoliated deciduous teeth

“…RT‐qPCR and Western blot assay analyses indicated that in comparison to the cells in Noggin group, the cells in Noggin + Y‐27632 group showed an significantly elevated expression of NES, Nestin, and GFAP. NSE, a glycolytic isoenzyme, is found in mature neurons and cells of neuronal origin . Binding of overexpressed Noggin to NSE promoter (NSE‐Noggin) has been shown to enhance neurological functions like learning and memory, which are associated with BMP signaling components in the hippocampus .…”

“…NSE, a glycolytic isoenzyme, is found in mature neurons and cells of neuronal origin. 27 Binding of overexpressed Noggin to NSE promoter (NSE-Noggin) has been shown to enhance neurological functions like learning and memory, which are associated with BMP signaling components in the hippocampus. 28 Moreover, in agreement to our results, a previous study has also proved that bone marrow derived mesenchymal stem cells transfected with Noggin gene can differentiate into neuron-like cells in vitro, and such activity is associated with the expression of NSE, neurofilament 200, neuronal nuclei, and GFAP.…”