In vivo AAV-mediated expression of calbindin-D28K in rat basal forebrain cholinergic neurons

Nicholas Nagykery*, Ernest F. Terwilliger, Changiz Geula

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

The cholinergic neurons of the basal forebrain (BFCNs) in human and non-human primates are rich in the calcium binding protein calbindin-D28k (CB). We have shown a selective loss of CB from BFCNs in the course of normal aging, which appears to predispose these neurons to tangle formation and degeneration in Alzheimer's disease. Our previous preliminary investigation demonstrated that rodent BFCNs are devoid of CB. Here we confirm that rat choline acetyltransferase-rich BFCNs are devoid of CB immunoreactivity. We then describe a method for adeno-associated viral vector (AAV) induced expression of CB in rat BFCNs in vivo. We constructed AAV vectors bearing the CB gene under the control of the CMV promoter, or neuron-specific enolase (NSE) promoter, to bias expression in neurons. Both vectors resulted in CB expression in mouse neuronal cultures, and in rat brain following injections. AAV-NSE-CB resulted in more robust expression in neurons. Injections of 10μl of AAV-NSE-CB in the BFCNs component located within the internal segment of globus pallidus and internal capsule resulted in expression of CB in 84% of BFCNs. Expression was optimum at 14 days. Injections of AAV-NSE-LacZ resulted in robust β-galactosidase expression, but no CB immunoreactivity. Our results show that use of NSE promoter leads to high expression of genes in neurons and that the BFCNs can be targeted for expression of genes that are differentially expressed in the rodent and primate brains. These findings have important implications for gene replacement therapy in human BFCNs.

Original languageEnglish (US)
Pages (from-to)106-113
Number of pages8
JournalJournal of Neuroscience Methods
Volume212
Issue number1
DOIs
StatePublished - 2013

Keywords

  • Adeno-associated viral vector
  • Aging
  • Alzheimer's disease
  • Basal forebrain cholinergic neurons
  • Calbindin-D

ASJC Scopus subject areas

  • Neuroscience(all)

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