Journal article

Using a Multiplex Nucleic Acid in situ Hybridization Technique to Determine HCN4 mRNA Expression in the Adult Rodent Brain

Julia Oyrer, Lauren E Bleakley, Kay L Richards, Snezana Maljevic, A Marie Phillips, Steven Petrou, Cameron J Nowell, Christopher A Reid

FRONTIERS IN MOLECULAR NEUROSCIENCE | FRONTIERS MEDIA SA | Published : 2019

Abstract

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels carry a non-selective cationic conductance, I h , which is important for modulating neuron excitability. Four genes (HCN1-4) encode HCN channels, with each gene having distinct expression and biophysical profiles. Here we use multiplex nucleic acid in situ hybridization to determine HCN4 mRNA expression within the adult mouse brain. We take advantage of this approach to detect HCN4 mRNA simultaneously with either HCN1 or HCN2 mRNA and markers of excitatory (VGlut-positive) and inhibitory (VGat-positive) neurons, which was not previously reported. We have developed a Fiji-based analysis code that enables quantification of mRNA..

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Grants

Awarded by National Health and Medical Research Council (NHMRC)


Awarded by NHMRC


Funding Acknowledgements

This work was supported by the National Health and Medical Research Council (NHMRC) Program Grant (10915693) to SP and CR and an NHMRC Project Grant (1143101) to CR. LB acknowledges the support of an Australian Government Research Training Program Scholarship.