Investigating Methodological Differences in the Assessment of Dendritic Morphology of Basolateral Amygdala Principal Neurons-A Comparison of Golgi-Cox and Neurobiotin Electroporation Techniques

Paul M Klenowski; Sophie E Wright; Erica W H Mu; Peter G Noakes; Nickolas A Lavidis; Selena E Bartlett; Mark C Bellingham; Matthew J Fogarty

doi:10.3390/brainsci7120165

Investigating Methodological Differences in the Assessment of Dendritic Morphology of Basolateral Amygdala Principal Neurons-A Comparison of Golgi-Cox and Neurobiotin Electroporation Techniques

Brain Sci. 2017 Dec 19;7(12):165. doi: 10.3390/brainsci7120165.

Authors

Paul M Klenowski^{1

2}, Sophie E Wright³, Erica W H Mu⁴, Peter G Noakes^{5

6}, Nickolas A Lavidis⁷, Selena E Bartlett⁸, Mark C Bellingham⁹, Matthew J Fogarty^{10

11}

Affiliations

¹ Translational Research Institute, Queensland University of Technology, Brisbane 4102, Australia. paul.klenowski@umassmed.edu.
² Department of Neurobiology, University of Massachusetts Medical School, Worcester, MA 01605, USA. paul.klenowski@umassmed.edu.
³ School of Biomedical Sciences, The University of Queensland, Brisbane 4072, Australia. sophiewright@live.com.au.
⁴ School of Biomedical Sciences, The University of Queensland, Brisbane 4072, Australia. e.mu@uq.edu.au.
⁵ School of Biomedical Sciences, The University of Queensland, Brisbane 4072, Australia. p.noakes@uq.edu.au.
⁶ Queensland Brain Institute, the University of Queensland, Brisbane 4072, Australia. p.noakes@uq.edu.au.
⁷ School of Biomedical Sciences, The University of Queensland, Brisbane 4072, Australia. lavidis@uq.edu.au.
⁸ Translational Research Institute, Queensland University of Technology, Brisbane 4102, Australia. selena.bartlett@qut.edu.au.
⁹ School of Biomedical Sciences, The University of Queensland, Brisbane 4072, Australia. mark.bellingham@uq.edu.au.
¹⁰ School of Biomedical Sciences, The University of Queensland, Brisbane 4072, Australia. m.fogarty@uq.edu.au.
¹¹ Department of Physiology and Biomedical Engineering, Mayo Clinic College of Medicine, Rochester, MN 55905, USA. m.fogarty@uq.edu.au.

Abstract

Quantitative assessments of neuronal subtypes in numerous brain regions show large variations in dendritic arbor size. A critical experimental factor is the method used to visualize neurons. We chose to investigate quantitative differences in basolateral amygdala (BLA) principal neuron morphology using two of the most common visualization methods: Golgi-Cox staining and neurobiotin (NB) filling. We show in 8-week-old Wistar rats that NB-filling reveals significantly larger dendritic arbors and different spine densities, compared to Golgi-Cox-stained BLA neurons. Our results demonstrate important differences and provide methodological insights into quantitative disparities of BLA principal neuron morphology reported in the literature.

Keywords: Golgi–Cox; basolateral amygdala; dendrites; neurobiotin; principal neuron; spines.