A herpes simplex viral vector expressing green fluorescent protein can be used to visualize morphological changes in high-density neuronal culture

Torsten Falk
Department of Physiology
The University of Arizona
PO Box 245051
1609 N Warren Ave.
Tucson, AZ 85724, USA
Phone: 520-626-2044
Fax: 520-626-2383
E-mail: Tfalk@u.arizona.edu

Lori A. Strazdas
School of Public Health
Arizona Health Sciences Center
1501 N Campbell Ave.
Tucson, AZ 85724, USA
Phone: 520-626-6518
Fax: 520-626-6300
E-mail: Lstrazda@u.arizona.edu

Rebecca S. Borders
College of Medicine
Arizona Health Sciences Center
1501 N Campbell Ave.
Tucson, AZ 85724
Phone: 520-626-6518
Fax: 520-626-6300
E-mail: Bluntr@u.arizona.edu

Ramsey K. Kilani
College of Medicine
Arizona Health Sciences Center
1501 N Campbell Ave.
Tucson, AZ 85724, USA
Phone: 520-626-6518
Fax: 520-626-6300
E-mail: Rkk@u.arizona.edu

Andrea J. Yool
Department of Physiology
The University of Arizona
1609 N Warren Ave.
Tucson, AZ 85724, USA
Phone: 520-626-2198
Fax: 520-626-2383
E-mail: Ayool@u.arizona.edu

Scott J. Sherman*
Department of Neurology
College of Medicine
Arizona Health Sciences Center
PO Box 245023
1501 N Campbell Ave.
Tucson, AZ 85724-5023
Phone: 520-626-2319
Fax: 520-626-5999
E-mail: Ssherman@u.arizona.edu

*Corresponding author

Financial support: NIH grants 5 K08 NS 02015-03 (S.J.S.) and 1 R01 MH59747-01A1 (A.J.Y.); Epilepsy Foundation of America Fellowship (S.J.S.); and Small Grants Program funding from The University of Arizona (S.J.S.).

Keywords: cerebellum, green fluorescent protein, hippocampus, plasticity, Purkinje neuron.

High-density cultures of mammalian neurons offer a model system for studies of brain development, but the morphological features of individual neurons is difficult to ascertain. We show that a herpes virus vector expressing a bioluminescent protein allows detailed morphometric analyses of living neurons in complex culture environments. Expression of enhanced green fluorescent protein (eGFP) was constitutively driven in neurons using the herpes simplex virus amplicon system. This system allowed us to make novel observations regarding development in high-density cultures from rat hippocampus and cerebellum. After the phase of initial neurite outgrowth, maturing neurons continue to show rapid remodeling of the neurite branches (0.79 ± 0.11 mm/h per neurite; mean ± SEM, n=8), and displacement of the soma within the neurite arbor (1.35 ± 0.74 mm/h). These results demonstrate that a substantial capacity for morphological plasticity persists in maturing mammalian CNS neurons after cessation of net neurite outgrowth in early development.