Expression analysis of green fluorescent protein in retinal neurons of four transgenic mouse lines

Neuroscience. 2009 Apr 21;160(1):126-39. doi: 10.1016/j.neuroscience.2009.01.081. Epub 2009 Feb 13.

Abstract

Transgenic mice that express enhanced green fluorescent protein (EGFP) under the control of a cell-specific promoter have been used with great success to identify and label specific cell types of the retina. We studied the expression of EGFP in the retina of mice making use of four transgenic mouse lines. Expression of EGFP driven by the calretinin promoter was found in amacrine, displaced amacrine and ganglion cells. Comparison of the EGFP expression and calretinin immunolabeling showed that many but not all cells appear to be double labeled. Expression of EGFP under the control of the choline acetyltransferase promoter was found in amacrine cells; however, the cells did not correspond to the well known cholinergic (starburst) cells of the mouse retina. The expression of EGFP under the control of the parvalbumin promoter was restricted to amacrine cells of the inner nuclear layer and to cells of the ganglion cell layer (displaced amacrine cells and ganglion cells). Most of the cells were also immunoreactive for parvalbumin, however, differences in labeling intensity were observed. The expression of EGFP driven by the promoter for the 5-HT3 A receptor (5-HTR3A) was restricted to type 5 bipolar cells. In contrast, immunostaining for 5-HTR3A was found in synaptic hot spots in sublamina 1 of the inner plexiform layer and was not related to type 5 bipolar cells. The results show that these transgenic mice are very useful for future electrophysiological studies of specific types of amacrine and bipolar cells that express EGFP and thus permit directed microelectrode targeting under microscopic control.

MeSH terms

  • Acetylcholine / metabolism
  • Amacrine Cells / metabolism
  • Animals
  • Calbindin 2
  • Choline O-Acetyltransferase / genetics
  • Gene Expression*
  • Green Fluorescent Proteins / genetics*
  • Green Fluorescent Proteins / metabolism*
  • Immunohistochemistry
  • Mice
  • Mice, Transgenic
  • Microscopy, Fluorescence
  • Parvalbumins / genetics
  • Promoter Regions, Genetic
  • Receptors, Serotonin, 5-HT3 / genetics
  • Retinal Bipolar Cells / metabolism
  • Retinal Ganglion Cells / metabolism
  • Retinal Neurons / metabolism*
  • S100 Calcium Binding Protein G / genetics
  • Species Specificity

Substances

  • Calb2 protein, mouse
  • Calbindin 2
  • Parvalbumins
  • Receptors, Serotonin, 5-HT3
  • S100 Calcium Binding Protein G
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Choline O-Acetyltransferase
  • Acetylcholine