Cyan fluorescent protein expression in ganglion and amacrine cells in a thy1-CFP transgenic mouse retina.
OBJECTIVE
To characterize cyan fluorescent protein (CFP) expression in the retina of the thy1-CFP (B6.Cg-Tg(Thy1-CFP)23Jrs/J) transgenic mouse line.


METHODS
CFP expression was characterized using morphometric methods and immunohistochemistry with antibodies to neurofilament light (NF-L), neuronal nuclei (NeuN), POU-domain protein (Brn3a) and calretinin, which immunolabel ganglion cells, and syntaxin 1 (HPC-1), glutamate decarboxylase 67 (GAD(67)), GABA plasma membrane transporter-1 (GAT-1), and choline acetyltransferase (ChAT), which immunolabel amacrine cells.


RESULTS
CFP was extensively expressed in the inner retina, primarily in the inner plexiform layer (IPL), ganglion cell layer (GCL), nerve fiber layer, and optic nerve. CFP fluorescent cell bodies were in all retinal regions and their processes ramified in all laminae of the IPL. Some small, weakly CFP fluorescent somata were in the inner nuclear layer (INL). CFP-containing somata in the GCL ranged from 6 to 20 microm in diameter, and they had a density of 2636+/-347 cells/mm2 at 1.5 mm from the optic nerve head. Immunohistochemical studies demonstrated colocalization of CFP with the ganglion cell markers NF-L, NeuN, Brn3a, and calretinin. Immunohistochemistry with antibodies to HPC-1, GAD(67), GAT-1, and ChAT indicated that the small, weakly fluorescent CFP cells in the INL and GCL were cholinergic amacrine cells.


CONCLUSIONS
The total number and density of CFP-fluorescent cells in the GCL were within the range of previous estimates of the total number of ganglion cells in the C57BL/6J line. Together these findings suggest that most ganglion cells in the thy1-CFP mouse line 23 express CFP. In conclusion, the thy1-CFP mouse line is highly useful for studies requiring the identification of ganglion cells.
Raymond
Iona D
ID
Department of Neurobiology, David Geffen School of Medicine at University of California Los Angeles, Los Angeles, CA 90095-1763, USA. iona.d.raymond@ucla.edu
Vila
Alejandro
A
Huynh
Uyen-Chi N
UC
Brecha
Nicholas C
NC
D025042:Amacrine Cells / Q000378:metabolism*; D000818:Animals; D004268:DNA-Binding Proteins; D050485:GABA Plasma Membrane Transport Proteins / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D009900:Optic Nerve / Q000166:cytology / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000378:metabolism*; D050827:Syntaxin 1 / Q000378:metabolism; D018800:Thy-1 Antigens / Q000378:metabolism*
C485182:Cyan Fluorescent Protein; D004268:DNA-Binding Proteins; D050485:GABA Plasma Membrane Transport Proteins; D009419:Nerve Tissue Proteins; C492380:NeuN protein, mouse; D009687:Nuclear Proteins; C494214:Stx1a protein, mouse; D050827:Syntaxin 1; D018800:Thy-1 Antigens; D049452:Green Fluorescent Proteins