Matching neural morphology to molecular expression: single cell injection following immunostaining.
To match a neuron's morphology with its expression of a particular protein, it is useful to first identify the cell by immunostaining and then inject it with fluorescent dye. Such targeted injection cannot be performed with a hydrophilic dye (such as Lucifer yellow) because the neuron, once rendered porous to antibodies, does not retain it. But a lipophilic dye (such as DiI) injected iontophoretically into the soma forms a crystal and is thereby trapped. From this intracellular depot dye diffuses into the cell membrane to reveal the detailed morphology. We have used this strategy to identify the morphology of a GABAergic retinal bipolar cell and several types of GABAergic amacrine cell. In addition, we demonstrate probable connections from a narrow-field, GABAergic amacrine cell to the OFF brisk-transient ganglion cell. Finally, we show that the strategy works in the cortical slice, showing a layer IV cell immunostained for parvalbumin to be a "nest basket cell".
Kao
Yen-Hong
YH
Department of Neuroscience, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6058, USA. yen@mail.med.upenn.edu
Sterling
Peter
P
D025042:Amacrine Cells / Q000166:cytology / Q000378:metabolism; D000818:Animals; D002232:Carbocyanines; D002415:Cats; D048429:Cell Size / Q000502:physiology; D002540:Cerebral Cortex / Q000166:cytology / Q000378:metabolism; D004396:Coloring Agents; D003712:Dendrites / Q000378:metabolism / Q000648:ultrastructure; D005456:Fluorescent Dyes; D005968:Glutamate Decarboxylase / Q000378:metabolism; D006168:Guinea Pigs; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism; D008845:Microinjections / Q000379:methods*; D009433:Neural Inhibition / Q000502:physiology; D009434:Neural Pathways / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D009930:Organic Chemicals; D010320:Parvalbumins / Q000378:metabolism; D051381:Rats; D012160:Retina / Q000166:cytology* / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000378:metabolism; D009435:Synaptic Transmission / Q000502:physiology; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
C035005:3,3'-dihexadecylindocarbocyanine; D002232:Carbocyanines; D004396:Coloring Agents; D005456:Fluorescent Dyes; D007527:Isoenzymes; D009930:Organic Chemicals; D010320:Parvalbumins; C461159:SYTO 13; D005680:gamma-Aminobutyric Acid; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2