Awardee OrganizationUNIVERSITY OF ALABAMA AT BIRMINGHAM
Description
Abstract Text
This study is directed at elucidating the morphological and physiological
organization of the inner plexiform layer of the mammalian retina. It
focuses on examining the ultrastructure of physiologically identified
ganglion cells in the rabbit retina, and will help interpret our existing
morphological data pertaining to the synaptic profiles of cone bipolar
axonal terminals with amacrine and ganglion cells. Intracellular
recordings are obtained under visual observations using infrared
illumination and DIC optics on an isolated, superfused retina preparation.
These recordings will determine: whether the cells respond to the onset or
termination of light in a sustained or transient manner; the nature of
their photoreceptor input; and whether the receptive field properties
undergo any reorganization with changes in the adaptive state of the
cells. Once physiologically identified, the cells are injected with
Lucifer Yellow to verify their morphology with the light microscope; the
Lucifer is photoconverted to an osmophilic precipitate, which can be
visualized with the electron microscope; and serial micrographs of the
labeled cell are used in computer graphics to reconstruct the synaptic
connections of several major dendrites. Special attention will be focused
on ascertaining a relationship between the amount of amacrine and cone
bipolar cell input the ganglion cell receives with respect to its
physiological responses to light. A new technique has been developed for
the simultaneous, ultrastructural examination of two different
morphologically and physiologically identified neurons to demonstrate that
they form the pre- and postsynaptic processes of a synapse. This method
employs the use of separate intracellular injections of HRP and a
fluorescent dye (Lucifer Yellow) into different cells which have
overlapping dendritic trees and are suspected of synapsing with each
other. The tissue is sequentially reacted under two different conditions
using two different chromogens: henzidine dihydrochloride is used to react
the HRP, and diaminobenzidine is used to form the reaction product during
the photoconversion of Lucifer. The fluorescent dye cannot be
photoconverted in the presence of any chromogen unless it is maximally
excited with its specific wavelength light. These two chromogens produce
distinctly different appearing reaction products that can be
differentiated at the light and electron microscope. This method will be
used to determine whether the DAPI-accumulating, flat cone bipolar cells
synapse with off-ganglion cells in sublamina alpha of the IPL, and if the
starburst amacrine cells synapse with the on-off directionally selective
(DS) ganglion cells. Variations in the synaptic profiles among amacrine,
bipolar and DS ganglion cells, for which the preferred-null axis has been
identified, will be examined to evaluate the morphological substrate for
the physiological models of DS ganglion cells. Finally, by obtaining
intracellular recordings from DAPI-accumulating cone bipolar cells, and
injecting them with Lucifer to correlate their morphology with the
previously examined cells, this study will attempt to provide conclusive
evidence that the are off-bipolar cells.
No Sub Projects information available for 5R01EY003011-20
Publications
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