The Journal of Neuroscience, February 15, 1999, 19(4):1217-1225
Uptake of Precursor and Synthesis of Transmitter in a
Histaminergic Photoreceptor
Jennifer R.
Morgan1,
Kelley A.
Gebhardt2, and
Ann E.
Stuart2
1 Department of Neurobiology, Duke University Medical
Center, Durham, North Carolina 27710, and 2 Department of
Cell and Molecular Physiology, University of North Carolina at Chapel
Hill, Chapel Hill, North Carolina, 27599-7545
As a first step in understanding how the supply of the
neurotransmitter histamine is maintained in a photoreceptor, we
followed the uptake and metabolism of the immediate precursor of
histamine, histidine. [3H]Histidine taken
up into photoreceptors and glia was detected using autoradiography, and
synthesis of [3H]histamine from
[3H]histidine was assayed with thin-layer
chromatography. Photoreceptors from barnacles were pulsed (15 min) with
[3H]histidine (0.2-200 µM), then
maintained in normal saline for up to 24 hr. Autoradiography showed
that photoreceptor somata, axons, and presynaptic arbors were labeled,
but only weakly, like (nonhistaminergic) ganglion cells. Label instead
was concentrated over surrounding glia. Stimulating preparations with
light did not increase photoreceptor labeling. Grain counts from
photoreceptor axons showed uptake of [3H]histidine
into these neurons by a Na+-dependent mechanism with
a Km of ~50 µM. Over 24 hr
only 1% of the [3H]histidine taken up
by preparations was converted to [3H]histamine
either in the dark or in the light. Injections of [3H]histidine directly into photoreceptors
established that synthesis takes place within the photoreceptors and
confirmed that stimulation with light did not measurably affect the
rate of conversion of [3H]histidine to
[3H]histamine. These results suggest that
de novo synthesis of transmitter is unlikely to be as
important as its reuptake in maintaining neurotransmitter supply in
these photoreceptor terminals. In support of this conclusion,
photoreceptors accumulated more label when transmitter release was
stimulated with high K+ and histamine uptake was
antagonized with chlorpromazine.
Key words:
histamine; neurotransmitter synthesis; histidine uptake; photoreceptor; arthropod; barnacle
Copyright © 1999 Society for Neuroscience 0270-6474/99/1941217-09$05.00/0