C. David Ross

Effects of trapezoid body and superior olive lesions on choline acetyltransferase activity in the rat cochlear nucleus.

Using a microdissection and quantitative microassay approach, choline acetyltransferase activities were mapped in the cochlear nuclei of rats having either transection of the trapezoid body or destruction of the superior olivary complex on one side in the brain stem. Lateral trapezoid body transection resulted in dramatic loss of choline acetyltransferase activity in all parts of the ipsilateral cochlear nucleus, while more medial transection had little effect. Destruction of the superior olivary complex resulted in dramatic loss of choline acetyltransferase activity in the ipsilateral cochlear nucleus, and detectable loss also contralaterally. The results suggest that most of the centrifugal cholinergic projections to the rat cochlear nucleus derive from or traverse the vicinity of the superior olivary complex bilaterally and enter the cochlear nucleus ventrally from the region of the trapezoid body.

Laminar Distributions of Choline Acetyltransferase and Acetylcholinesterase Activities in the Inner Plexiform Layer of Rat Retina

Abstract: Choline acetyltransferase and acetylcholinesterase activities were measured in samples taken at 7‐μm increments through the inner plexiform layer of rat retina. These enzyme activities were not uniformly distributed through the depth of the inner plexiform layer. Peaks of choline acetyltransferase activity occurred at about onethird and peaks of acetylcholinesterase activity at about one‐fifth of the depth into the inner plexiform layer from either side. The positions of the two peaks of choline acetyltransferase activity most likely correspond to the locations of processes from cholinergic amacrine somata in the inner nuclear layer, which spread in sublamina a, and processes from cholinergic amacrine somata “displaced’ in the ganglion cell layer which spread in sublamina b of the inner plexiform layer. The peaks of acetylcholinesterase activity may in addition correspond to the processes of cholinoceptive amacrine and ganglion cells. The magnitudes of choline acetyltransferase and acetylcholinesterase activities are as high as found anywhere in rat brain, emphasizing the important role of cholinergic mechanisms in visual processing through the rat inner plexiform layer.

Effect of olivocochlear bundle transection on choline acetyltransferase activity in the rat cochlear nucleus

Using a microdissection and quantitative microassay approach, choline acetyltransferase activities were mapped in the cochlear nuclei of rats having complete transections of the olivocochlear bundle on one side in the brain stem. In rats in which the trapezoid body was not significantly damaged by the lesion, consistent reductions of choline acetyltransferase activity in subregions of the lesion-side cochlear nucleus, as compared to the control side, averaged about 20%. Nevertheless, a profound lesion-side reduction of choline acetyltransferase activity was found in a branch connection from the olivocochlear bundle to the cochlear nucleus. The results suggest that branches from the olivocochlear bundle are cholinergic, but contribute a relatively minor proportion of the cholinergic synapses in all regions of the rat cochlear nucleus. In the light of previous results with more extensive lesions, it can be proposed that synapses in all regions of the rat cochlear nucleus. In the light of previous results with more extensive lesions, it can be proposed that most cholinergic input into the rat cochlear nucleus enters by a ventral route along the trapezoid body. It is noted that this represents a quantitatively somewhat different situation from that in the cat.

Effects of large brain stem lesions on the cholinergic system in the rat cochlear nucleus

Large lesions were made medial to one cochlear nucleus in rats, in order to cut virtually centrifugal pathways to it. To estimate the contribution of these centrifugal pathways to cholinergic synapses in the cochlear nucleus, choline acetyltransferase and acetylcholinesterase activities were mapped, by quantitative histochemical procedures, in lesion and control side cochlear nuclei. Choline acetyltransferase activities were reduced by 85-90% in most regions of the lesion side cochlear nucleus and by 65-75% in granular regions. Acetylcholinesterase activities were reduced by 50% or less in the same regions. The choline acetyltransferase results are consistent with a conclusion that by far most cholinergic synapses in the rat cochlear nucleus derive from centrifugal pathways. Additionally, the effects of the lesions on enzyme activities in the lateral superior olivary nucleus and ventral nucleus of the trapezoid body, and in the facial, motor trigeminal, and spinal trigeminal nuclei were examined. In the lesion side facial nucleus, 60% and 40% decreases in choline acetyltransferase and acetylcholinesterase activities, respectively, were apparently consequences of facial root transection. Lesion-control enzyme activity differences in the other nuclei were much smaller.

Contribution of centrifugal innervation to choline acetyltransferase activity in the cat cochlear nucleus

Using a quantitative microchemical mapping approach combined with surgical cuts of fiber tracts, the contributions of centrifugal pathways to choline acetyltransferase activity were mapped three-dimensionally in the cat cochlear nucleus. Large reductions of choline acetyltransferase activity, averaging 70%, were measured in almost all parts of the lesion-side nucleus following transection of virtually all its centrifugal connections. More superficial cuts, penetrating just through the olivocochlear bundle, also led to significant reductions of enzyme activity, especially most rostrally in the anteroventral cochlear nucleus and superficial granular region, where the reductions were similar to those following the complete cuts. Lesions encroaching upon the superior olivary complex gave bilateral effects. Transverse cuts between rostral and caudal parts of the cochlear nucleus gave some small effects. The results suggest that, as in rats, most choline acetyltransferase activity in the cat cochlear nucleus is associated with its centrifugal innervation. However, unlike the situation in rats, the enzyme activity in cats is related more to olivocochlear branches than to ventral fibers in the trapezoid body region. Also, the choline acetyltransferase activity related to olivocochlear collateral innervation is much less uniformly distributed within the cochlear nucleus in cats than in rats.

Aspartate Aminotransferase Activity in Fiber Tracts of the Rat Brain

Abstract: Activity of aspartate aminotransferase, an enzyme which catalyzes the interconversion of the excitatory transmitter candidates, glutamate and aspartate, has been measured in fiber tracts of rat, with an emphasis on sensory and motor systems of the brain. Most tracts had significantly higher activities than the cholinergic facial nerve root, consistent with the possibility that a component of aspartate aminotransferase activity might serve as a marker for neurons using glutamate and/or aspartate as neurotransmitter. Highest activity was in the auditory nerve root. On the other hand, a close correlation was found between aspartate aminotransferase and malate de‐hydrogenase activities in the fiber tracts, raising the question whether aspartate aminotransferase activity may be more closely related to energy metabolism than to transmitter metabolism.

Quantitative distributions of aspartate aminotransferase and glutaminase activities in the rat cochlea

The intra-cochlear distributions of aspartate aminotransferase and glutaminase, prominent enzymes of aspartate and glutamate metabolism, have been studied by quantitative microchemical techniques. Also measured was choline acetyltransferase, the enzyme synthesizing acetylcholine, and a marker for the olivocochlear bundle. Aspartate aminotransferase activity was highest in the stria vascularis, about half this high in the organ of Corti synaptic (hair cell) zones, somewhat lower in the organ of Corti non-synaptic (Hensens cell) zones, lower yet in Reissners and lowest in the tectorial membrane. Glutaminase, on the other hand, had its highest activity in synaptic zones, about a third of that activity in the organ of Corti non-synaptic zones, and a barely detectable activity in Reissners and tectorial membranes, and stria vascularis. Seven days after transection of the olivocochlear bundle, no significant difference was found between lesion- and control-side aspartate aminotransferase or glutaminase activities, even though no choline acetyltransferase activity remained in the lesion-side of the organ of Corti. Both the distribution of aspartate aminotransferase activity and the lesion results would seem to implicate it in energy more so than neurotransmitter metabolism. The distribution of glutaminase activity could be consistent with a role in neurotransmission; however, the lesion data were unable to demonstrate a specific association with the olivocochlear bundle.

Quantitative distribution of six amino acids in rat retinal layers

Concentrations of glutamate, aspartate, glutamine, glycine, GABA, and taurine were determined in samples microdissected from rat retinal layers and assayed by HPLC. Glutamate and glutamine were relatively high in the inner nuclear (INL) and ganglion cell (GCL) layers; aspartate was relatively high in the outer nuclear layer (ONL), outer plexiform layer, and INL. Distributions of glutamate and aspartate did not correlate well with those of enzymes involved in their metabolism. Glycine and GABA were highest in the inner plexiform layer, with increasing concentrations through the INL, and were relatively high in the GCL. Taurine was highest in the ONL.

Effect of enucleation on choline acetyltransferase activity in layers of goldfish optic tectum

Choline acetyltransferase (ChAT) activity was determined in layers of optic tectum in control goldfish and in goldfish 4-20 days following unilateral enucleation. Significant changes in activity were found in the periventricular (PV) and superficial gray and white (SGW) layers. Within 4 days, ChAT activity in the PV layer on the lesioned side was about 75% of that on the control side. By 20 days, ChAT specific activity in the SGW layer on the lesioned side was about 150-160% of that on the control side. This increase in specific activity in the SGW layer was accounted for by the decrease in volume and in density of the layer after enucleation, so that the total amount of activity in the layer did not change significantly, indicating that the optic terminals contain little to no ChAT activity. ChAT activity in the optic tract was very low and did not decrease after enucleation. These data strongly indicate that the retinotectal pathway in goldfish is not cholinergic and, therefore, that the ChAT activity in the SGW layer is related to sources other than retinal ganglion cells. It is suggested that one such source might be neurons with somata in the PV layer.

Tumor grade, microvessel density, and activities of malate dehydrogenase, lactate dehydrogenase, and hexokinase in squamous cell carcinoma.

Squamous cell carcinomas were evaluated with respect to tumor differentiation (through use of hematoxylin and eosin stain), microvessel density (through use of CD-34 immunocytochemical stain), and magnitudes of malate dehydrogenase (MDH), hexokinase, and lactate dehydrogenase (LDH) enzyme activities. Direct correlations were found between tumor grade, MDH activity, and microves-sel density. Direct correlations were also found between hexokinase activity and MDH activity and microvessel density. Inverse correlations were found between LDH activity and both tumor grade and MDH activity. These results suggest that the high rate of glucose utilization (indicated by hexokinase activity) found in more poorly differentiated tumors has a higher component of aerobic oxidative metabolism (indicated by MDH activity) and a relatively lower contribution from anaerobic metabolism (indicated by LDH activity) than do the rates found in more differentiated tumors. It is also suggested that as the glycolytic rate increases, more pyruvate goes into the Krebs cycle than into lactate. The availability of glucose-derived pyruvate for oxidative metabolism would mean less of a dependency on glutamine as a carbon source in squamous cell carcinoma.