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Published on: April 29, 2015
Distribution of high affinity choline transporter immunoreactivity in the primate central nervous system
Laura Kus1, Ewa Borys, Ya Ping Chu
1Department of Neurological Sciences, Rush Presbyterian-St. Luke's Medical Center, Chicago, Illinois 60612.
Insights
This study maps the high-affinity choline transporter (CHT) in the primate central nervous system, identifying its presence in key cholinergic neurons and fibers across the brain and spinal cord. The findings highlight CHT as a valuable marker for cholinergic function in aging and neurological diseases.
Area of Science:
- Neuroscience
- Neuroanatomy
- Cholinergic System Research
Background:
- The high-affinity choline transporter (CHT) is crucial for synthesizing the neurotransmitter acetylcholine.
- Understanding CHT distribution is vital for investigating cholinergic neuronal function and related disorders.
- Previous studies have primarily focused on rodent models, necessitating primate-specific data.
Purpose of the Study:
- To determine the distribution of CHT-immunoreactive profiles in the central nervous system (CNS) of the monkey.
- To validate a novel monoclonal antibody (clone 62-2E8) for labeling CHT in primate CNS.
- To investigate the co-localization of CHT with choline acetyltransferase (ChAT) in primate neurons.
Main Methods:
- Immunohistochemistry using a mouse monoclonal antibody against human recombinant CHT.
- Analysis of CHT distribution in monkey telencephalon, brainstem, and spinal cord.
- Dual fluorescent confocal microscopy for co-localization studies with ChAT.
Main Results:
- CHT immunoreactivity was observed in perikarya and fibers across various monkey CNS regions, including the striatum, hippocampus, amygdala, and brainstem nuclei.
- A similar distribution pattern of CHT was found in the aged human brain and spinal cord.
- The majority of CHT-immunoreactive neurons co-localized with the cholinergic marker choline acetyltransferase (ChAT).
Conclusions:
- The validated CHT antibody effectively labels cholinergic structures in the primate CNS.
- This study provides a detailed map of CHT distribution in the primate brain and spinal cord.
- CHT serves as a significant marker for studying cholinergic neuronal function, aging, and neurological diseases in primates.
Abstract:
A mouse monoclonal antibody (clone 62-2E8) raised against a human recombinant high-affinity choline transporter (CHT)-glutathione-S-transferase fusion protein was used to determine the distribution of immunoreactive profiles containing this protein in the monkey central nervous system (CNS). Within the monkey telencephalon, CHT-immunoreactive perikarya were found in the striatum, nucleus accumbens, medial septum, vertical and horizontal limb nuclei of the diagonal band, nucleus basalis complex, and the bed nucleus of the stria terminalis. Dense fiber staining was observed within the islands of Calleja, olfactory tubercle, hippocampal complex, amygdala; moderate to light fiber staining was seen in iso- and limbic cortices. CHT-containing fibers were also present in sensory and limbic thalamic nuclei, preoptic and hypothalamic areas, and the floccular lobe of the cerebellum. In the brainstem, CHT-immunoreactive profiles were observed in the pedunculopontine and dorsolateral tegmental nuclei, the Edinger-Westphal, oculomotor, trochlear, trigeminal, abducens, facial, ambiguus, dorsal vagal motor, and hypoglossal nuclei. In the spinal cord, CHT-immunoreactive ventral horn motoneurons were seen in close apposition to intensely immunoreactive C-terminals at the level of the cervical spinal cord. CHT immunostaining revealed a similar distribution of labeled profiles in the aged human brain and spinal cord. Dual fluorescent confocal microscopy revealed that the majority of CHT immunoreactive neurons contained the specific cholinergic marker, choline acetyltransferase, at all levels of the monkey CNS. The present observations indicate that the present CHT antibody labels cholinergic structures within the primate CNS and provides an additional marker for the investigation of cholinergic neuronal function in aging and disease.
