Choline Uptake Transport Research Articles

Soluble Aβ42 Acts as Allosteric Activator of the Core Cholinergic Enzyme Choline Acetyltransferase.

Two major questions in the field of Alzheimer-type dementia remain elusive. One is the native function of amyloid-β (Aβ) peptides and the other is an early deficit in the central cholinergic network. Nevertheless, recent evidence suggests that Aβ peptides are involved in the regulation of acetylcholine (ACh) homeostasis either by allosteric activation of ACh-degrading cholinesterases or by inhibiting the high-affinity choline uptake transporter. In the current study, we report that Aβ peptides, in particular Aβ42, allosterically enhances the catalytic rate of the core-cholinergic enzyme choline acetyltransferase (ChAT), responsible for biosynthesis of ACh. Detailed in vitro enzyme kinetic analysis indicated that both soluble Aβ40 and Aβ42 enhanced the catalytic efficiency of ChAT by ∼21% and 26% at physiological concentration ranges found in human cerebrospinal fluid (CSF). Further analyses indicated that activation of ChAT by Aβ was highly specific. Intriguingly, Aβ42 exhibited an EC50 of activation potency at 10-fold lower concentrations compared to Aβ40. The activation was persistent even in the presence of a physiological Aβ 40/42 mixture ratio, expected in human CSF. In conclusion, we report for the first time that Aβ42 peptide acts as allosteric enhancers of ACh-biosynthesizing enzyme ChAT. Together with two previous observations, this points to a complex molecular cross-talk between Aβ and the enzymatic machinery involved in maintaining cellular, synaptic and extra-synaptic ACh homeostasis, warranting further investigation.

Choline acetyltransferase in the hippocampus is associated with learning strategy preference in adult male rats

One principle of the multiple memory systems hypothesis posits that the hippocampus-based and striatum-based memory systems compete for control over learning. Consistent with this notion, previous research indicates that the cholinergic system of the hippocampus plays a role in modulating the preference for a hippocampus-based place learning strategy over a striatum-based stimulus−response learning strategy. Interestingly, in the hippocampus, greater activity and higher protein levels of choline acetyltransferase (ChAT), the enzyme that synthesizes acetylcholine, are associated with better performance on hippocampus-based learning and memory tasks. With this in mind, the primary aim of the current study was to determine if higher levels of ChAT and the high-affinity choline uptake transporter (CHT) in the hippocampus were associated with a preference for a hippocampus-based place learning strategy on a task that also could be solved by relying on a striatum-based stimulus−response learning strategy. Results confirmed that levels of ChAT in the dorsal region of the hippocampus were associated with a preference for a place learning strategy on a water maze task that could also be solved by adopting a stimulus−response learning strategy. Consistent with previous studies, the current results support the hypothesis that the cholinergic system of the hippocampus plays a role in balancing competition between memory systems that modulate learning strategy preference.

Doxorubicin decreases paraquat accumulation and toxicity in Caco-2 cells

P-glycoprotein (P-gp) is an efflux pump belonging to the ATP-binding cassette transporter superfamily expressed in several organs. Considering its potential protective effects, the induction of de novo synthesis of P-gp could be used therapeutically in the treatment of intoxications by its substrates. The herbicide paraquat (PQ) is a P-gp substrate responsible for thousands of fatal intoxications worldwide that still lacks an effective antidote.The aim of the present work was to evaluate the effectiveness of such an antidote by testing whether doxorubicin (DOX), a known P-gp inducer, could efficiently protect Caco-2 cells against PQ cytotoxicity, 6h after the incubation with the herbicide, reflecting a real-life intoxication scenario. Cytotoxicity was evaluated by the MTT assay and PQ intracellular concentrations were measured by gas chromatography–ion trap-mass spectrometry (GC–IT-MS). Also, the DOX modulatory effect on choline uptake transport system was assessed by measuring the uptake of [3H]-choline.The results show that DOX exerts protective effects against PQ cytotoxicity, preventing the intracellular accumulation of the herbicide. These protective effects were not completely prevented by the incubation with the UIC2 antibody, a specific P-gp inhibitor, suggesting the involvement of alternative protection mechanisms. In fact, DOX also efficiently inhibited the choline transport system that influences PQ cellular uptake.In conclusion, in this cellular model, DOX effectively protects against PQ toxicity by inducing P-gp and through the interaction with the choline transporter, suggesting that compounds presenting this double feature of promoting the efflux and limiting the uptake of PQ could be used as effective antidotes to treat intoxications.

Effects of Cholinergic Enhancing Drugs on Cholinergic Transporters in the Brain and Peripheral Blood Lymphocytes of Spontaneously Hypertensive Rats

Cholinergic hypofunction is a trait of Alzheimer's disease and vascular dementia and countering it is one of the main therapeutic strategies available for these disorders. Cholinergic transporters control cellular mechanisms of acetylcholine (ACh) synthesis and release at presynaptic terminals. This study has assessed the influence of 4 week treatment with two different cholinergic enhancing drugs, the cholinergic precursor choline alphoscerate (alpha-glyceryl-phosphorylcholine) or the acetylcholinesterase (AChE) inhibitor galantamine on high affinity choline uptake transporter (CHT) and vesicular ACh transporter (VAChT) expression in the brain of spontaneously hypertensive rats (SHR). SHR represent an animal model of cerebrovascular injury characterized by cholinergic hypofunction. Analysis was performed by immunochemistry, ELISA and immunohistochemistry on frontal cortex, striatum and hippocampus. Immunochemical and ELISA analysis was extended to peripheral blood lymphocytes (PBL), used as a peripheral reference of changes of brain cholinergic markers. An increased expression of VAChT and CHT was observed in brain areas investigated and in PBL of SHR. The similar trend for cholinergic transporters observed in brain and PBL suggests these cells may represent a marker of brain cholinergic transporters. Treatment with choline alphoscerate increased CHT and to a greater extent VAChT expression. Treatment with galantamine countered the increase of CHT and VAChT. The different activity of the cholinergic precursor and of the AChE inhibitor on parameters investigated is likely related to their mechanism of action. Choline alphoscerate increases ACh synthesis and release. This requires an augmentation of systems regulating neurotransmitter uptake and storage. The effect of choline alphoscerate on CHT and VAChT observed in this study suggests an improved synaptic efficiency elicited by the compound. The AChE inhibitor slows-down ACh degradation in the synaptic cleft. A greater availability of neurotransmitter elicited by galantamine counters the enhanced activity of cholinergic transporters compensating cholinergic deficits. Differences in the activity of the cholinergic precursor and AChE inhibitor investigated on CHT and VAChT suggests that association between choline alphoscerate and AChE/cholinesterase inhibitors may represent a strategy for potentiating deficient cholinergic neurotransmission worthwhile of being investigated in clinical trials.

Synthesis and characterization of luminescent cadmium selenide/zinc selenide/zinc sulfide cholinomimetic quantum dots

Luminescent quantum dots conjugated with highly selective molecular recognition ligands are widely used for targeting and imaging biological structures. In this paper, water soluble cholinomimetic cadmium selenide (core), zinc selenide/zinc sulfide (shell) quantum dots were synthesized for targeting cholinergic sites. Cholinomimetic specificity was incorporated by conjugation of the quantum dots to an aminated analogue of hemicholinium-15, a well known competitive inhibitor of the high affinity choline uptake transporter. Detailed evaluation of the nanocrystal synthesis and characterization of the final product was conducted by (1)H and (31)P NMR, absorption and emission spectroscopy, as well as transmission electron microscopy.

Choline transporters, cholinergic transmission and cognition

Cholinergic projections to the cortex and hippocampus mediate fundamental cognitive processes. The capacity of the high-affinity choline uptake transporter (CHT) to import choline from the extracellular space to presynaptic terminals is essential for normal acetylcholine synthesis and therefore cholinergic transmission. The CHT is highly regulated, and the cellular mechanisms that modulate its capacity show considerable plasticity. Recent evidence links changes in CHT capacity with the ability to perform tasks that tax attentional processes and capacities. Abnormal regulation of CHT capacity might contribute to the cognitive impairments that are associated with neurodegenerative and neuropsychiatric disorders. Therefore, the CHT might represent a productive target for the development of new pharmacological treatments for these conditions.

The presynaptic component of a cholinergic mechanism in the carotid body chemotransduction of hypoxia in the cat.

In the past we have shown that cholinergic blockers given together or singly could greatly reduce the carotid body’s response to a perfusion of hypoxic Krebs Ringer bicarbonate solution (KRB) (Fitzgerald & Shirahata, 1994; Fitzgerald et al, 1995). The operating model in those studies was transmission in the superior cervical ganglion. That is, the Type I cell of the carotid body was analogous to the presynaptic neuron; it contained acetylcholine (ACh). In fact the Type I cell has been identified as the source of ACh in several different studies. The synthesizing enzyme, choline acetyltransferase (ChAT), has been located there (Ballard & Jones, 1972; Wang et al, 1989). Fidone et al (1977) have located the high affinity sodium-dependent choline uptake transporter in the Type I cells. They have also reported that carotid bodies which had been denervated contained as much ACh as their intact counterparts (Fidone et al, 1976), suggesting that the extensive innervation in the carotid body and the very few ganglia reported to be there contribute an immeasurably small amount of ACh. And we (Goldberg et al, 1978) analyzed the carotid sinus nerve directly to find no appreciable amount of ACh in it.