Interference of the noradrenergic neurotoxin DSP4 with neuronal and nonneuronal monoamine transporters

Wenge, Birger; Bönisch, Heinz

doi:10.1007/s00210-009-0459-z

Cited by 13 publications

(5 citation statements)

References 26 publications

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“…In this study, we established and characterized a novel two-hit mouse model that incorporates several common and early features of PD, including gastrointestinal inflammation and LC degeneration. Specifically, we provide a reevaluation of the DSP-4 neurotoxin model of LC injury, confirming its specificity for NE as expected based on previous studies [76][77][78][79] . We also show gastrointestinal NE depletion via DSP-4 is reversible after 4 weeks, affirming early reports of transient sympathetic denervation with DSP-4 77,79,80 .…”

Section: Discussionsupporting

confidence: 84%

Locus coeruleus injury modulates ventral midbrain neuroinflammation during DSS-induced colitis

Boles,

Holt,

Cole

et al. 2024

Preprint

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Parkinson's disease (PD) is characterized by a decades-long prodrome, consisting of a collection of non-motor symptoms that emerges prior to the motor manifestation of the disease. Of these non-motor symptoms, gastrointestinal dysfunction and deficits attributed to central norepinephrine (NE) loss, including mood changes and sleep disturbances, are frequent in the PD population and emerge early in the disease. Evidence is mounting that injury and inflammation in the gut and locus coeruleus (LC), respectively, underlie these symptoms, and the injury of these systems is central to the progression of PD. In this study, we generate a novel two-hit mouse model that captures both features, using dextran sulfate sodium (DSS) to induce gut inflammation and N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) to lesion the LC. We first confirmed the specificity of DSP-4 for central NE using neurochemical methods and fluorescence light-sheet microscopy of cleared tissue, and established that DSS-induced outcomes in the periphery, including weight loss, gross indices of gut injury and systemic inflammation, the loss of tight junction proteins in the colonic epithelium, and markers of colonic inflammation, were unaffected with DSP-4 pre-administration. We then measured alterations in neuroimmune gene expression in the ventral midbrain in response to DSS treatment alone as well as the extent to which prior LC injury modified this response. In this two-hit model we observed that DSS-induced colitis activates the expression of key cytokines and chemokines in the ventral midbrain only in the presence of LC injury and the typical DSS-associated neuroimmune is blunted by pre-LC lesioning with DSP-4. In all, this study supports the growing appreciation for the LC as neuroprotective against inflammation-induced brain injury and draws attention to the potential for NEergic interventions to exert disease-modifying effects under conditions where peripheral inflammation may compromise ventral midbrain dopaminergic neurons and increase the risk for development of PD.

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Section: Discussionsupporting

confidence: 84%

Locus coeruleus injury modulates ventral midbrain neuroinflammation during DSS-induced colitis

Boles,

Holt,

Cole

et al. 2024

Preprint

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“…DSP4 (Sigma, St Louis, MO, USA), dissolved in distilled water at 50 mM was diluted with culture media and added to cells to a final concentration of 5, 10 or 50 μM, alone or in combination with the ATM inhibitor KU55933 (10 μM, Selleckchem, USA), and/or the ATR inhibitor Nu6027 (10 μM, Santa Cruz, CA, USA) for the times as indicated in the text. The selection of the concentration of DSP4 was based on the reports about its IC50 in the literature (Boksa et al 1989, Tieu et al 1999, Wenge et al 2009) and our preliminary experiments. Only SH-SY5Y cells prior to passage 15 were used.…”

Section: Methodsmentioning

confidence: 99%

Effects of DSP4 on the Noradrenergic Phenotypes and Its Potential Molecular Mechanisms in SH-SY5Y Cells

et al. 2013

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Dopamine β-hydroxylase (DBH) and norepinephrine (NE) transporter (NET) are the noradrenergic phenotypes for their functional importance to noradrenergic neurons. It is known that in vivo N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4) treatment induces degeneration of noradrenergic terminals by interacting with NET and depleting intracellular NE. However, DSP4’s precise mechanism of action remains unclear. In this study various biochemical approaches were employed to test the hypothesis that DSP4 down-regulates the expression of DBH and NET, and to determine molecular mechanisms that may be involved. The results showed that treatment of SH-SY5Y neuroblastoma cells with DSP4 significantly decreased mRNA and protein levels of DBH and NET. DSP4-induced reduction of DBH mRNA and proteins, as well as NET proteins showed a time- and concentration-dependent manner. Flow cytometric analysis demonstrated that DSP4-treated cells were arrested predominantly in the S-phase, which was reversible. The arrest was confirmed by several DNA damage response markers (phosphorylation of H2AX and p53), suggesting that DSP4 causes replication stress which triggers cell cycle arrest via the S-phase checkpoints. Moreover, the comet assay verified that DSP4 induced single-strand DNA breaks. In summary, the present study demonstrated that DSP4 down-regulates the noradrenergic phenotypes, which may be mediated by its actions on DNA replication, leading to replication stress and cell cycle arrest. These action mechanisms of DSP4 may account for its degenerative consequence after systematic administration for animal models.

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“…monoamines and acetylcholine), drugs, toxins and other xenobiotics that are positively charged at physiologic pH (Hayer-Zillgen et al 2002;Koepsell and Endou 2004;Koepsell et al 2007;Kitamura et al 2008;Wenge and Bönisch 2009;Zolk et al 2009;Haenisch and Bönisch 2010). Since at physiological pH about 40% of all drugs are organic cations (Neuhoff et al 2003), drugs which interact with OCTs bear the potential for pharmacokinetic interactions with concomitantly administered drugs (Choi and Song 2008;Kindla et al 2009).…”

Section: Introductionmentioning

confidence: 99%

Oxybutynin and trospium are substrates of the human organic cation transporters

Wenge

Geyer

Bönisch

2011

Naunyn-Schmied Arch Pharmacol

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The muscarinic antagonists oxybutynin and trospium are used as spasmolytic agents for the treatment of overactive urinary bladder disease. Recently, it has been shown that trospium, but not oxybutynin, is a substrate of the multidrug efflux carrier P-glycoprotein, but carrier-mediated drug uptake has not been directly analysed for both drugs. However, trospium has been previously shown to exhibit inhibitory potency for the organic cation transporters (OCTs). The aim of the present study was to examine whether trospium and oxybutynin are substrates, i.e. are transported by the human OCTs (hOCT(1), hOCT(2) and hOCT(3)). Therefore, we measured total and specific (decynium-22-sensitive) uptake, and saturation kinetics of the uptake for [(3)H]oxybutynin and [(3)H]trospium in human embryonic kidney (HEK293) cells transiently transfected with the cDNA of hOCT(1), hOCT(2) or hOCT(3). In addition, we determined IC(50) values for inhibition of hOCT-mediated [(3)H]MPP(+) uptake by unlabelled trospium and oxybutynin. Total uptake of [(3)H]oxybutynin was very high in all transfected HEK293 cells and only a small portion was due to specific, decynium-22-sensitive hOCT-mediated uptake. Oxybutynin inhibited [(3)H]MPP(+) uptake by the three hOCTs with IC(50) values between 20 and 130 μM. Direct determination of transport kinetics was measurable only at hOCT(1) with K (m) of 8 μM and V (max) of 484 pmol/mg protein/min. The rank order of affinity (1/IC(50) or 1/K (m)) of specific oxybutynin uptake was hOCT(1) > hOCT(2) = hOCT(3). The observed high non-specific uptake is obviously a consequence of the high lipophilicity of this uncharged drug. Thus, hOCTs may not play a significant role for the overall pharmacokinetics and tissue distribution of oxybutynin. However, and in contrast to oxybutynin, uptake of [(3)H]trospium, an organic cation, was mainly due to carrier-mediated uptake by the three hOCTs. With IC(50) values of 18, 1.4 and 710 μM (at hOCT(1), hOCT(2) and hOCT(3), respectively) and K (m) values of 17 and 8 μM and about identical V (max) values of about 90 pmol/mg protein/min at hOCT(1) and hOCT(2), respectively; the rank order of affinity (1/IC(50) or 1/K (m)) of specific uptake of trospium was hOCT(2) > hOCT(1) > > hOCT(3). Thus, hOCTs very probably contribute to the active tubular and hepatobiliary secretion of trospium. Furthermore, hOCT(1) and hOCT(3) may be involved in the tissue uptake of this drug in the urinary bladder.

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Interference of the noradrenergic neurotoxin DSP4 with neuronal and nonneuronal monoamine transporters

Cited by 13 publications

References 26 publications

Locus coeruleus injury modulates ventral midbrain neuroinflammation during DSS-induced colitis

Locus coeruleus injury modulates ventral midbrain neuroinflammation during DSS-induced colitis

Effects of DSP4 on the Noradrenergic Phenotypes and Its Potential Molecular Mechanisms in SH-SY5Y Cells

Oxybutynin and trospium are substrates of the human organic cation transporters

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