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Hispidulin alleviated methamphetamine-induced hyperlocomotion by acting at α6 subunit-containing GABAA receptors in the cerebellum

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Abstract

Rationale

Hispidulin is a flavonoid we isolated from Clerodendrum inerme, an herb that effectively remitted a case of intractable motor tic disorders. Hispidulin was shown to be a positive allosteric modulator (PAM) of GABAA receptors, including the α6 subunit-containing subtype (α6GABAAR) that is predominantly expressed in cerebellar granule cells and insensitive to diazepam.

Objectives

We explored the action mechanism(s) of hispidulin using hyperdopaminergic mouse models induced by methamphetamine and apomorphine, based on the hyperdopaminergic nature of tic disorders.

Results

Hispidulin significantly inhibited methamphetamine-induced hyperlocomotion (MIH) at i.p. doses without affecting apomorphine-induced hyperlocomotion and stereotypy behaviors or having significant benzodiazepine-like effects (BZLE), including sedation, anxiety, and motor impairment. When given by intracerebellar (i.c.b.) microinjection, hispidulin also alleviated MIH and this effect was prevented by i.c.b. coadministration of furosemide, an α6GABAAR antagonist, and mimicked by i.c.b. Ro 15-4513, an α6GABAAR PAM. Conversely, i.c.b. diazepam did not affect MIH while it reduced MIH at i.p. doses having significant BZLE. In a screening assay for 92 neurotransmitter receptors/degradation enzymes/transporters, hispidulin displayed significant (>50 % inhibition of radiolabeled ligand binding at 10 μM) binding affinity only at the benzodiazepine binding site of GABAARs (IC50 0.73∼1.78 μM) and catecholamine-o-methyl-transferase (COMT) (IC50 1.32 μM). OR-486, a more potent COMT inhibitor than hispidulin, did not affect MIH.

Conclusions

It is suggested that hispidulin alleviates MIH via acting as a PAM of cerebellar α6GABAARs, but not through COMT inhibition or affecting dopamine receptor responsiveness. Thus, selective α6GABAAR PAMs may have the potential to be a novel treatment for hyperdopaminergic disorders.

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Abbreviations

α6GABAARs:

α6 Subunit-containing GABAA receptors

AIH:

Apomorphine-induced hyperlocomotion

AIS:

Apomorphine-induced stereotypy behaviors

CI :

Clerodendrum inerme

COMT:

Catechol-O-methyltransferase

MIH:

Methamphetamine-induced hyperlocomotion

PAM:

Positive allosteric modulator

TS:

Tourette syndrome

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Acknowledgments

This study was mainly supported by the National Research Program for Biopharmaceuticals (NSC 100-2325-B002-050, NSC 101-2325-B002-048, NSC 102-2325-B002-047, MOST 103-2325-B002-037 and MOST 104-2325-B002-010) and the research grants (MOST 103-2321-B002-035 to LCC; NSC 102-2320-B038-019-MY3 to WJH) from the National Science Council (the Ministry of Science and Technology), Taiwan, as well as by the Innovative Research Grant from National Health Research Institutes, Taiwan (NHRI-EX104-10251NI to LCC). We thank the support from Behavior Core, Neurobiology and Cognitive Center, National Taiwan University.

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Authors and Affiliations

  1. Graduate Institute of Pharmacology, College of Medicine, National Taiwan University, Taipei, Taiwan

    Yu-Hsiang Liao & Lih-Chu Chiou

  2. Department of Pharmacology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Rd., Section 1, Taipei, 100, Taiwan

    Hsin-Jung Lee & Lih-Chu Chiou

  3. Graduate Institute of Pharmacognosy, Taipei Medical University, Taipei, Taiwan

    Wei-Jan Huang

  4. Department of Pediatrics, National Taiwan University Hospital, Taipei, Taiwan

    Pi-Chuan Fan

  5. Graduate Institute of Brain and Mind Sciences, College of Medicine, National Taiwan University, Taipei, Taiwan

    Lih-Chu Chiou

  6. Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, Taiwan

    Lih-Chu Chiou

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  1. Yu-Hsiang Liao
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Correspondence to Lih-Chu Chiou.

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Supplementary Table S1

Radioligand displacement studies conducted over a broad LeadProfiling and Therapeutic area/CNS Screen by Ricerca Biosciences* (study No. AA95970) to determine the binding affinity/inhibitory activity of hispidulin at 92 receptors, channels, transporters and enzymes. (DOC 107 kb)

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Liao, YH., Lee, HJ., Huang, WJ. et al. Hispidulin alleviated methamphetamine-induced hyperlocomotion by acting at α6 subunit-containing GABAA receptors in the cerebellum. Psychopharmacology 233, 3187–3199 (2016). https://doi.org/10.1007/s00213-016-4365-z

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  • DOI: https://doi.org/10.1007/s00213-016-4365-z

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