Severe neonatal hyperbilirubinemia has been known to cause the clinical syndrome of kernicterus and a milder one the syndrome of bilirubin-induced neurologic dysfunction (BIND). BIND clinically manifests itself after the neonatal period as developmental delay, cognitive impairment, and related behavioral and psychiatric disorders. The complete picture of BIND is not clear.
The Gunn rat is a mutant strain of the Wistar rat with the BIND phenotype, and it demonstrates abnormal behavior. We investigated serotonergic dysfunction in Gunn rats by pharmacological analyses and ex vivo neurochemical analyses.
Ketanserin, the 5-HT2AR antagonist, normalizes hyperlocomotion of Gunn rats. Both serotonin and its metabolites in the frontal cortex of Gunn rats were higher in concentrations than in control Wistar rats. The 5-HT2AR mRNA expression was downregulated without alteration of the protein abundance in the Gunn rat frontal cortex. The TPH2 protein level in the Gunn rat raphe region was significantly higher than that in the Wistar rat.
It would be of value to be able to postulate that a therapeutic strategy for BIND disorders would be the restoration of brain regions affected by the serotonergic dysfunction to normal operation to prevent before or to normalize after onset of BIND manifestations.
We demonstrated serotonergic dysregulation underlying hyperlocomotion in Gunn rats. This finding suggests that a therapeutic strategy for bilirubin-induced neurologic dysfunction (BIND) would be the restoration of brain regions affected by the serotonergic dysfunction to normal operation to prevent before or to normalize after the onset of the BIND manifestations.
Ketanserin normalizes hyperlocomotion of Gunn rats.
To our knowledge, this is the first study to demonstrate a hyperlocomotion link to serotonergic dysregulation in Gunn rats.
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Richard, J., Avroy, A. & Michele, C. Fanaroff and Martin’s Neonatal-Perinatal Medicine 2-Volume Set 11th edn (Elsevier, 2019).
Wu, X. J., Zhong, D. N., Xie, X. Z., Ye, D. Z. & Gao, Z. Y. UGT1A1 gene mutations and neonatal hyperbilirubinemia in Guangxi Heiyi Zhuang and Han populations. Pediatr. Res. 78, 585–588 (2015).
Nguyen, T. T., Zhao, W., Yang, X. & Zhong, D. N. The relationship between hyperbilirubinemia and the promoter region and first exon of UGT1A1 gene polymorphisms in Vietnamese newborns. Pediatr. Res. 88, 940–944 (2020).
Wang, J., Yin, J., Xue, M., Lyu, J. & Wan, Y. Roles of UGT1A1 Gly71Arg and TATA promoter polymorphisms in neonatal hyperbilirubinemia: a meta-analysis. Gene 736, 144409 (2020).
Long, J., Zhang, S., Fang, X., Luo, Y. & Liu, J. Neonatal hyperbilirubinemia and Gly71Arg mutation of UGT1A1 gene: a Chinese case-control study followed by systematic review of existing evidence. Acta Paediatr. 100, 966–971 (2011).
Dalman, C. & Cullberg, J. Neonatal hyperbilirubinaemia-a vulnerability factor for mental disorder? Acta Psychiatr. Scand. 100, 469–471 (1999).
Maimburg, R. D. et al. Neonatal jaundice: a risk factor for infantile autism? Paediatr. Perinat. Epidemiol. 22, 562–568 (2008).
Wei, C. C. et al. Neonatal jaundice and increased risk of attention-deficit hyperactivity disorder: a population-based cohort study. J. Child Psychol. Psychiatry 56, 460–467 (2015).
Miyaoka, T., Seno, H., Maeda, T., Itoga, M. & Horiguchi, J. Schizophrenia-associated idiopathic unconjugated hyperbilirubinemia (Gilbert’s syndrome): 3 case reports. J. Clin. Psychiatry 61, 299–300 (2000).
Miyaoka, T. et al. Schizophrenia-associated idiopathic unconjugated hyperbilirubinemia (Gilbert’s syndrome). J. Clin. Psychiatry 61, 868–871 (2000).
Amin, S. B., Smith, T. & Timler, G. Developmental influence of unconjugated hyperbilirubinemia and neurobehavioral disorders. Pediatr. Res. 85, 191–197 (2019).
Schutta, H. S. & Johnson, L. Bilirubin encephalopathy in the Gunn rat: a fine structure study of the cerebellar cortex. J. Neuropathol. Exp. Neurol. 26, 377–396 (1967).
Gunn, C. K. Hereditary acholuric jaundice in the rat. Can. Med. Assoc. J. 50, 230–237 (1944).
Daood, M. J., Hoyson, M. & Watchko, J. F. Lipid peroxidation is not the primary mechanism of bilirubin-induced neurologic dysfunction in jaundiced Gunn rat pups. Pediatr. Res. 72, 455–459 (2012).
Schutta, H. S. & Johnson, L. Clinical signs and morphologic abnormalities in Gunn rats treated with sulfadimethoxine. J. Pediatr. 75, 1070–1079 (1969).
Hayashida, M. et al. Hyperbilirubinemia-related behavioral and neuropathological changes in rats: a possible schizophrenia animal model. Prog. Neuropsychopharmacol. Biol. Psychiatry 33, 581–588 (2009).
Liaury, K. et al. Minocycline improves recognition memory and attenuates microglial activation in Gunn rat: a possible hyperbilirubinemia-induced animal model of schizophrenia. Prog. Neuropsychopharmacol. Biol. Psychiatry 50, 184–190 (2014).
Tsuchie, K. et al. The effects of antipsychotics on behavioral abnormalities of the Gunn rat (unconjugated hyperbilirubinemia rat), a rat model of schizophrenia. Asian J. Psychiatr. 6, 119–123 (2013).
Stanford, J. A. et al. Hyperactivity in the Gunn rat model of neonatal jaundice: age-related attenuation and emergence of gait deficits. Pediatr. Res. 77, 434–439 (2015).
Leysen, J. E. et al. Biochemical profile of risperidone, a new antipsychotic. J. Pharmacol. Exp. Ther. 247, 661–670 (1988).
Laduron, P. M., Janssen, P. F. & Leysen, J. E. In vivo binding of [3H]ketanserin on serotonin S2-receptors in rat brain. Eur. J. Pharmacol. 81, 43–48 (1982).
Oh-Nishi, A., Saji, M., Furudate, S. I. & Suzuki, N. Dopamine D(2)-like receptor function is converted from excitatory to inhibitory by thyroxine in the developmental hippocampus. J. Neuroendocrinol. 17, 836–845 (2005).
Oh-Nishi, A., Obayashi, S., Sugihara, I., Minamimoto, T. & Suhara, T. Maternal immune activation by polyriboinosinic-polyribocytidilic acid injection produces synaptic dysfunction but not neuronal loss in the hippocampus of juvenile rat offspring. Brain Res. 1363, 170–179 (2010).
Iyanagi, T., Watanabe, T. & Uchiyama, Y. The 3-methylcholanthrene-inducible UDP-glucuronosyltransferase deficiency in the hyperbilirubinemic rat (Gunn rat) is caused by a -1 frameshift mutation. J. Biol. Chem. 264, 21302–21307 (1989).
Malkova, N. V., Gallagher, J. J., Yu, C. Z., Jacobs, R. E. & Patterson, P. H. Manganese-enhanced magnetic resonance imaging reveals increased DOI-induced brain activity in a mouse model of schizophrenia. Proc. Natl Acad. Sci. USA 111, E2492–E2500 (2014).
Ogata, M., Akita, H. & Ishibashi, H. Behavioral responses to anxiogenic tasks in young adult rats with neonatal dopamine depletion. Physiol. Behav. 204, 10–19 (2019).
Kalueff, A. V. & Tuohimaa, P. Grooming analysis algorithm for neurobehavioural stress research. Brain Res. Brain Res. Protoc. 13, 151–158 (2004).
Paxinos, G. & Watson, C. The Rat Brain IN Stereotaxic Coordinates 6th edn (Elsevier, 2007).
Shimoyama, M. et al. The Rat Genome Database 2015: genomic, phenotypic and environmental variations and disease. Nucleic Acids Res. 43, D743–D750 (2015).
Hayashida, M. et al. Parvalbumin-positive GABAergic interneurons deficit in the hippocampus in Gunn rats: a possible hyperbilirubinemia-induced animal model of schizophrenia. Heliyon 5, e02037 (2019).
Pazos, A., Cortes, R. & Palacios, J. M. Quantitative autoradiographic mapping of serotonin receptors in the rat brain. II. Serotonin-2 receptors. Brain Res. 346, 231–249 (1985).
NIMH. NIMH Psychoactive Drug Screening Program. https://pdsp.unc.edu/ (2017).
Hensler, J. G. Serotonergic modulation of the limbic system. Neurosci. Biobehav. Rev. 30, 203–214 (2006).
Bjorklund, A. & Dunnett, S. B. Dopamine neuron systems in the brain: an update. Trends Neurosci. 30, 194–202 (2007).
Shapiro, S. M. Definition of the clinical spectrum of kernicterus and bilirubin-induced neurologic dysfunction (BIND). J. Perinatol. 25, 54–59 (2005).
Puig, M. V. & Gulledge, A. T. Serotonin and prefrontal cortex function: neurons, networks, and circuits. Mol. Neurobiol. 44, 449–464 (2011).
Aghajanian, G. K. & Marek, G. J. Serotonin induces excitatory postsynaptic potentials in apical dendrites of neocortical pyramidal cells. Neuropharmacology 36, 589–599 (1997).
Aghajanian, G. K. & Marek, G. J. Serotonin, via 5-HT2A receptors, increases EPSCs in layer V pyramidal cells of prefrontal cortex by an asynchronous mode of glutamate release. Brain Res. 825, 161–171 (1999).
Kanno, H. et al. Effect of yokukansan, a traditional Japanese medicine, on social and aggressive behaviour of para-chloroamphetamine-injected rats. J. Pharm. Pharmacol. 61, 1249–1256 (2009).
Ninan, I. & Kulkarni, S. K. 5-HT2A receptor antagonists block MK-801-induced stereotypy and hyperlocomotion. Eur. J. Pharmacol. 358, 111–116 (1998).
Grillner, S., & El Manira, A. Current principles of motor control, with special reference to vertebrate locomotion. Physiol. Rev. 100, 271–320 (2020).
Flaive, A., Fougere, M., van der Zouwen, C. I. & Ryczko, D. Serotonergic modulation of locomotor activity from basal vertebrates to mammals. Front. Neural Circuits 14, 590299 (2020).
Ryczko, D. & Dubuc, R. Dopamine and the brainstem locomotor networks: from lamprey to human. Front. Neurosci. 11, 295 (2017).
Butcher, R. E., Stutz, R. M. & Berry, H. K. Behavioral abnormalities in rats with neonatal jaundice. Am. J. Ment. Defic. 75, 755–759 (1971).
Swenson, R. M. & Jew, J. Y. Learning deficits and brain monoamines in rats with congenital hyperbilirubinemia. Exp. Neurol. 76, 447–456 (1982).
Jew, J. Y. & Sandquist, D. CNS changes in hyperbilirubinemia. Functional implications. Arch. Neurol. 36, 149–154 (1979).
Furuya, M. et al. Yokukansan promotes hippocampal neurogenesis associated with the suppression of activated microglia in Gunn rat. J. Neuroinflammation 10, 145 (2013).
Limoa, E. et al. Electroconvulsive shock attenuated microgliosis and astrogliosis in the hippocampus and ameliorated schizophrenia-like behavior of Gunn rat. J. Neuroinflammation 13, 230 (2016).
Arauchi, R. et al. Gunn rats with glial activation in the hippocampus show prolonged immobility time in the forced swimming test and tail suspension test. Brain Behav. 8, e01028 (2018).
Scotton, W. J., Hill, L. J., Williams, A. C. & Barnes, N. M. Serotonin syndrome: pathophysiology, clinical features, management, and potential future directions. Int. J. Tryptophan Res. 12, 1178646919873925 (2019).
Lozada, L. E. et al. Association of autism spectrum disorders with neonatal hyperbilirubinemia. Glob. Pediatr. Health 2, 2333794X15596518 (2015).
Biederman, J. & Faraone, S. V. Attention-deficit hyperactivity disorder. Lancet 366, 237–248 (2005).
Ribases, M. et al. Exploration of 19 serotoninergic candidate genes in adults and children with attention-deficit/hyperactivity disorder identifies association for 5HT2A, DDC and MAOB. Mol. Psychiatry 14, 71–85 (2009).
Tanaka, M. et al. Brain hyperserotonemia causes autism-relevant social deficits in mice. Mol. Autism 9, 60 (2018).
Shen, H. W. et al. Regional differences in extracellular dopamine and serotonin assessed by in vivo microdialysis in mice lacking dopamine and/or serotonin transporters. Neuropsychopharmacology 29, 1790–1799 (2004).
Bonnier, C., Nassogne, M. C. & Evrard, P. Ketanserin treatment of Tourette’s syndrome in children. Am. J. Psychiatry 156, 1122–1123 (1999).
Muller, N., Schiller, P. & Ackenheil, M. Coincidence of schizophrenia and hyperbilirubinemia. Pharmacopsychiatry 24, 225–228 (1991).
We are grateful to Professor emeritus Tadakazu Maeda (Kitasato Univ.) and Dr. Nobuyuki Suzuki (neuroscientist) for giving us advice and to Riho Murai (Shimane Univ.), Kohei Ueda (Shimane Univ.), and Ayumi Fujiwara (Shimane Univ.) for research assistances. Funding came from KAKENHI 19K17363 (to S.M.), from the Ministry of Education, Culture, Sports, Science and Technology (MEXT).
Shimane University has submitted a patent application (PCT/JP2020/017552) for the use of the compound described in this paper. M.I. has received grants from Research for Promotion of Cancer Control Programs during the conduct of the study. He has received lecture fees from Technomics, Fuji Keizai, Novartis, Yoshitomiyakuhin, Pfizer, MSD, Meiji Seika Pharma, Eisai, Otsuka, Sumitomo Dainippon Pharma, Mochida, Janssen, Takeda, and Eli Lilly. The institution of M.I. received grants or research support from Otsuka, Eisai, Daiichi-Sankyo, Pfizer, Astellas, MSD, Takeda, Fujifilm, Shionogi, and Mochida. A.O.-N. is listed as an inventor of this patent. A.O.-N. is CEO & CTO of RESVO Inc. and has >5% of RESVO Inc. shares but had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. This does not alter our adherence to Pediatric Research publication policy. Other authors have no competing interests to declare.
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Miura, S., Tsuchie, K., Fukushima, M. et al. Normalizing hyperactivity of the Gunn rat with bilirubin-induced neurological disorders via ketanserin. Pediatr Res 91, 556–564 (2022). https://doi.org/10.1038/s41390-021-01446-1