Abstract
Auditory dysfunction, including hypersensitivity and tinnitus, is a common symptom of autism spectrum disorder (ASD). Prenatal exposure to the antiseizure medication valproic acid (VPA) significantly increases the risk of ASD in humans and similar exposure is utilized as an animal model of ASD in rodents. Animals exposed to VPA in utero have abnormal activity in their auditory cortex in response to sounds, fewer neurons, abnormal neuronal morphology, reduced expression of calcium-binding proteins, and reduced ascending projections to the central nucleus of the inferior colliculus. Unfortunately, these previous studies of central auditory circuits neglect the medial geniculate (MG), which serves as an important auditory relay from the midbrain to the auditory cortex. Here, we examine the structure and connectivity of the medial geniculate (MG) in rats prenatally exposed to VPA. Our results indicate that VPA exposure results in significantly smaller and fewer neurons in the ventral and medial nuclei of the MG. Furthermore, injections of the retrograde tract tracer fluorogold (FG) in the MG result in significantly fewer FG+ neurons in the inferior colliculus, superior olivary complex, and ventral cochlear nucleus. Together, we interpret these findings to indicate that VPA exposure results in hypoplasia throughout the auditory circuits and that VPA has a differential impact on some long-range axonal projections from brainstem centers to the thalamus. Together, our findings support the widespread impact of VPA on neurons and sensory circuits in the developing brain.
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Abbreviations
- ASD:
-
Autism spectrum disorder
- AVCN:
-
Anterior ventral cochlear nucleus
- Cb:
-
Cerebellum
- CL:
-
Contralateral
- CN:
-
Cochlear nucleus
- CNIC:
-
Central nucleus of the inferior colliculus
- D:
-
Dorsal
- DNLL:
-
Dorsal nucleus of the lateral lemniscus
- DMW :
-
Dorsal medial wedge
- E:
-
Embryonic
- FG:
-
Fluorogold
- FG+ :
-
Fluorogold positive
- IC:
-
Inferior colliculus
- IL:
-
Ipsilateral
- L:
-
Lateral
- ll:
-
Lateral lemniscus
- LSO:
-
Lateral superior olive
- M:
-
Medial
- MG:
-
Medial geniculate
- mMG:
-
Medial nucleus of the medial geniculate
- MNTB:
-
Medial nucleus of the trapezoid body
- MSO:
-
Medial superior olive
- NLL:
-
Nuclei of the lateral lemniscus
- NTR:
-
Neurotrace red
- P:
-
Postnatal
- PB:
-
Phosphate buffer
- SD:
-
Standard deviation
- SOC:
-
Superior olivary complex
- SPON:
-
Superior paraolivary nucleus
- STN:
-
Spinal trigeminal nucleus
- stt:
-
Spinal trigeminal tract
- tz:
-
Trapezoid body
- V:
-
Vestibular nerve
- VCN:
-
Ventral cochlear nucleus
- vMG:
-
Ventral nucleus of the medial geniculate
- VPA:
-
Valproic acid
References
Allen DA (1988) Autistic spectrum disorders: clinical presentation in preschool children. J Child Neurol 3:48–56
American Psychiatric Association (2013) Diagnostic and statistical manual of mental disorders. American Psychiatric Association, Washington
Anderson LA, Malmierca MS, Wallace MN, Palmer AR (2006) Evidence for a direct, short latency projection from the dorsal cochlear nucleus to the auditory thalamus in the guinea pig. Eur J Neurosci 24(2):491–498
Anomal RF, de Villers-Sidani E, Brandão JA, Diniz R, Costa MR, Romcy-Pereira RN (2015) Impaired processing in the primary auditory cortex of an animal model of autism. Front Syst Neurosci 1(9):158
Aparicio MA, Saldaña E (2014) The dorsal tectal longitudinal column (TLCd): a second longitudinal column in the paramedian region of the midbrain tectum. Brain Struct Funct 219(2):607–630
Ardinger HH, Atkin JF, Blackston RD, Elsas LJ, Clarren SK, Livingstone S, Flannery DB, Pellock JM, Harrod MJ, Lammer EJ (1988) Verification of the fetal valproate syndrome phenotype. Am J Med Genet 29(1):171–185
Bajo VM, Merchán MA, López DE, Roullier EM (1993) Neuronal morphology and efferent projections of the dorsal nucleus of the lateral lemniscus of the cat. J Compar Neurol 334:241–262
Berkley KJ (1980) Spatial relationships between the terminations of somatic sensory and motor pathways in the rostral brainstem of cats and monkeys I. Ascending somatic sensory inputs to lateral diencephalon. J Comp Neurol 193:283–317
Berrebi A, Auxiliadora AM, Jin Y, Sloan DM, Gómez-Alvarez M, Bernardo FJ, Saldaña E (2012) Direct Projections from Multiple Nuclei of the Superior Olivary Complex to the Medial Geniculate Body of the Thalamus in the Rat. Abstract #250. Association for Research in Otolaryngology, 2012 Mid-Winter Meeting.
Blum PS, Day MI, Carpenter MB, Gilman S (1979) Thalamic components of the ascending vestibular system. Exp Neurol 54:587–603
Bolton PF, Golding J, Emond A, Steer CD (2012) Autism spectrum disorder and autistic traits in the Avon Longitudinal Study of Parents and Children: precursors and early signs. J Am Acad Child Adolesc Psychiatry 51(3):249-260.e25
Bromley RL, Mawer GE, Briggs M, Cheyne C, Clayton-Smith J, García-Fiñana M, Kneen R, Lucas SB, Shallcross R, Baker GA, Liverpool and Manchester Neurodevelopment Group (2013) The prevalence of neurodevelopmental disorders in children prenatally exposed to antiepileptic drugs. J Neurol Neurosurg Psychiatry 84(6):637–643
CDC.gov (2019). https://www.cdc.gov/ncbddd/autism/index.html. Accessed 30 Jan 2019
Chelini G, Zerbi V, Cimino L et al (2019) Aberrant Somatosensory Processing and connectivity in mice lacking Engrailed-2. J Neurosci 39(8):1525–1538
Christensen J, Gronborg TK, Sorensen MJ (2013) Prenatal valproate exposure and risk of autism spectrum disorders and childhood autism. JAMA 309(16):1696–1703
Danesh AA, Lang D, Kaf W, Andreassen WD, Scott J, Eshraghi AA (2015) Tinnitus and hyperacusis in autism spectrum disorders with emphasis on high functioning individuals diagnosed with Asperger’s Syndrome. Int J Pediatr Otorhinolaryngol 79(10):1683–1688
DiLiberti JH, Farndon PA, Dennis NR, Curry CJ (1984) The fetal valproate syndrome. Am J Med Genet 19(3):473–481
Dubiel A, Kulesza RJ Jr (2016) Prenatal valproic acid exposure disrupts tonotopic c-Fos expression in the rat brainstem. Neuroscience 324:511–523
Engineer CT, Centanni TM, Im KW, Borland MS, Moreno NA, Carraway RS, Wilson LG, Kilgard MP (2014) Degraded auditory processing in a rat model of autism limits the speech representation in non-primary auditory cortex. Dev Neurobiol 74(10):972–986
Foran L, Blackburn K, Kulesza RJ (2017) Auditory hindbrain atrophy and anomalous calcium binding protein expression after neonatal exposure to monosodium glutamate. Neuroscience 344:406–417
Gandal MJ, Edgar JC, Ehrlichman RS, Mehta M, Roberts TP, Siegel SJ (2010) Validating γ oscillations and delayed auditory responses as translational biomarkers of autism. Biol Psychiatry 68(12):1100–1106
Gomes E, Pedroso FS, Wagner MB (2008) Auditory hypersensitivity in the autistic spectrum disorder. Pro Fono 20(4):279–284
González-Hernández TH, Galindo-Mireles D, Castaneyra-Perdomo A, Ferres-Torres R (1991) Divergent projections of projecting neurons of the inferior colliculus to the medial geniculate body and the contralateral inferior colliculus in the rat. Hear Res 52:17–21
Graham J (1977) An autoradiographic study of the efferent connections of the superior colliculus in the cat. J Comp Neurol 173:629–654
Greenspan SI, Wieder S (1997) Developmental patterns and outcomes in infants and children with disorders in relating and communicating: a chart review of 200 cases of children with autistic spectrum diagnoses. J Dev Learn Disord 1:87–141
Güveli BT, Rosti RÖ, Güzeltaş A, Tuna EB, Ataklı D, Sencer S, Yekeler E, Kayserili H, Dirican A, Bebek N, Baykan B, Gökyiğit A, Gürses C (2017) Teratogenicity of antiepileptic drugs. Clin Psychopharmacol Neurosci 15(1):19–27
Hou Q, Wang Y, Li Y, Chen D, Yang F, Wang S (2018) A developmental study of abnormal behaviors and altered gabaergic signaling in the vpa-treated rat model of autism. Front Behav Neurosci 12:182
Ito T, Oliver DL (2010) Origins of glutamatergic terminals in the inferior colliculus identified by retrograde transport and expression of VGLUT1 and VGLUT2 genes. Front Neuroanat 4:135
Kelly JB, Buckthought AD, Kidd SA (1998) Monaural and binaural response properties of single neurons in the rat’s dorsal nucleus of the lateral lemniscus. Hear Res 122(1–2):25–40
Kelly JB, van Adel BA, Ito M (2009) Anatomical projections of the nuclei of the lateral lemniscus in the albino rat (Rattus norvegicus). J Comp Neurol 512(4):573–593
Kolodny T, Schallmo MP, Gerdts J, Edden RAE, Bernier RA, Murray SO (2020) Concentrations of cortical GABA and glutamate in young adults with autism spectrum disorder. Autism Res. https://doi.org/10.1002/aur.2300
Kondo HM, Lin IF (2020) Excitation-inhibition balance and auditory multistable perception are correlated with autistic traits and schizotypy in a non-clinical population. Sci Rep 10(1):8171
Konigsmark BW (1970) Methods for counting of neurons. In: Nauta WJH, Ebbesson SOE (eds) Contemporary research methods in neuroanatomy. Springer, Heidelberg, pp 315–338
Koren G, Nava-Ocampo AA, Moretti ME, Sussman R, Nulman I (2006) Major malformations with valproic acid. Can Fam Physician 52:441–447
Kulesza RJ Jr (2007) Cytoarchitecture of the human superior olivary complex: medial and lateral superior olive. Hear Res 225(1–2):80–90
Kulesza RJ, Berrebi AS (2000) Superior paraolivary nucleus of the rat is a gabaergic nucleus. J Assoc Res Otolaryngol 4:255–269
Kulesza RJ, Mangunay K (2008) Morphological features of the medial superior olive in autism. Brain Res 1200:132–137
Kulesza RJ, Viñuela A, Saldaña E, Berrebi AS (2002) Unbiased stereological estimates of neuron number in subcortical auditory nuclei of the rat. Hear Res 168(1–2):12–24
Kulesza RJ, Lukose R, Stevens LV (2011) Malformation of the human superior olive in autistic spectrum disorders. Brain Res 1367:360–371
Kumamaru E, Egashira Y, Takenaka R, Takamori S (2014) Valproic acid selectively suppresses the formation of inhibitory synapses in cultured cortical neurons. Neurosci Lett 569:142–147
LeDoux JE, Ruggiero DA, Forest R, Stornetta R, Reis DJ (1987) Topographic organization of convergent projections to the thalamus from the inferior colliculus and spinal cord in the rat. J Compar Neurol 264:123–146
Lukose R, Schmidt E, Wolski TP Jr, Murawski NJ, Kulesza RJ (2011) Malformation of the superior olivary complex in an animal model of autism. Brain Res 1398:102–112
Lukose R, Brown K, Barber CM, Kulesza RJ (2013) Quantification of the stapedial reflex reveals delayed responses in autism. Autism Res 6(5):344–353
Lukose R, Beebe K, Kulesza RJ (2015) Organization of the human superior olivary complex in 15q duplication syndromes and autism spectrum disorders. Neuroscience 286:216–230
Mabunga DF, Gonzales EL, Kim JW, Kim KC, Shin CY (2015) Exploring the validity of valproic acid animal model of autism. Exp Neurobiol 24(4):285–300
Main S, Kulesza RJ (2017) Repeated prenatal exposure to valproic acid results in cerebellar hypoplasia and ataxia. Neuroscience 340:34–47
Malmierca M (2015) Auditory system. In: Paxinos G (ed) The rat nervous system. Academic Press, New York
Malmierca MS, Merchan MA, Henkel CK, Oliver DL (2002) Direct projections from cochlear nuclear complex to auditory thalamus in the rat. J Neurosci 22:10891–10897
Mansour Y, Mangold S, Chosky D, Kulesza RJ (2019a) Auditory midbrain hypoplasia and dysmorphology after prenatal valproic acid exposure. Neuroscience 396:79–93
Mansour Y, Altaher W, Kulesza RJ Jr (2019b) Characterization of the human central nucleus of the inferior colliculus. Hear Res 377:234–246
Mansour Y, Kulesza R (2020) Three dimensional reconstructions of the superior olivary complex from children with autism spectrum disorder. Hear Res. 393:107974
Mansour Y, Ahmed S, Kulesza RJ (2019c) Hypoplasia and reduced ascending axonal projections to the auditory thalamus after in utero exposure to valproic acid. Tom Ridge Environmental Center Research Symposium, Erie
Márquez-Legorreta E, Horta-Júnior AJ, Berrebi AS, Saldaña E (2016) Organization of the zone of transition between the pretectum and the thalamus, with emphasis on the pretectothalamic lamina. Front Neuroanat 10:82
Mellott JG, Foster NL, Ohl AP, Schofield BR (2014) Excitatory and inhibitory projections in parallel pathways from the inferior colliculus to the auditorythalamus. Front Neuroanat. 8:124
Moon BS, Lu W, Park HJ (2018) Valproic acid promotes the neuronal differentiation of spiral ganglion neural stem cells with robust axonal growth. Biochem Biophys Res Commun 503(4):2728–2735
Moore SJ, Turnpenny P, Quinn A, Glover S, Lloyd DJ, Montgomery T, Dean JC (2000) A clinical study of 57 children with fetal anticonvulsant syndromes. J Med Genet 37(7):489–497
Nair A, Treiber JM, Shukla DK, Shih P, Müller RA (2013) Impaired thalamocortical connectivity in autism spectrum disorder: a study of functional and anatomical connectivity. Brain 36(6):1942–1955
O’Connor K (2012) Auditory processing in autism spectrum disorder: a review. Neurosci Biobehav Rev 36(2):836–854
Olexová L, Štefánik P, Kršková L (2016) Increased anxiety-like behaviour and altered GABAergic system in the amygdala and cerebellum of VPA rats. An animal model of autism. Neurosci Lett 629:9–14
Oliver DL (1984) Neuron types in the central nucleus of the inferior coIIicuius that project to the mediai geniculate body. Neuroscience 11:409–424
Paxinos G, Watson C (2007) The rat brain in stereotaxic coordinates. Academic Press, London
Peruzzi D, Bartlett E, Smith PH, Oliver DL (1997) A monosynaptic GABAergic input from the inferior colliculus to the medial geniculate body in rat. J Neurosci 17:3766–3777
Rasalam AD, Hailey H, Williams JH, Moore SJ, Turnpenny PD, Lloyd DJ, Dean JC (2005) Characteristics of fetal anticonvulsant syndrome associated autistic disorder. Dev Med Child Neurol 47(8):551–555
Rodier PM, Ingram JL, Tisdale B, Nelson S, Romano J (1996) Embryological origin for autism: developmental anomalies of the cranial nerve motor nuclei. J Comp Neurol 370(2):247–261
Roucoux-Hanus M, Boisacq-Schepens N (1977) Ascending vestibular projections: further results at cortical and thalamic levels in the cat. Expl Brain Res 29:283–292
Rueden CT, Schindelin J, Hiner MC, DeZonia BE, Walter AE, Arena ET, Eliceiri KW (2017) Image J2: ImageJ for the next generation of scientific image data. BMC Bioinform 18(1):529
Ruby K, Falvey K, Kulesza RJ (2015) Abnormal neuronal morphology and neurochemistry in the auditory brainstem of Fmr1 knockout rats. Neuroscience. 303:285–98
Saldaña E, Aparicio MA, Fuentes-Santamaría V, Berrebi AS (2009) Connections of the superior paraolivary nucleus of the rat: projections to the inferior colliculus. Neuroscience 163(1):372–387
Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9(7):676–682
Schneider T, Roman A, Basta-Kaim A, Kubera M, Budziszewska B, Schneider K, Przewłocki R (2008) Gender-specific behavioral and immunological alterations in an animal model of autism induced by prenatal exposure to valproic acid. Psychoneuroendocrinology 33(6):728–740
Schofield BR, Mellott JG, Motts SD (2014a) Subcollicular projections to the auditory thalamus and collateral projections to the inferior colliculus. Front Neuroanat 8:70
Schofield BR, Motts SD, Mellott JG, Foster NL (2014b) Projections from the dorsal and ventral cochlear nuclei to the medial geniculate body. Front Neuroanat 8:10
Smith A, Storti S, Lukose R, Kulesza RJ Jr (2019) Structural and functional aberrations of the auditory brainstem in autism spectrum disorder. J Am Osteopath Assoc 119(1):41–50
Strominger NL (1973) The origins, course and distribution of the dorsal and intermediate acoustic striae in the rhesus monkey. J Comp Neurol 147:209–233
Strominger NL, Strominger AI (1971) Ascending brain stem projections of the anteroventral cochlear nucleus in the rhesus monkey. J Comp Neurol 143:217–242
Tamura R, Kitamura H, Endo T, Hasegawa N, Someya T (2010) Reduced thalamic volume observed across different subgroups of autism spectrum disorders. Psychiatry Res 184(3):186–188
Tang S, Powell EM, Zhu W, Lo FS, Erzurumlu RS, Xu S (2019) Altered forebrain functional connectivity and neurotransmission in a kinase-inactive met mouse model of autism. Mol Imaging 18:1536012118821034
Thompson CK, Brenowitz EA (2005) Seasonal change in neuron size and spacing but not neuronal recruitment in a basal ganglia nucleus in the avian song control system. J Comp Neurol 481(3):276–283
Tomasi D, Volkow ND (2019) Reduced local and increased long-range functional connectivity of the thalamus in autism spectrum disorder. Cereb Cortex 29(2):573–585. https://doi.org/10.1093/cercor/bhx340
Tomchek SD, Dunn W (2007) Sensory processing in children with and without autism: a comparative study using the short sensory profile. Am J Occup Ther 61(2):190–200
Tsatsanis KD, Rourke BP, Klin A, Volkmar FR, Cicchetti D, Schultz RT (2003) Reduced thalamic volume in high-functioning individuals with autism. Biol Psychiatry 53(2):121–129
Vinuela A, Aparicio MA, Berrebi AS, Saldaña E (2011) Connections of the Superior paraolivary nucleus of the rat: II Reciprocal connections with the tectal longitudinal column. Front Neuroanat 5:1
Wei R, Li Q, Lam S, Leung J, Cheung C, Zhang X, Sham PC, Chua SE, McAlonan GM (2016) A single low dose of valproic acid in late prenatal life alters postnatal behavior and glutamic acid decarboxylase levels in the mouse. Behav Brain Res 314:190–198
Williams G, King J, Cunningham M, Stephan M, Kerr B, Hersh JH (2001) Fetal valproate syndrome and autism: additional evidence of an association. Dev Med Child Neurol 43(3):202–206
Wing L (1997) The autistic spectrum. Lancet 350:1761–1766
Win-Shwe TT, Nway NC, Imai M, Lwin TT, Mar O, Watanabe H (2018) Social behavior, neuroimmune markers and glutamic acid decarboxylase levels in a rat model of valproic acid-induced autism. J Toxicol Sci 43(11):631–643
Yip J, Soghomonian JJ, Blatt GJ (2007) Decreased GAD67 mRNA levels in cerebellar Purkinje cells in autism: pathophysiological implications. Acta Neuropathol 113(5):559–568
Zhao H, Wang Q, Yan T et al (2019) Maternal valproic acid exposure leads to neurogenesis defects and autism-like behaviors in non-human primates. Transl Psychiatry 9(1):267
Zieminska E, Toczylowska B, Diamandakis D, Hilgier W, Filipkowski RK, Polowy R, Orzel J, Gorka M, Lazarewicz JW (2018) Glutamate, glutamine and GABA levels in rat brain measured using MRS, HPLC and NMR methods in study of two models of autism. Front Mol Neurosci 11:418
Zimmerman R, Patel R, Smith A, Pasos J, Kulesza RJ (2018) Repeated prenatal exposure to valproic acid results in auditory brainstem hypoplasia and reduced calcium binding protein immunolabeling. Neuroscience 377:53–68
Zimmerman R, Smith A, Fech T, Mansour Y, Kulesza RJ Jr (2020) In utero exposure to valproic acid disrupts ascending projections to the central nucleus of the inferior colliculus from the auditory brainstem. Exp Brain Res 238(3):551–563. https://doi.org/10.1007/s00221-020-05729-7
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Randy Kulesza designed the study. Stereotaxic injections, perfusions, and tissue processing were done by Randy Kulesza and Yusra Mansour. Neuron counts and analysis of neuron morphology were done by Yusra Mansour and Syed Ahmed. The 3D model was made by Yusra Mansour. All authors have contributed to data analysis. The first draft of the manuscript was written by Randy Kulesza, and all authors have contributed to revisions and editing of the manuscript. Randy Kulesza and Yusra Mansour constructed all figures. All authors read and approved the final manuscript.
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Mansour, Y., Ahmed, S.N. & Kulesza, R. Abnormal morphology and subcortical projections to the medial geniculate in an animal model of autism. Exp Brain Res 239, 381–400 (2021). https://doi.org/10.1007/s00221-020-05982-w
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DOI: https://doi.org/10.1007/s00221-020-05982-w