title: Differential maturation of vesicular glutamate and GABA transporter expression in the mouse auditory forebrain during the first weeks of hearing.
abstract: Vesicular transporter proteins are an essential component of the presynaptic machinery that regulates neurotransmitter storage and release. They also provide a key point of control for homeostatic signaling pathways that maintain balanced excitation and inhibition following changes in activity levels, including the onset of sensory experience. To advance understanding of their roles in the developing auditory forebrain, we tracked the expression of the vesicular transporters of glutamate (VGluT1, VGluT2) and GABA (VGAT) in primary auditory cortex (A1) and medial geniculate body (MGB) of developing mice (P7, P11, P14, P21, adult) before and after ear canal opening (~P11-P13). RNA sequencing, in situ hybridization, and immunohistochemistry were combined to track changes in transporter expression and document regional patterns of transcript and protein localization. Overall, vesicular transporter expression changed the most between P7 and P21. The expression patterns and maturational trajectories of each marker varied by brain region, cortical layer, and MGB subdivision. VGluT1 expression was highest in A1, moderate in MGB, and increased with age in both regions. VGluT2 mRNA levels were low in A1 at all ages, but high in MGB, where adult levels were reached by P14. VGluT2 immunoreactivity was prominent in both regions. VGluT1 (+) and VGluT2 (+) transcripts were co-expressed in MGB and A1 somata, but co-localization of immunoreactive puncta was not detected. In A1, VGAT mRNA levels were relatively stable from P7 to adult, while immunoreactivity increased steadily. VGAT (+) transcripts were rare in MGB neurons, whereas VGAT immunoreactivity was robust at all ages. Morphological changes in immunoreactive puncta were found in two regions after ear canal opening. In the ventral MGB, a decrease in VGluT2 puncta density was accompanied by an increase in puncta size. In A1, perisomatic VGAT and VGluT1 terminals became prominent around the neuronal somata. Overall, the observed changes in gene and protein expression, regional architecture, and morphology relate to-and to some extent may enable-the emergence of mature sound-evoked activity patterns. In that regard, the findings of this study expand our understanding of the presynaptic mechanisms that regulate critical period formation associated with experience-dependent refinement of sound processing in auditory forebrain circuits.
authors.affiliation: Department of Hearing and Speech Sciences, Vanderbilt University School of Medicine, 465 21st Avenue South, MRB-3 Suite 7110, Nashville, TN, 37232, USA. troy.a.hackett@vanderbilt.edu.
authors.forename: Troy A
authors.initials: TA
authors.lastname: Hackett
authors.affiliation: Eaton-Peabody Laboratories, Massachusetts Eye and Ear Infirmary, Department of Otology and Laryngology, Harvard Medical School, Boston, MA, USA.
authors.forename: Amanda R
authors.initials: AR
authors.lastname: Clause
authors.affiliation: Department of Hearing and Speech Sciences, Vanderbilt University School of Medicine, 465 21st Avenue South, MRB-3 Suite 7110, Nashville, TN, 37232, USA.
authors.forename: Toru
authors.initials: T
authors.lastname: Takahata
authors.affiliation: Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Hackett
authors.affiliation: Eaton-Peabody Laboratories, Massachusetts Eye and Ear Infirmary, Department of Otology and Laryngology, Harvard Medical School, Boston, MA, USA.
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Polley
mesh_terms: D000818:Animals; D001303:Auditory Cortex / Q000254:growth & development / Q000378:metabolism*; D005260:Female; D005829:Geniculate Bodies / Q000254:growth & development / Q000378:metabolism*; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D012333:RNA, Messenger / Q000378:metabolism; D017423:Sequence Analysis, RNA; D050598:Vesicular Glutamate Transport Protein 1 / Q000235:genetics / Q000378:metabolism*; D050599:Vesicular Glutamate Transport Protein 2 / Q000235:genetics / Q000378:metabolism*; D050495:Vesicular Inhibitory Amino Acid Transport Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D012333:RNA, Messenger; C493477:Slc17a6 protein, mouse; C493465:Slc17a7 protein, mouse; D050598:Vesicular Glutamate Transport Protein 1; D050599:Vesicular Glutamate Transport Protein 2; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; C492244:Viaat protein, mouse
keywords: A1; BDNF; Cortex; Critical period; Development; Fox-3; GABA; Geniculate; Glutamate; Homeostatic plasticity; Juvenile; MeCP2; Protein; RNA; Sequencing; Thalamus