Top

Journal of the Association for Research in Otolaryngology

Published in:

30-05-2023 | Ultrasound | Original Article

Connexin36 RNA Expression in the Cochlear Nucleus of the Echolocating Bat, Eptesicus fuscus

Authors: Alyssa W. Accomando, Mark A. Johnson, Madeline A. McLaughlin, James A. Simmons, Andrea Megela Simmons

Published in: Journal of the Association for Research in Otolaryngology | Issue 3/2023

Abstract

Purpose

The echolocating bat is used as a model for studying the auditory nervous system because its specialized sensory capabilities arise from general mammalian auditory percepts such as pitch and sound source localization. These percepts are mediated by precise timing within neurons and networks of the lower auditory brainstem, where the gap junction protein Connexin36 (CX36) is expressed. Gap junctions and electrical synapses in the central nervous system are associated with fast transmission and synchronous patterns of firing within neuronal networks. The purpose of this study was to identify areas where CX36 was expressed in the bat cochlear nucleus to shed light on auditory brainstem networks in a hearing specialist animal model.

Methods

We investigated the distribution of CX36 RNA throughout the cochlear nucleus complex of the echolocating big brown bat, Eptesicus fuscus, using in situ hybridization. As a qualitative comparison, we visualized Gjd2 gene expression in the cochlear nucleus of transgenic CX36 reporter mice, species that hear ultrasound but do not echolocate.

Results

In both the bat and the mouse, CX36 is expressed in the anteroventral and in the dorsal cochlear nucleus, with more limited expression in the posteroventral cochlear nucleus. These results are generally consistent with previous work based on immunohistochemistry.

Conclusion

Our data suggest that the anatomical substrate for CX36-mediated electrical neurotransmission is conserved in the mammalian CN across echolocating bats and non-echolocating mice.

Bennett MV, Zukin RS (2004) Electrical coupling and neuronal synchronization in the mammalian brain. Neuron 41(4):495–511. https://doi.org/10.1016/S0896-6273(04)00043-1CrossRefPubMed

Connors BW (2017) Synchrony and so much more: diverse roles for electrical synapses in neural circuits. Dev Neurobiol 77(5):610–624. https://doi.org/10.1002/dneu.22493CrossRefPubMedPubMedCentral

Pereda AE (2014) Electrical synapses and their functional interactions with chemical synapses. Nat Rev Neurosci 15(4):250–263. https://doi.org/10.1038/nrn3708CrossRefPubMedPubMedCentral

Galarreta M, Hestrin S (2001) Spike transmission and synchrony detection in networks of GABAergic interneurons. Science 292(5525):2295–2299. https://doi.org/10.1126/science.1061395CrossRefPubMed

Joris PX, Smith PH, Yin TC (1998) Coincidence detection in the auditory system: 50 years after Jeffress. Neuron 21(6):1235–1238. https://doi.org/10.1016/S0896-6273(00)80643-1CrossRefPubMed

Licklider JCR (1959) Three auditory theories. In: Koch S (ed) Psychology: a study of a science. McGraw-Hill, New York, pp 41–144

Simmons JA, Simmons AM (2011) Bats and frogs and animals in between: evidence for a common central timing mechanism to extract periodicity pitch. J Comp Physiol A Neuroethol Sens Neural Behav Physiol 197(5):585–594. https://doi.org/10.1007/s00359-010-0607-4CrossRefPubMed

Lyon RF (2017) Human and machine hearing. Cambridge University Press, Cambridge UKCrossRef

Ming C, Haro S, Simmons AM, Simmons JA (2021) A comprehensive computational model of animal biosonar signal processing. PLoS Comput Biol 17(2):e1008677. https://doi.org/10.1371/journal.pcbi.1008677

10.

Rash JE, Staines WA, Yasumura T, Patel D, Furman CS, Stelmack GL, Nagy JI (2000) Immunogold evidence that neuronal gap junctions in adult rat brain and spinal cord contain connexin-36 but not connexin-32 or connexin-43. Proc Natl Acad Sci USA 97(13):7573–7578. https://doi.org/10.1073/pnas.97.13.7573CrossRefPubMedPubMedCentral

11.

Rubio ME, Nagy JI (2015) Connexin36 expression in major centers of the auditory system in the CNS of mouse and rat: evidence for neurons forming purely electrical synapses and morphologically mixed synapses. Neurosci 303:604–629. https://doi.org/10.1016/j.neuroscience.2015.07.026CrossRef

12.

Belluardo N, Mudo G, Trovato-Salinaro A, Le Gurun S, Charollais A, Serre-Beinier V, Amato G, Haefliger JA, Meda P, Condorelli DF (2000) Expression of connexin36 in the adult and developing rat brain. Brain Res 865(1):121–138. https://doi.org/10.1016/S0006-8993(00)02300-3CrossRefPubMed

13.

Condorelli DF, Parenti R, Spinella F, Trovato Salinaro A, Belluardo N, Cardile V, Cicirata F (1998) Cloning of a new gap junction gene (Cx36) highly expressed in mammalian brain neurons. Eur J Neurosci 10(3):1202–1208. https://doi.org/10.1046/j.1460-9568.1998.00163.xCrossRefPubMed

14.

Gómez-Nieto R, Rubio ME (2009) A bushy cell network in the rat ventral cochlear nucleus. J Comp Neurol 516(4):241–263. https://doi.org/10.1002/cne.22139CrossRefPubMedPubMedCentral

15.

Mugnaini E (1985) GABA neurons in the superficial layers of the rat dorsal cochlear nucleus: light and electron microscopic immunocytochemistry. J Comp Neurol 235(1):61–81. https://doi.org/10.1002/cne.902350106CrossRefPubMed

16.

Sotelo CT, Gentschev T, Zamora AJ (1976) Gap junctions in ventral cochlear nucleus of the rat. A possible new example of electrotonic junctions in the mammalian C.N.S. Neurosci 1(1):5–7. https://doi.org/10.1016/0306-4522(76)90041-5

17.

Wouterlood FG, Mugnaini E, Osen KK, Dahl AL (1984) Stellate neurons in rat dorsal cochlear nucleus studies with combined Golgi impregnation and electron microscopy: synaptic connections and mutual coupling by gap junctions. J Neurocytol 13(4):639–664. https://doi.org/10.1007/BF01148083CrossRefPubMed

18.

Gómez-Nieto R, Rubio ME (2011) Ultrastructure, synaptic organization, and molecular components of bushy cell networks in the anteroventral cochlear nucleus of the rhesus monkey. Neurosci 179:188–207. https://doi.org/10.1016/j.neuroscience.2011.01.058CrossRef

19.

Apostolides PF, Trussell LO (2013) Regulation of interneuron excitability by gap junction coupling with principal cells. Nat Neurosci 16(12):1764–1772. https://doi.org/10.1038/nn.3569CrossRefPubMedPubMedCentral

20.

Apostolides PF, Trussell LO (2014a) Control of interneuron firing by subthreshold synaptic potentials in principal cells of the dorsal cochlear nucleus. Neuron 83(2):324–330. https://doi.org/10.1016/j.neuron.2014.06.008CrossRefPubMedPubMedCentral

21.

Apostolides PF, Trussell LO (2014b) Superficial stellate cells of the dorsal cochlear nucleus. Front Neural Circuits 8:63. https://doi.org/10.3389/fncir.2014.00063CrossRefPubMedPubMedCentral

22.

Yaeger DB, Trussell LO (2016) Auditory Golgi cells are interconnected predominantly by electrical synapses. J Neurophysiol 116(2):540–551. https://doi.org/10.1152/jn.01108.2015CrossRefPubMedPubMedCentral

23.

Cant NB (1992) The cochlear nucleus: neuronal types and their synaptic organization. In: DB Webster, AN Popper, RR Fay (eds), The Mammalian Auditory Pathway: Neuroanatomy. Springer Handbook of Auditory Research, vol 1. Springer, New York, pp 66–116. https://doi.org/10.1007/978-1-4612-4416-5_3

24.

Rubio ME (2018) Microcircuits of the ventral cochlear nucleus. In: DL Oliver, NB Cant, RR Fay, AN Popper (eds), The mammalian auditory pathways: synaptic organization and microcircuits. Springer Handbook of Auditory Research, vol 65. Springer, Cham, pp 41–71. https://doi.org/10.1007/978-3-319-71798-2_3

25.

Trussell LO, Oertel D (2018) Microcircuits of the dorsal cochlear nucleus. In: DL Oliver, NB Cant, RR Fay, AN Popper (eds), The mammalian auditory pathways: synaptic organization and microcircuits. Springer Handbook of Auditory Research, vol 65. Springer, Cham, pp 73–99. https://doi.org/10.1007/978-3-319-71798-2_4

26.

Joris PX, Carney LH, Smith PH, Yin TC (1994) Enhancement of neural synchronization in the anteroventral cochlear nucleus. I. Responses to tones at the characteristic frequency. J Neurophysiol 71(3):1022–1036. https://doi.org/10.1152/jn.1994.71.3.1022

27.

Horowitz SS, Stamper SA, Simmons JA (2008) Neuronal connexin expression in the cochlear nucleus of big brown bats. Brain Res 1197:76–84. https://doi.org/10.1016/j.brainres.2007.12.048CrossRefPubMed

28.

Covey E, Casseday JH (1999) Timing in the auditory system of the bat. Ann Rev Physiol 61:457–476. https://doi.org/10.1146/annurev.physiol.61.1.457CrossRef

29.

Luo J, Maciás S, Ness TV, Einevoll GT, Zhang K, Moss CF (2018) Neural timing of stimulus events with microsecond precision. PLOS Biol 16(10):e2006422. https://doi.org/10.1371/journal.pbio.2006422

30.

Simmons JA (1993) Evidence for perception of fine echo delay and phase by the FM bat, Eptesicus fuscus. J Comp Physiol A 172:533–547. https://doi.org/10.1007/BF00213677CrossRefPubMed

31.

Simmons JA, Ferragamo MJ, Sanderson MI (2003) Echo delay versus spectral cues for temporal hyperacuity in the big brown bat, Eptesicus fuscus. J Comp Physiol Neuroethol Sens Neural Behav Physiol 189:693–702. https://doi.org/10.1007/s00359-003-0444-9CrossRef

32.

Di Palma F, Alfoldi J, Johnson J, Berlin A, Gnerre S, MacCallum JDI, Young S, Walker BJ, Lindblad-Toh K (2012) The draft genome of Eptesicus fuscus. The Broad Institute Genome Assembly & Analysis Group, Computational R&D Group, and Sequencing Platform. https://www.ncbi.nlm.nih.gov/assembly/GCF_000308155.1/

33.

Timothy M, Forlano PM (2019) A versatile macro-based neurohistological images analysis suite for ImageJ focused on automated and standardized user interaction and reproducible data output. J Neurosci Meth 324(108286). https://doi.org/10.1016/j.jneumeth.2019.04.009

34.

Rasband WS (1997–2018) ImageJ. U. S. National Institutes of Health, Bethesda, Maryland, USA. https://imagej.nih.gov/ij/, 1997–2018

35.

Deans MR, Gibson JR, Sellitto C, Connors BW, Paul DL (2001) Synchronous activity of inhibitory networks in neocortex requires electrical synapses containing connexin36. Neuron 31(3):477–485. https://doi.org/10.1016/S0896-6273(01)00373-7CrossRefPubMed

36.

Covey E, Casseday JH (1995) The lower brainstem auditory pathways. In: AN Popper, RR Fay (eds), Hearing by bats. Springer Handbook of Auditory Research, vol 5. Springer, New York, pp 235–295. https://doi.org/10.1007/978-1-4612-2556-0_6

37.

Hall JG (1969) The cochlea and the cochlear nuclei in the bat. Acta Otolaryngol 67(5):490–500. https://doi.org/10.3109/00016486909125476CrossRefPubMed

38.

Haplea S, Covey E, Casseday JH (1994) Frequency tuning and response latencies at three levels in the brainstem of the echolocating bat, Eptesicus fuscus. J Comp Physiol A Sens Neural Behav Physiol 174(6):671–683. https://doi.org/10.1007/BF00192716CrossRef

39.

Rosenberger MH, Fremouw T, Casseday JH, Covey E (2003) Expression of the Kv1.1 ion channel subunit in the auditory brainstem of the big brown bat, Eptesicus fuscus. J Comp Neurol 462(1):101–120. https://doi.org/10.1002/cne.10713

40.

Godfrey DA, Lee AC, Hamilton WD, Benjamin LC, Vishwanath S, Simo H, Godfrey LM, Mustapha AIAA, Heffner RS (2016) Volumes of cochlear nucleus regions in rodents. Hear Res 339:161–174. https://doi.org/10.1016/j.heares.2016.07.003CrossRefPubMedPubMedCentral

41.

Vater M (1982) Single unit responses in cochlear nucleus of horseshoe bats to sinusoidal frequency and amplitude modulated signals. J Comp Physiol 149:369–388. https://doi.org/10.1007/BF00619153CrossRef

42.

Lauer AM, Connelly CJ, Graham H, Ryugo DK (2013) Morphological characterization of bushy cells and their inputs in the laboratory mouse (Mus musculus) anteroventral cochlear nucleus. PLoS ONE 8(8):e73308. https://doi.org/10.1371/journal.pone.0073308

43.

Oertel D, Bal R, Gardner SM, Smith PH, Joris PX (2000) Detection of synchrony in the activity of auditory nerve fibers by octopus cells of the mammalian cochlear nucleus. Proc Natl Acad Sci USA 97:11773–11779. https://doi.org/10.1073/pnas.97.22.11773CrossRefPubMedPubMedCentral

44.

Kemmer M, Vater M (1997) The distribution of GABA and glycine immunostaining in the cochlear nucleus of the mustached bat (Pteronotus parnellii). Cell Tissue Res 287:487–506. https://doi.org/10.1007/s004410050773CrossRefPubMed

45.

Rubio ME, Gudsnuk KA, Smith Y, Ryugo DK (2008) Revealing the molecular layer of the primate dorsal cochlear nucleus. Neurosci 154:99–113. https://doi.org/10.1016/j.neuroscience.2007.12.016CrossRef

46.

Carter ME (2004) A stereotaxic brain atlas of the big brown bat. BatLab, University of Washington, Seattle WA, Eptesicus fuscus

47.

Parenti R, Gulisano M, Zappala’ A, Cicirata F (2000) Expression of connexin36 mRNA in adult rodent brain. NeuroReport 11(7):1497–1502CrossRefPubMed

Title: Connexin36 RNA Expression in the Cochlear Nucleus of the Echolocating Bat, Eptesicus fuscus
Authors: Alyssa W. Accomando
Mark A. Johnson
Madeline A. McLaughlin
James A. Simmons
Andrea Megela Simmons
Publication date: 30-05-2023
Publisher: Springer US
Keywords: Ultrasound
Ultrasound
Published in: Journal of the Association for Research in Otolaryngology / Issue 3/2023
Print ISSN: 1525-3961
Electronic ISSN: 1438-7573
DOI: https://doi.org/10.1007/s10162-023-00898-y

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Connexin36 RNA Expression in the Cochlear Nucleus of the Echolocating Bat, Eptesicus fuscus

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 3/2023

In Vivo Optical Characterization of Middle Ear Effusions and Biofilms During Otitis Media

Nuclear Translocation Triggered at the Onset of Hearing in Cochlear Inner Hair Cells of Rats and Mice

Types of Inheritance and Genes Associated with Familial Meniere Disease

NIDCD’s 5-Year Strategic Plan Describes Scientific Priorities and Commitment to Basic Science

Finite-Element Modelling Based on Optical Coherence Tomography and Corresponding X-ray MicroCT Data for Three Human Middle Ears

Bandpass Shape of Distortion-Product Otoacoustic Emission Ratio Functions Reflects Cochlear Frequency Tuning in Normal-Hearing Mice