Skip to main content
SpringerLink
Log in

Mapping Auditory Synaptic Circuits with Photostimulation of Caged Glutamate

  • Protocol
  • First Online:
Book cover Auditory and Vestibular Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1427))

Abstract

Photostimulation of neurons with caged glutamate is a viable tool for mapping the strength and spatial distribution of synaptic networks in living brain slices. In photostimulation experiments, synaptic connectivity is assessed by eliciting action potentials in putative presynaptic neurons via focal photolysis of caged glutamate, while measuring postsynaptic responses via intracellular recordings. Two approaches are commonly used for delivering light to small, defined areas in the slice preparation; an optical fiber-based method and a laser-scanning-based method. In this chapter, we outline the technical bases for using photostimulation of caged glutamate to map synaptic circuits, and discuss the advantages and disadvantages of using fiber-based vs. laser-based systems.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Callaway EM, Katz LC (1993) Photostimulation using caged glutamate reveals functional circuitry in living brain slices. Proc Natl Acad Sci U S A 90(16):7661–7665

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Kim G, Kandler K (2003) Elimination and strengthening of glycinergic/GABAergic connections during tonotopic map formation. Nat Neurosci 6(3):282–290

    Article  CAS  PubMed  Google Scholar 

  3. Zhao C, Kao JP, Kanold PO (2009) Functional excitatory microcircuits in neonatal cortex connect thalamus and layer 4. J Neurosci 29(49):15479–15488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Shepherd GM (2012) Circuit mapping by UV uncaging of glutamate. Cold Spring Harb Protoc 9:998–1004

    Google Scholar 

  5. Viswanathan S, Bandyopadhyay S, Kao JP, Kanold PO (2012) Changing microcircuits in the subplate of the developing cortex. J Neurosci 32(5):1589–1601

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Kandler K, Nguyen T, Noh J, Givens RS (2013) An optical fiber-based uncaging system. Cold Spring Harb Protoc 2:118–121

    Google Scholar 

  7. Hirtz JJ, Braun N, Griesemer D, Hannes C, Janz K, Löhrke S, Müller B, Friauf E (2012) Synaptic refinement of an inhibitory topographic map in the auditory brainstem requires functional CaV1.3 calcium channels. J Neurosci 32(42):14602–14616

    Article  CAS  PubMed  Google Scholar 

  8. Clause A, Kim G, Sonntag M, Weisz CJ, Vetter DE, Rübsamen R, Kandler K (2014) The precise temporal pattern of prehearing spontaneous activity is necessary for tonotopic map refinement. Neuron 82(4):822–835

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Campagnola L, Manis PB (2014) A map of functional synaptic connectivity in the mouse anteroventral cochlear nucleus. J Neurosci 34(6):2214–2230

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Sturm J, Nguyen T, Kandler K (2014) Development of intrinsic connectivity in the central nucleus of the mouse inferior colliculus. J Neurosci 34(45):15032–15046

    Article  PubMed  PubMed Central  Google Scholar 

  11. Barbour DL, Callaway EM (2008) Excitatory local connections of superficial neurons in rat auditory cortex. J Neurosci 28(44):11174–11185

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Lee CC, Imaizumi K (2013) Functional convergence of thalamic and intrinsic projections to cortical layers 4 and 6. Neurophysiology 45(5–6):396–406

    Article  PubMed  PubMed Central  Google Scholar 

  13. Meng X, Kao JPY, Kanold PO (2014) Differential signaling to subplate neurons by spatially specific silent synapses in developing auditory cortex. J Neurosci 34(26):8855–8864

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Watkins PV, Kao JPY, Kanold PO (2014) Spatial pattern of intra-laminar connectivity in supragranular mouse auditory cortex. Front Neural Circ 8:15

    Google Scholar 

  15. Kratz MB, Manis PB (2015) Spatial organization of excitatory synaptic inputs to layer 4 neurons in mouse primary auditory cortex. Front Neural Circ 9:17

    Google Scholar 

  16. Stensrud K, Noh J, Kandler K, Wirz J, Heger D, Givens RS (2009) Competing pathways in the photo-Favorskii rearrangement and release of esters: studies on fluorinated p-hydroxyphenacyl-caged GABA and glutamate phototriggers. J Org Chem 74(15):5219–5227

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Conrad PG 2nd, Givens RS, Weber JF, Kandler K (2000) New phototriggers: extending the p-hydroxyphenacyl pi-pi absorption range. Org Lett 2(11):1545–1547

    Article  CAS  PubMed  Google Scholar 

  18. Givens RS, Rubina M, Wirz J (2012) Applications of p-hydroxyphenacyl (pHP) and coumarin-4-ylmethyl photoremovable protecting groups. Photochem Photobiol Sci 11(3):472–488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Shembekar VR, Chen Y, Carpenter BK, Hess GP (2005) A protecting group for carboxylic acids that can be photolyzed by visible light. Biochemistry 44(19):7107–7114

    Article  CAS  PubMed  Google Scholar 

  20. Rial Verde EM, Zayat L, Etchenique R, Yuste R (2008) Photorelease of GABA with visible light using an inorganic caging group. Front Neural Circ 2:2

    Google Scholar 

  21. Fino E, Araya R, Peterka DS, Salierno M, Etchenique R, Yuste R (2009) RuBi-glutamate: two-photon and visible-light photoactivation of neurons and dendritic spines. Front Neural Circ 3:2

    Google Scholar 

  22. Leszkiewicz DN, Kandler K, Aizenman E (2000) Enhancement of NMDA receptor-mediated currents by light in rat neurones in vitro. J Physiol 524(Pt 2):365–374

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Lesziewicz DN, Aizenman E (2003) Reversible modulation of GABA(A) receptor-mediated currents by light is dependent on the redox state of the receptor. Eur J Neurosci 17(10):2077–2083

    Article  Google Scholar 

  24. Kandler K, Katz LC, Kauer JA (1998) Focal photolysis of caged glutamate reveals an entirely postsynaptic form of long-term depression of hippocampal glutamate receptors. Nat Neurosci 1:119–123

    Article  CAS  PubMed  Google Scholar 

  25. Noh J, Seal RP, Garver JA, Edwards RH, Kandler K (2010) Glutamate co-release at GABA/glycinergic synapses is crucial for refinement of an inhibitory map. Nat Neurosci 13(2):232–238

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Matsuzaki M, Ellis-Davis GC, Nemoto T, Miyashita Y, Lino M, Kasai H (2001) Dendritic spine geometry is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurons. Nat Neurosci 4(11):1086–1092

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Smith MA, Ellis-Davis GC, Magee JC (2003) Mechanism of the distance-dependent scaling of Schaffer collateral synapses in rat CA1 pyramidal neurons. J Physiol 548(Pt. 1):245–258

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Otolaryngology, School of Medicine, Ear and Eye Institute, University of Pittsburgh, 3501 Fifth Avenue, Pittsburgh, PA, 15213, USA

    Joshua J. Sturm & Karl Kandler

  2. Department of Neurobiology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Joshua J. Sturm & Karl Kandler

  3. Medical Scientist Training Program, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15261, USA

    Joshua J. Sturm

  4. Department of Physics, The College of New Jersey, Ewing, NJ, 08628, USA

    Tuan Nguyen

  5. Center for the Neural Basis of Cognition, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Karl Kandler

Authors
  1. Joshua J. Sturm
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tuan Nguyen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Karl Kandler
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Karl Kandler .

Editor information

Editors and Affiliations

  1. College of Medicine, University of South Florida, Tampa, Florida, USA

    Bernd Sokolowski

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Sturm, J.J., Nguyen, T., Kandler, K. (2016). Mapping Auditory Synaptic Circuits with Photostimulation of Caged Glutamate. In: Sokolowski, B. (eds) Auditory and Vestibular Research. Methods in Molecular Biology, vol 1427. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3615-1_30

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3615-1_30

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3613-7

  • Online ISBN: 978-1-4939-3615-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation