Assessment of glial function in the in vivo retina

Anja I. Srienc, Tess E. Kornfield, Anusha Mishra, Michael A. Burian, Eric A. Newman

Research output: Chapter in Book/Report/Conference proceedingChapter

7 Citations (Scopus)

Abstract

Glial cells, traditionally viewed as passive elements in the CNS, are now known to have many essential functions. Many of these functions have been revealed by work on retinal glial cells. This work has been conducted almost exclusively on ex vivo preparations and it is essential that retinal glial cell functions be characterized in vivo as well. To this end, we describe an in vivo rat preparation to assess the functions of retinal glial cells. The retina of anesthetized, paralyzed rats is viewed with confocal microscopy and laser speckle flowmetry to monitor glial cell responses and retinal blood flow. Retinal glial cells are labeled with the Ca 2+ indicator dye Oregon Green 488 BAPTA-1 and the caged Ca 2+ compound NP-EGTA by injection of the compounds into the vitreous humor. Glial cells are stimulated by photolysis of caged Ca 2+ and the activation state of the cells assessed by monitoring Ca 2+ indicator dye fluorescence. We find that, as in the ex vivo retina, retinal glial cells in vivo generate both spontaneous and evoked intercellular Ca 2+ waves. We also find that stimulation of glial cells leads to the dilation of neighboring retinal arterioles, supporting the hypothesis that glial cells regulate blood flow in the retina. This in vivo preparation holds great promise for assessing glial cell function in the healthy and pathological retina.

Original languageEnglish (US)
Title of host publicationAstrocytes: Methods and Protocols
Pages499-514
Number of pages16
Volume814
DOIs
StatePublished - 2012
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume814
ISSN (Print)10643745

Fingerprint

Neuroglia
Retina
Vitreous Body
Rheology
Photolysis
Arterioles
Confocal Microscopy
Dilatation
Lasers
Fluorescence
Injections

Keywords

  • Astrocyte
  • Blood flow
  • Calcium wave
  • Confocal microscopy
  • Glial cell
  • In vivo preparation
  • Intracellular calcium
  • Laser speckle flowmetry
  • Müller cell
  • Retina

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Srienc, A. I., Kornfield, T. E., Mishra, A., Burian, M. A., & Newman, E. A. (2012). Assessment of glial function in the in vivo retina. In Astrocytes: Methods and Protocols (Vol. 814, pp. 499-514). (Methods in Molecular Biology; Vol. 814). https://doi.org/10.1007/978-1-61779-452-0_33

Assessment of glial function in the in vivo retina. / Srienc, Anja I.; Kornfield, Tess E.; Mishra, Anusha; Burian, Michael A.; Newman, Eric A.

Astrocytes: Methods and Protocols. Vol. 814 2012. p. 499-514 (Methods in Molecular Biology; Vol. 814).

Research output: Chapter in Book/Report/Conference proceedingChapter

Srienc, AI, Kornfield, TE, Mishra, A, Burian, MA & Newman, EA 2012, Assessment of glial function in the in vivo retina. in Astrocytes: Methods and Protocols. vol. 814, Methods in Molecular Biology, vol. 814, pp. 499-514. https://doi.org/10.1007/978-1-61779-452-0_33
Srienc AI, Kornfield TE, Mishra A, Burian MA, Newman EA. Assessment of glial function in the in vivo retina. In Astrocytes: Methods and Protocols. Vol. 814. 2012. p. 499-514. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61779-452-0_33
Srienc, Anja I. ; Kornfield, Tess E. ; Mishra, Anusha ; Burian, Michael A. ; Newman, Eric A. / Assessment of glial function in the in vivo retina. Astrocytes: Methods and Protocols. Vol. 814 2012. pp. 499-514 (Methods in Molecular Biology).
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