June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Astrocyte morphological differences between collagenous and glial lamina
Author Affiliations & Notes
  • Ian A Sigal
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
    Bioengineering, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Susannah Waxman
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Footnotes
    Commercial Relationships   Ian Sigal None; Susannah Waxman None
  • Footnotes
    Support  Supported in part by National Institutes of Health grants R01-EY023966, R01-EY031708, R01-HD045590, R01-HD083383, 1S10RR028478-01, P30-EY008098, and T32-EY017271; Eye and Ear Foundation (Pittsburgh, PA); Research to Prevent Blindness (unrestricted grant to UPMC Ophthalmology and Stein Innovation Award to Sigal IA).
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 3782. doi:
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    • Get Citation

      Ian A Sigal, Susannah Waxman; Astrocyte morphological differences between collagenous and glial lamina. Invest. Ophthalmol. Vis. Sci. 2023;64(8):3782.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Much of what is known about individual astrocyte morphology in the lamina is based on rodents. Rodents, however, have a glial lamina (GL) without the collagenous beams existing in primates. Our goal was to quantify individual astrocyte morphology of the monkey collagenous lamina cribrosa (CL) and test the hypothesis that astrocyte morphology is different between CL and GL

Methods : Coronal vibratome sections were obtained through the CL of 7 monkey eyes at 150µm thickness. Gold microcarriers coated in cell membrane dyes were ballistically delivered into sections for Multicolor DiOlistic labeling. Confocal microscopy and second harmonic generation (SHG) imaging were used to visualize dyed astrocytes and collagen beams, respectively. 3D models of 28 dyed CL astrocytes were constructed for automated quantification of morphological features. GL astrocyte morphological features were collected from the literature

Results : Mean ± SD CL astrocyte branch number, length, thickness, hierarchy, and straightness were 86.5 ± 45.7, 14.0 ± 13.5µm, 2.1 ± 1.4µm, 6.1 ± 3.7, and 0.9 ± 0.1. Sholl analysis of CL astrocyte models revealed higher branching complexity than mouse GL astrocytes, indicated by area under Sholl curves (p < 0.001). Diameter of CL first and second order astrocyte branches was 3.1± 0.15µm (mean ± SEM), significantly thicker than comparable GL astrocyte branches (1.3 ± 0.04μm, p < 0.001). SHG revealed CL astrocytes encircling beams and spanning multiple pores

Conclusions : CL astrocytes display significant morphological differences from those in the GL. Greater branching complexity in CL astrocytes supports potentially more complex interactions of these astrocytes with retinal ganglion cell axons and blood vessels, resulting in possible functional differences. Low-order branch thickness in healthy CL astrocytes was more similar to that of the thickened astrocyte branches in mouse glaucoma models than healthy mouse controls. The ability of CL astrocytes to span multiple pores and interact with CL beams may result in differences in signaling compared to the GL, in which all astrocytes exist within a single pore-like canal. The implications of these differences should be investigated to better understand astrocyte physiology in the CL and their role in health and disease

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

 

A) Sholl analysis and B) low-order branch thickness of CL vs GL astrocytes. C) Sholl analysis and D) branch thickness of example models. Error bars: SEM.

A) Sholl analysis and B) low-order branch thickness of CL vs GL astrocytes. C) Sholl analysis and D) branch thickness of example models. Error bars: SEM.

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