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Molecular, anatomical and functional changes in the retinal ganglion cells after optic nerve crush in mice

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Abstract

Purpose

Optic nerve crush (ONC) and subsequent axonal damage can be used in rodents to study the mechanism of retinal ganglion cell (RGC) degeneration. Here, we examined electroretinograms (ERGs) in post-ONC mice to investigate changes in the positive scotopic threshold response (pSTR). We then compared these changes with molecular and morphological changes to identify early objective biomarkers of RGC dysfunction.

Methods

Fifty 12-week-old C57BL/6 mice were included. ONC was used to induce axonal injury in the right eye of each animal, with the left eye used as a control. The expression of the RGC markers Brn3a and Brn3b was measured on days 1, 2, 3, 5 and 7 after ONC with quantitative real-time PCR. ERGs were recorded under dark adaptation with the stimulus intensity increasing from −6.2 to 0.43 log cd-s/m2 on days 1, 2, 3, 5, 7 and 10 after ONC. The pSTR, a- and b-wave amplitudes were measured. Inner retinal thickness around the optic nerve head was measured with spectral-domain optical coherence tomography on days 0, 2, 5, 7 and 10 after ONC.

Results

The expression of Brn3a and Brn3b began to significantly decrease on day 1 and day 2, respectively (P < 0.01). The amplitude of the pSTR underwent rapid, significant deterioration on day 3, after which it fell gradually (P < 0.01), while the a- and b-wave amplitudes remained unchanged throughout the experiment. Inner retinal thickness gradually decreased, with the most significant reduction on day 10 (P < 0.01).

Conclusions

Decrease in pSTR likely reflected the early loss of RGC function after ONC and that declining expression of RGC-specific genes preceded anatomical and functional changes in the RGCs.

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Acknowledgments

We thank Ms. Junko Sato for her technical assistance. This work was supported in part by JSPS KAKENHI Grants-in-Aid for Scientific Research B (T.N. 26293372) and for Challenging Exploratory Research (Y.T. 26670263 and T.N. 26670751). This study was also supported by the JST Center for Revitalization Promotion (Y.T. and T.N.).

Conflict of interest

The authors have no conflict of interest.

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Authors and Affiliations

  1. Department of Ophthalmology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, Miyagi, 980-8574, Japan

    Masayoshi Yukita, Satoru Tsuda, Yu Yokoyama, Masayuki Yasuda, Kazuichi Maruyama & Toru Nakazawa

  2. Department of Retinal Disease Control, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan

    Toru Nakazawa

  3. Department of Advanced Ophthalmic Medicine, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan

    Koji M. Nishiguchi & Toru Nakazawa

  4. Department of Ophthalmology, Dokkyo Medical University Koshigaya Hospital, Saitama, Japan

    Shigeki Machida

  5. Department of Ophthalmology, Iwate Medical University School of Medicine, Morioka, Japan

    Shigeki Machida

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  1. Masayoshi Yukita
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  2. Shigeki Machida
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  3. Koji M. Nishiguchi
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  4. Satoru Tsuda
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  5. Yu Yokoyama
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  7. Kazuichi Maruyama
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  8. Toru Nakazawa
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Correspondence to Toru Nakazawa.

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Yukita, M., Machida, S., Nishiguchi, K.M. et al. Molecular, anatomical and functional changes in the retinal ganglion cells after optic nerve crush in mice. Doc Ophthalmol 130, 149–156 (2015). https://doi.org/10.1007/s10633-014-9478-2

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  • DOI: https://doi.org/10.1007/s10633-014-9478-2

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