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Ultrastructural changes in the choriocapillaris of N-methyl-N-nitrosourea-induced retinal degeneration in C57BL/6 mice

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Abstract

N-methyl-N-nitrosourea (MNU) is known to cause apoptosis of photoreceptor cells and changes in retinal pigment epithelium (RPE). However, the changes in choriocapillaris, which nourishes photoreceptor cells by diffusing tissue fluid through RPE, have not been reported in detail. Therefore, we studied the ultrastructural transformation in and around the choriocapillaris to characterize the interdependence between choriocapillaris and surrounding tissue components in a mouse model. Seven-week-old male C57BL/6 mice were given a single intraperitoneal injection of MNU (60 mg/kg of body weight). Perfusion-fixed eyeballs were examined chronologically using immunohistochemistry and electron microscopy until the photoreceptor cells were lost. Sequential ultrastructural changes were observed in photoreceptor cells, RPE, Bruch’s membrane, choriocapillaris, and choroidal melanocytes after an MNU injection. The lumens of the choriocapillaris narrowed following dilation, and the vascular endothelium showed structural alterations. When the photoreceptor cells were completely lost, the choriocapillaris appeared to be in a recovery process. Our results suggest that transport abnormality through Bruch’s membrane and structural changes in the choroid might have influenced the morphology of choriocapillaris. The thin wall of the choriocapillaris appears to be the cause of the vulnerability with its altered morphology.

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Abbreviations

MNU:

N-Methyl-N-nitrosourea

RPE:

Retinal pigment epithelium

CC:

Choriocapillaris

ONL:

Outer nuclear layer

OPL:

Outer plexiform layer

IS:

Inner segments

OS:

Outer segments

GCL:

Ganglion cell layer

IPL:

Inner plexiform layer

INL:

Inner nuclear layer

BL:

Basal lamina

OCL:

Outer collagenous layer

ICL:

Inner collagenous layer

PB:

Phosphate buffer

PBS:

Phosphate-buffered saline

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Acknowledgements

We thank Ms. Sachie Matsubara and Ms. Junri Hayakawa of the Laboratory for Electron Microscopy, Kyorin University School of Medicine, who employed their expertise in helping our team to obtain excellent digital electron microscopic images for this research. This study was initiated by Shunichi Morikawa (Ph.D.), a devoted researcher at Tokyo Woman’s Medical University. His passion to investigate the choroidal microcirculation was diligently inherited by our team after his untimely death at a young age in 2017. We are indebted to him for his dedication and leadership in making this research happen. We are also thankful to Ms. Hiromi Sagawa and Ms. Kae Motomaru, Department of Microscopic and Developmental Anatomy, Tokyo Women’s Medical University, for constantly supporting us to achieve this goal. Finally, we would like to thank Ms. Kaoru Otsuka for language support in producing the English article.

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Authors and Affiliations

  1. Department of Anatomy and Developmental Biology, School of Medicine, Tokyo Women’s Medical University, 8-1 Kawada-cho, Shinjuku, Tokyo, 162-8666, Japan

    Ruriko Hayakawa, Masae Morishima, Shuji Kitahara & Taichi Ezaki

  2. Department of Anatomy, School of Medicine, Tokyo Women’s Medical University, Shinjuku, Tokyo, 162-8666, Japan

    Kaori Komoike & Hiroki Fujieda

  3. Department of Anatomy, Kyorin University School of Medicine, Mitaka, Tokyo, 181-8611, Japan

    Hayato Kawakami

  4. Department of Anatomy, School of Medicine, Toho University, Ota, Tokyo, 143-8540, Japan

    Kazuhiko Shimizu

  5. Department of Ophthalmology, Kyorin University School of Medicine, Mitaka, Tokyo, 181-8611, Japan

    Ruriko Hayakawa

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  1. Ruriko Hayakawa
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  2. Kaori Komoike
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  3. Hayato Kawakami
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Correspondence to Ruriko Hayakawa.

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Hayakawa, R., Komoike, K., Kawakami, H. et al. Ultrastructural changes in the choriocapillaris of N-methyl-N-nitrosourea-induced retinal degeneration in C57BL/6 mice. Med Mol Morphol 53, 198–209 (2020). https://doi.org/10.1007/s00795-020-00246-6

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