Correlative light and electron microscopic observation of calcium phosphate particles in a mouse kidney formed under a high-phosphate diet

Calcium phosphate forms particles under excessive urinary excretion of phosphate in the kidney. While the formation of calcium phosphate particles (CaPs) has been implicated in the damage to renal tubular cells and renal dysfunction, clarifying the ultrastructural information and the elemental compo...

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Bibliographic Details
Published in:Scientific reports 2023-01, Vol.13 (1), p.852-852, Article 852
Main Authors: Battulga, Batpurev, Shiizaki, Kazuhiro, Miura, Yutaka, Osanai, Yasuyuki, Yamazaki, Reiji, Shinohara, Yoshiaki, Kubota, Yoshiyuki, Hara, Toru, Kuro-o, Makoto, Ohno, Nobuhiko
Format: Article
Language:English
Subjects:
631/1647/328/1649
692/4019/592/2193
692/4022/1585/1732
Alendronic acid
Animals
Bisphosphonates
Calcium Phosphates
Chemical composition
Diet
Electrons
Fluorescent indicators
Freezing
Humanities and Social Sciences
Kidney
Kidneys
Magnesium
Mice
Microscopy
Microscopy, Electron, Scanning
multidisciplinary
Phosphate
Phosphates
Renal function
Scanning electron microscopy
Science
Science (multidisciplinary)
Toluidine
X-ray spectroscopy
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Summary:Calcium phosphate forms particles under excessive urinary excretion of phosphate in the kidney. While the formation of calcium phosphate particles (CaPs) has been implicated in the damage to renal tubular cells and renal dysfunction, clarifying the ultrastructural information and the elemental composition of the small CaPs in the wide areas of kidney tissue has been technically difficult. This study introduces correlative and sequential light as well as electron microscopic CaP observation in the kidney tissue by combining fluorescent staining for CaPs and energy-dispersive X-ray spectroscopy (EDS) in scanning electron microscopy (SEM) on resin sections prepared using high-pressure freezing and freeze substitution. CaPs formed in mouse kidneys under long-term feeding of a high-phosphate diet were clearly visualized on resin sections by fluorescence-conjugated alendronate derivatives and toluidine blue metachromasia. These CaPs were verified by correlative observation with EDS. Furthermore, small CaPs formed in the kidney under short-term feeding were detected using fluorescent probes. The elemental composition of the particles, including calcium and magnesium, was identified following EDS analyses. These results suggest that the correlative microscopy approach is helpful for observing in situ distribution and elemental composition of CaPs in the kidney and contributing to studies regarding CaP formation-associated pathophysiology.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-023-28103-3