Morphological Features of the Kidneys in INSRR Knockout Mice Under Bicarbonate Load

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Abstract

BACKGROUND: The insulin receptor-related receptor (IRR), a receptor tyrosine kinase, functions as a sensor of extracellular alkaline pH and is involved in renal bicarbonate excretion. High IRR expression has been detected in β-intercalated cells of the kidney, located in the distal tubules where bicarbonate secretion occurs. To create a new model for investigating sensitivity to pH changes, a unique INSRR knockout mouse line was developed on a C57BL/6 background.

AIM: To analyze morphological changes in kidney tissue in INSRR knockout mice compared with wild-type animals under normal conditions and during metabolic alkalosis.

METHODS: The study used littermate mice from a single generation, with genotypes confirmed by polymerase chain reaction. Two mouse lines were used in the experiment: INSRR knockout and wild-type. The animals were studied under two conditions—baseline and experimentally induced alkalosis. Morphometric analysis was performed on hematoxylin and eosin–stained kidney cryosections. The number of macrophages in kidney tissue was evaluated using immunohistochemical staining.

RESULTS: Morphometric analysis revealed that INSRR gene knockout did not lead to significant pathological alterations in kidney structure. However, significant differences were observed in parenchymal thickness, glomerular area, and collecting duct diameter in sections taken at the level of the renal pelvis. Differences were observed both when comparing the two mouse lines under normal conditions and during experimental alkalosis. Additionally, kidney size in knockout mice was smaller than in wild-type animals. Immunohistochemical analysis revealed no statistically significant differences in the number of CD206-positive (anti-inflammatory) macrophages in the kidneys under either normal conditions or experimental alkalosis.

CONCLUSION: Morphometric analysis of histological sections revealed increased parenchymal thickness in receptor tyrosine kinase knockout mice compared with wild-type animals under experimental alkalosis. Overall, knockout of the IRR receptor tyrosine kinase gene did not result in major pathological changes in kidney architecture. Thus, this genetically modified mouse line may serve as a model for physiological and molecular biological studies of metabolic alkalosis and its associated pathological processes.

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About the authors

Elena A. Gantsova

Petrovsky National Research Centre of Surgery; Peoples’ Friendship University of Russia

Author for correspondence.
Email: gantsova@mail.ru
ORCID iD: 0000-0003-4925-8005
SPIN-code: 6486-5795
Russian Federation, Moscow; Moscow

Oxana V. Serova

Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Email: oxana.serova@gmail.com
ORCID iD: 0000-0002-4941-8913
SPIN-code: 8459-4496

Cand. Sci. (Chemistry)

Russian Federation, Moscow

Igor E. Deev

Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Email: deyevie@gmail.com
ORCID iD: 0000-0002-2041-808X
SPIN-code: 3242-5528

Dr. Sci. (Biology)

Russian Federation, Moscow

Andrey V. Elchaninov

Petrovsky National Research Centre of Surgery; Peoples’ Friendship University of Russia; Kulakov Research Center for Obstetrics, Gynecology and Perinatology

Email: elchandrey@yandex.ru
ORCID iD: 0000-0002-2392-4439
SPIN-code: 5160-9029

Dr. Sci. (Medicine), Associate Professor

Russian Federation, Moscow; Moscow; Moscow

Timur H. Fatkhudinov

Petrovsky National Research Centre of Surgery; Peoples’ Friendship University of Russia; Kulakov Research Center for Obstetrics, Gynecology and Perinatology

Email: fatkhudinov_tkh@pfur.ru
ORCID iD: 0000-0002-6498-5764
SPIN-code: 7919-8430

Dr. Sci. (Medicine), Professor

Russian Federation, Moscow; Moscow; Moscow

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2. Fig. 1. Kidney cryosection morphology in mice: a — wild-type mouse under normal conditions; b — wild-type mouse under bicarbonate load; c — INSRR knockout mouse under normal conditions; d — INSRR knockout mouse under bicarbonate load. Hematoxylin and eosin staining, magnification ×20; scale bar — 100 μm.

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3. Fig. 2. Morphometric characteristics of kidneys: a — relative parenchymal thickness (μm); b — relative glomerular area (μm2); c — collecting duct lumen diameter (μm). Group labels: WT_H2O — wild-type mice under normal conditions; WT_NaHCO3 — wild-type mice under alkaline load; KO_H₂O — INSRR knockout mice under normal conditions; KO_NaHCO3 — INSRR knockout mice under alkaline load. Data are presented as mean ± standard deviation; * — statistically significant differences, p < 0.05.

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4. Fig. 3. Immunohistochemical analysis of CD86-positive macrophages in mouse kidneys: a — wild-type mouse under normal conditions; b — wild-type mouse under bicarbonate load; c — INSRR knockout mouse under normal conditions; d — INSRR knockout mouse under bicarbonate load. Green fluorescence — CD86-positive macrophages (fluorescent dye FITC); blue fluorescence — cell nuclei stained with DAPI. Magnification ×20; scale bar — 50 μm.

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5. Fig. 4. Immunohistochemical analysis of CD206-positive macrophages in mouse kidneys: a — wild-type mouse under normal conditions; b — wild-type mouse under bicarbonate load; c — INSRR knockout mouse under normal conditions; d — INSRR knockout mouse under bicarbonate load. Green fluorescence — CD206-positive macrophages (fluorescent dye FITC); blue fluorescence — cell nuclei stained with DAPI. Magnification ×20; scale bar — 50 μm.

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6. Fig. 5. Number of CD206-positive macrophages in mouse kidneys. Group labels: WT_H2O — wild-type mice under normal conditions; WT_NaHCO3 — wild-type mice under alkaline load; KO_H2O — INSRR knockout mice under normal conditions; KO_NaHCO3 — INSRR knockout mice under alkaline load. Data are presented as mean ± standard deviation.

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