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

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.

Keywords

IRR receptor tyrosine kinase / alkalosis / kidney / macrophages

References

[1] Korotkova DD, Gantsova EA, Goryashchenko AS, et al. Insulin receptor-related receptor regulates the rate of early development in Xenopus laevis. Int J Mol Sci. 2022;23(16):9250. doi: 10.3390/ijms23169250

[2] Deyev IE, Sohet F, Vassilenko KP, et al. Insulin receptor-related receptor as an extracellular alkali sensor. Cell Metab. 2011;13(6):679–689. doi: 10.1016/j.cmet.2011.03.022

[3] Serova OV, Gantsova EA, Deyev IE, Petrenko AG. The value of pH sensors in maintaining homeostasis of the nervous system. Russian Journal of Bioorganic Chemistry. 2020;46(4):369–384. (In Russ.) doi: 10.31857/S0132342320040260 EDN: PYRUCK

[4] Petrenko AG, Zozulya SA, Deyev IE, Eladari D. Insulin receptor-related receptor as an extracellular pH sensor involved in the regulation of acid–base balance. Biochim Biophys. 2013;1834(10):2170–2175. doi: 10.1016/j.bbapap.2012.11.011

[5] Alva S, Divyashree M, Kamath J. et al. A study on effect of bicarbonate supplementation on the progression of chronic kidney disease. Indian J Nephrol. 2020;30(2):91–97. doi: 10.4103/ijn.IJN_93_19

[6] Dubey AK, Sahoo J, Vairappan B, et al. Correction of metabolic acidosis improves muscle mass and renal function in chronic kidney disease stages 3 and 4: a randomized controlled trial. Nephrol Dial Transplant. 2020;35(1):121–129. doi: 10.1093/ndt/gfy214

[7] Di Iorio BR, Bellasi A, Raphael KL, et al. Treatment of metabolic acidosis with sodium bicarbonate delays progression of chronic kidney disease: the UBI Study. J Nephrol. 2019;32(6):989–1001. doi: 10.1007/s40620-019-00656-5

[8] Goraya N, Munoz-Maldonado Y, Simoni J, Wesson DE. Fruit and vegetable treatment of chronic kidney disease-related metabolic acidosis reduces cardiovascular risk better than sodium bicarbonate. Am J Nephrol. 2019;49(6):438–448. doi: 10.1159/000500042

[9] Mannon EC, O’Connor PM. Alkali supplementation as a therapeutic in chronic kidney disease: What mediates protection? Am J Physiol Renal Physiol. 2020;319(6):F1090–F1104. doi: 10.1152/ajprenal.00343.2020

[10] Gantsova EA, Serova OV, Eladari D, et al. A comparative kidney transcriptome analysis of bicarbonate-loaded insrr-null mice. Curr Issues Mol Biol. 2023;45(12):9709–9722. doi: 10.3390/cimb45120606

[11] Chen S, Saeed AFUH, Liu Q, et al. Macrophages in immunoregulation and therapeutics. Signal Transduct Target Ther. 2023;8(1):207. doi: 10.1038/s41392-023-01452-1

[12] Wu H, Yin Y, Hu X, et al. Effects of environmental pH on macrophage polarization and osteoimmunomodulation. ACS Biomater Sci Eng. 2019;5(10):5548–5557. doi: 10.1021/acsbiomaterials.9b01181

[13] Genini A, Mohebbi N, Daryadel A, et al. Adaptive response of the murine collecting duct to alkali loading. Pflugers Arch. 2020;472(8):1079–1092. doi: 10.1007/s00424-020-02423-z

[14] Tobar A, Ori Y, Benchetrit S, et al. Proximal tubular hypertrophy and enlarged glomerular and proximal tubular urinary space in obese subjects with proteinuria. PLoS One. 2013;8(9):e75547. doi: 10.1371/journal.pone.0075547

[15] Chen H, Birnbaum Y, Ye R, et al. SGLT2 inhibition by Dapagliflozin attenuates diabetic ketoacidosis in mice with type-1 diabetes. Cardiovasc Drugs Ther. 2022;36(6):1091–1108. doi: 10.1007/s10557-021-07243-6

[16] Lu YP, Zhang ZY, Wu HW, et al. SGLT2 inhibitors improve kidney function and morphology by regulating renal metabolic reprogramming in mice with diabetic kidney disease. J Transl Med. 2022;20(1):420. doi: 10.1186/s12967-022-03629-8

[17] Liu C, Wang X. Clinical utility of ultrasonographic evaluation in acute kidney injury. Transl Androl Urol. 2020;9(3):1345–1355. doi: 10.21037/tau-20-831

[18] Gupta P, Chatterjee S, Debnath J, et al. Ultrasonographic predictors in chronic kidney disease: A hospital based case control study. J Clin Ultrasound. 2021;49(7):715–719. doi: 10.1002/jcu.23026

[19] Fenton RA, Knepper MA. Mouse models and the urinary concentrating mechanism in the new millennium. Physiol Rev. 2007;87(4):1083–1112. doi: 10.1152/physrev.00053.2006

[20] Gantsova E, Serova O, Vishnyakova P, et al. Mechanisms and physiological relevance of acid-base exchange in functional units of the kidney. PeerJ. 2024;12:e17316. doi: 10.7717/peerj.17316

[21] Amlal H, Ledoussal C, Sheriff S, et al. Downregulation of renal AQP2 water channel and NKCC2 in mice lacking the apical Na+-H+ exchanger NHE3. J Physiol. 2003;553(Pt 2):511–522. doi: 10.1113/jphysiol.2003.053363

[22] Takahashi N, Chernavvsky DR, Gomez RA, et al. Uncompensated polyuria in a mouse model of Bartter’s syndrome. Proc Natl Acad Sci USA. 2000;97(10):5434–5439. doi: 10.1073/pnas.090091297

[23] Lorenz JN, Baird NR, Judd LM, et al. Impaired renal NaCl absorption in mice lacking the ROMK potassium channel, a model for type II Bartter’s syndrome. J Biol Chem. 2002;277(40):37871–37880. doi: 10.1074/jbc.M205627200

[24] Percie du Sert N, Hurst V, Ahluwalia A, et al. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. BMC Vet Res. 2020;16(1):242. doi: 10.1186/s12917-020-02451-y

Funding

Russian Science FoundationРоссийский научный фондRussian Science Foundation(24-45-00031)

Government of the Russian FederationПравительство Российской ФедерацииGovernment of the Russian Federation(123030700110-4)

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