Journal of Evolutionary Biochemistry and PhysiologyJournal of Evolutionary Biochemistry and PhysiologyЖурнал эволюционной биохимии и физиологии0044-45293034-5529The Russian Academy of Sciences64808210.31857/S0044452924030047YXHQKWEXPERIMENTAL ARTICLESЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИResearch ArticleEffect of hypernatremia on protein reabsorption in renal proximal tubules of the lake frog Pelophylax ridibundusВлияние гипернатриемии на реабсорбцию белков в проксимальных канальцах почки озерной лягушки Pelophylax ridibundusPrutskovaN. P.ПруцковаН. П.
Russian Federation
natprut@yandex.ruSeliverstovaE. V.СеливерстоваЕ. В.
Russian Federation
natprut@yandex.ruSechenov Institute of Evolutionary Physiology and Biochemistry RASИнститут эволюционной физиологии и биохимии им. И.М. Сеченова РАН1505202460325426328012025Copyright ©; 2024, Russian Academy of SciencesCopyright ©; 2024, Российская академия наук2024Russian Academy of SciencesРоссийская академия наукhttps://transsyst.ru/0044-4529/article/view/648082

Protein reabsorption in the kidney proximal tubules occurs simultaneously with the transport of ions and water, but little is known about the dependence of receptor-mediated protein endocytosis on water-salt balance changes. The aim of the study was to investigate tubular reabsorption and intracellular vesicular transport of various proteins in a model of hypernatremia in lake frogs (Pelophylax ridibundus). Frogs were injected with hypertonic sodium chloride solution (0.75 M NaCl) 1 hour before injection of green or yellow fluorescent proteins (GFP or YFP), as well as lysozyme. The method of fluorescent immunohistochemistry was used for detection of lysozyme and endocytic receptor megalin in kidney sections. Specimens were investigated using laser scanning confocal microscopy. The intensity of fluorescent signals of proteins and megalin in proximal tubular cells was determined on the images obtained. To study the dynamics of endocytosis, an automated method for quantifying colocalized protein and megalin signals was used. A statistically significant decrease in the reabsorption of GFP, YFP and lysozyme in the proximal tubules after 0.75 M of NaCl injection was found. The accumulation of proteins in the early endocytic compartment and decrease in their entry into late endosomes and lysosomes are shown, that is considered as evidence of a delay in intracellular vesicular transport in hypernatremia. The data obtained were analyzed in connection with changes in blood parameters and kidney activity during osmoregulation, and also with the role of chloride channels in receptor-mediated protein endocytosis. It can be assumed that increased ion transport in the proximal tubules cells in hypernatremia leads to decreased reabsorption capacity of epitheliocytes and delayed intracellular transport of proteins.

Реабсорбция белков в проксимальных канальцах почек происходит одновременно с транспортом ионов и воды, однако мало что известно о зависимости рецептор-опосредованного эндоцитоза белка от изменений водно-солевого баланса. Целью исследования явилось изучение канальцевой реабсорбции и внутриклеточного везикулярного транспорта различных белков в модели гипернатриемии на озерных лягушках (Pelophylax ridibundus). В опытах лягушкам инъецировали гипертонический раствор хлорида натрия (0.75 М NaCl) за 1 ч до инъекции зеленого или желтого флуоресцентных белков (GFP или YFP), а также лизоцима. Для выявления лизоцима и рецептора эндоцитоза мегалина не срезах почек использовали метод флуоресцентной иммуногистохимии. Препараты изучали в лазерном сканирующем конфокальном микроскопе. На полученных изображениях определяли интенсивность флуоресцентных сигналов белков и мегалина в эпителиальных клетках проксимальных канальцев. Для изучения динамики эндоцитоза применяли автоматизированный метод количественной оценки колокализованных сигналов белка и мегалина. Установлено статистически достоверное снижение реабсорбции GFP, YFP и лизоцима в проксимальных канальцах после инъекций раствора NaCl по сравнению с контролем. Показано накопление белков в раннем эндоцитозном компартменте и снижение их поступления в поздние эндосомы и лизосомы, что рассматривается как свидетельство замедления внутриклеточного везикулярного транспорта при гипернатриемии. Полученные данные проанализированы в связи с изменениями показателей крови и деятельности почек в процессе осморегуляции, а также с ролью хлоридных каналов в рецептор-опосредованном эндоцитозе белков. Можно полагать, что усиление эпителиального транспорта ионов в проксимальных канальцах в условиях гипернатриемии приводит к снижению способности эпителиоцитов к реабсорбции белков и ингибированию их внутриклеточного транспорта в процессе эндоцитоза.

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