Absorption capacity of renal proximal tubular cells studied by combined injections of YFP and GFP in Rana temporaria L

Np, Prutskova; Seliverstova, Elena V.

doi:10.1016/j.cbpa.2013.05.022

Cited by 14 publications

(9 citation statements)

References 33 publications

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“…In our previous study, the involving cubilin and megalin in the uptake and vesicular transport of yellow fluorescent protein (YFP) in the proximal tubule cells of the R. temporaria kidney was shown. 12 As regards other lower vertebrates, similar data on megalin- and cubilin-dependent endocytosis were obtained only for the zebrafish pronephros. 21 The endocytic uptake and intracellular accumulation in the mammalian proximal tubules were first described for larger proteins (albumin, hemoglobin, horseradish peroxidase), but later it was shown that small proteins such as growth hormone, lysozyme, and insulin are absorbed by the same way.…”

Section: Introductionmentioning

confidence: 55%

Receptor-mediated endocytosis of lysozyme in renal proximal tubules of the frog Rana temporaria

Seliverstova

2015

Eur J Histochem

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The mechanism of protein reabsorption in the kidney of lower vertebrates remains insufficiently investigated in spite of raising interest to the amphibian and fish kidneys as a useful model for physiological and pathophysiological examinations. In the present study, we examined the renal tubular uptake and the internalization rote of lysozyme after its intravenous injection in the wintering frog Rana temporaria using immunohisto- and immunocytochemistry and specific markers for some endocytic compartments. The distinct expression of megalin and cubilin in the proximal tubule cells of lysozyme-injected frogs was revealed whereas kidney tissue of control animals showed no positive immunoreactivity. Lysozyme was detected in the apical endocytic compartment of the tubular cells and colocalized with clathrin 10 min after injection. After 20 min, lysozyme was located in the subapical compartment negative to clathrin (endo-somes), and intracellular trafficking of lysozyme was coincided with the distribution of megalin and cubilin. However, internalized protein was retained in the endosomes and did not reach lysosomes within 30 min after treatment that may indicate the inhibition of intra-cellular trafficking in hibernating frogs. For the first time, we provided the evidence that lysozyme is filtered through the glomeruli and absorbed by receptor-mediated clathrin-dependent endocytosis in the frog proximal tubule cells. Thus, the protein uptake in the amphibian mesonephros is mediated by megalin and cubilin that confirms a critical role of endocytic receptors in the renal reabsorption of proteins in amphibians as in mammals.

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Section: Introductionmentioning

confidence: 55%

Receptor-mediated endocytosis of lysozyme in renal proximal tubules of the frog Rana temporaria

Seliverstova

2015

Eur J Histochem

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“…When two proteins were injected in sequence, one before the other or vice versa, the second protein can be colocalized with the first protein but also located in individual endosomes ( Figure 4K) because most of the vesicles containing the first protein moved from the apical cytoplasm to other cell areas. Effect of combined injections did not depend on the order of GFP and YFP introduction [18]. Therefore, the total result is shown below (Figure 6A).…”

Section: Effects Of Combined Protein Injections and Previous Protein mentioning

confidence: 84%

“…In our studies, the expression of endocytic receptors in PT cells of the frog mesonephros was revealed after injections of YFP [18] and lysozyme [17] using polyclonal antibodies against megalin and cubilin. In 15-30 min, absorbed YFP was colocalized with immunolabeled megalin or cubilin in apical endocytic vesicles (Figure 4A-D).…”

Section: Molecular and Cellular Mechanisms Of Endocytosis In The Amphmentioning

confidence: 85%

“…The specific fluorescence intensity, maximum fluorescence, and fluorescence density increased in response to increasing doses of GFP and a high positive correlation was revealed. Reabsorption of fluorescent proteins was also time dependent [15,18]. With increasing time after injection, there was an accumulation of vesicles with GFP or YFP and a movement of some fluorescent endocytic vesicles from the apical cytoplasm to perinuclear and basal areas.…”

Section: Protein Uptake Pattern In the Amphibian And Rat Kidney And Qmentioning

confidence: 94%

“…As YFP and GFP are filtered and absorbed in the kidney in the same way, these proteins may be competitive in the absorptive process. We examined the uptake and intracellular traffic of both GFP and YFP under different conditions for competitive absorption in vivo after simultaneous and sequential introduction of equal amounts of these proteins [18,19]. After simultaneous introduction of GFP and YFP, predominantly colocalized fluorescent signals indicated accumulation of both proteins in the same endocytic vesicles.…”

Section: Effects Of Combined Protein Injections and Previous Protein mentioning

confidence: 99%

See 2 more Smart Citations

Protein Reabsorption in the Amphibian Kidney: Comparative and Evolutionary Aspects

Seliverstova¹,

Np²

2018

Evolutionary Physiology and Biochemistry - Advances and Perspectives

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Protein reabsorption in the renal proximal tubule (PT) is a vitally important process which prevents the loss of filtered proteins and provides their participation in subsequent metabolism. Despite considerable changes in renal structure and function in the process of evolution, very little is known about the functional similarities or specifics of tubular protein reabsorption in the kidney of lower vertebrates compared with the mammalian and human kidney. This article presents an overview of our recent studies on protein reabsorption in the kidney of amphibians, which are used as one of the main animal models for current biological and biomedical research. In frogs, newts, and rats, the absorption capacity of epithelial PT cells was studied after the introduction of green fluorescent protein (GFP), yellow fluorescent protein (YFP), and lysozyme. Molecular mechanisms of receptor-mediated protein endocytosis were also investigated by immunohisto-and immunocytochemistry, electron, fluorescent, and laser scanning confocal microscopy.

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Osmoregulation and Excretion

Larsen

Deaton

Onken

et al. 2014

Comprehensive Physiology

179

141

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The article discusses advances in osmoregulation and excretion with emphasis on how multicellular animals in different osmotic environments regulate their milieu intérieur. Mechanisms of energy transformations in animal osmoregulation are dealt with in biophysical terms with respect to water and ion exchange across biological membranes and coupling of ion and water fluxes across epithelia. The discussion of functions is based on a comparative approach analyzing mechanisms that have evolved in different taxonomic groups at biochemical, cellular and tissue levels and their integration in maintaining whole body water and ion homeostasis. The focus is on recent studies of adaptations and newly discovered mechanisms of acclimatization during transitions of animals between different osmotic environments. Special attention is paid to hypotheses about the diversity of cellular organization of osmoregulatory and excretory organs such as glomerular kidneys, antennal glands, Malpighian tubules and insect gut, gills, integument and intestine, with accounts on experimental approaches and methods applied in the studies. It is demonstrated how knowledge in these areas of comparative physiology has expanded considerably during the last two decades, bridging seminal classical works with studies based on new approaches at all levels of anatomical and functional organization. A number of as yet partially unanswered questions are emphasized, some of which are about how water and solute exchange mechanisms at lower levels are integrated for regulating whole body extracellular water volume and ion homeostasis of animals in their natural habitats. © 2014 American Physiological Society.

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Absorption capacity of renal proximal tubular cells studied by combined injections of YFP and GFP in Rana temporaria L

Cited by 14 publications

References 33 publications

Receptor-mediated endocytosis of lysozyme in renal proximal tubules of the frog Rana temporaria

Receptor-mediated endocytosis of lysozyme in renal proximal tubules of the frog Rana temporaria

Protein Reabsorption in the Amphibian Kidney: Comparative and Evolutionary Aspects

Osmoregulation and Excretion

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