Protein endocytosis by a kidney tubule suspension: metabolic requirements.
Endocytosis in the renal tubular cell is a permanent process serving the role of saving nitrogen from plasma peptides that are continuously cleared away by kidney glomerulus. Since small proteins appear in urine after strenuous exercise, it was hypothesized that renal ischemia impairs the tubular endocytic reabsorption of proteins. The aim of this paper is to describe a simple in vitro model of renal endocytosis and to use it in studies of endocytic metabolic requirements. The results show that rabbit renal proximal tubules in suspension are able to take up 125I-lysozyme, as well as RITC-lactalbumin. The fluorescent protein was taken up only by the ends of the everted tubule fragments, and accumulated into intracellular vesicles, demonstrating the luminal pathway of endocytosis. The amount of 125I-lysozyme taken up was equivalent to that taken up by isolated perfused tubules (Nielsen et al. (1986) Am. J. Physiol. 251, F822-F830). Anoxia decreased 12-fold the intracellular accumulation of 125I-lysozyme; however, the time-course of inhibition shows that only the late steps of endocytic accumulation are energy-dependent. Substrate deprivation studies suggest a specific role of glucose to sustain endocytosis. Lastly, renal uptake of 125I-lysozyme was shown to be strongly depressed by chloroquine, an alkalinizing agent of endosomes and lysosomes. We conclude that (1) renal tubules in suspension are a satisfactory model for endocytic studies in kidney; (2) suppressing oxygen and substrate supplies to kidney impairs endocytic tubular reabsorption of proteins.
Simonnet
H
H
Laboratoire de Physiopathologie Rénale et Métabolique, CNRS EP 18, Faculté de Médecine Alexis Carrel, Lyon, France.
Gauthier
C
C
Gire
V
V
Baverel
G
G
D000409:Alanine / Q000378:metabolism; D000818:Animals; D015687:Cell Hypoxia; D002738:Chloroquine; D004705:Endocytosis; D005947:Glucose / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D007457:Iodine Radioisotopes; D007687:Kidney Tubules, Proximal / Q000378:metabolism*; D007768:Lactalbumin / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D011817:Rabbits; D012235:Rhodamines
D007457:Iodine Radioisotopes; D011506:Proteins; D012235:Rhodamines; D002738:Chloroquine; D007768:Lactalbumin; C027755:rhodamine isothiocyanate; D009113:Muramidase; D005978:Glutathione; D005947:Glucose; D000409:Alanine