Gentamicin incorporation along the nephron: Autoradiographic study on isolated tubules

Vandewalle, Alain; Farman, Nicolette; Morin, Jean–Paul; Fillastre, J P; Hatt, P. Y.; Bonvalet, Jean-Paul; Gastineau, with the technical assistance of M.; Wanstok, F

doi:10.1038/ki.1981.50

Cited by 75 publications

(27 citation statements)

References 15 publications

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“…Our results support the evidence that gentamicin is incorporated into proximal tubular cells via the brush border membrane, as previously shown by autoradiographic studies (3,15,17). In fact, 10 min after gentamicin injection, gold particles were found on the brush border membrane and on the membranes of endocytic apical vesicles.…”

Section: Discussionsupporting

confidence: 80%

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Subcellular distribution of gentamicin in proximal tubular cells, determined by immunogold labeling

Beauchamp

Gourde

Bergeron

1991

Antimicrob Agents Chemother

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The subcellular distribution of gentamicin in rat renal proximal tubular cells was evaluated by immunogold labeling. The distribution of the drug was monitored from 10 min to 10 days following single (40 mg/kg of body weight) and multiple (5 and 20 mg/kg/12 h) injections of gentamicin. Animals were killed on day 11, and cubes of renal cortex tissue were fixed overnight in cold phosphate-buffered glutaraldehyde (0.5%), dehydrated in ethanol, and embedded in Araldite 502 epoxy resin. Ultrathin sections were made and incubated with sheep antigentamicin and then with protein A-gold (15 nm) complex. At 10 min after a single injection, the labeling was found over the brush border membrane and over the membranes of endocytic apical vesicles of proximal tubular cells. After 1 h, a similar distribution was observed and the labeling was also seen over small lysosomes located close to the brush border membrane. At 24 h, gold particles were found over large lysosomes of proximal tubular cells. Following 10 days of treatment, lysosomes of proximal tubular cells were densely labeled with gold particles. The labeling was distributed uniformly over the lysosomes, although a lower density of labeling was observed over the myeloid bodies inside the lysosomes. Necrotic proximal tubular cells showed labeling over intact lysosomes and also in the cytoplasms of the cells, in the mitochondria, and in the nucleoli. The various control experiments demonstrated the high specificity of these results. The present immunocytochemical study better documents the subcellular disposition of gentamicim in proximal tubular cells, as previously evaluated by subcellular fractionation and autoradiography. This technique will be useful for better understanding the relationship between drug disposition and drug-induced toxicity.The nephrotoxicity of aminoglycosides has been extensively investigated, but the mechanism by which these drugs induce cellular disturbances and necrosis remains to be elucidated. These drugs, which are essentially eliminated by glomerular filtration, are partially reabsorbed by proximal tubular cells (15). They concentrate into lysosomes (7), where they induce a lysosomal phospholipidosis (9) characterized by the appearance of myeloid bodies (8).The incorporation of aminoglycosides within the lysosomes of renal cortical cells has been demonstrated by subcellular fractionation (16). The binding of aminoglycosides to brush border membranes and their subsequent accumulation into lysosomes of proximal tubular cells have been previously shown by autoradiography (3, 15). Moreover, Giurgea-Marion et al. (7), by using subcellular fractionation, have observed that gentamicin remains associated with the lysosomes of proximal tubular cells throughout a 9-day treatment.The objective of the present study was

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Section: Discussionsupporting

confidence: 80%

“…The uptake of aminoglycosides by proximal tubular cells has been demonstrated by different approaches such as micropuncture (11,14) and autoradiographic experiments (3,15,17). The association of aminoglycosides with lysosomes of cortical renal cells has been shown by subcellular fractionation (16).…”

Section: Discussionmentioning

confidence: 99%

Subcellular distribution of gentamicin in proximal tubular cells, determined by immunogold labeling

Beauchamp

Gourde

Bergeron

1991

Antimicrob Agents Chemother

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“…This renal cortical accumulation reflected the uptake of aminoglycosides exclusively inside proximal tubular cells. Gentamicin has been demonstrated by various techniques to undergo reabsorption from the lumens of the proximal tubules via the brush border (2,8,13,26). There is no evidence to suggest that in the intact in vivo kidney, aminoglycosides enter the cells directly via the basolateral or antiluminal membranes (2,18,28).…”

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confidence: 99%

Gentamicin, netilmicin, dibekacin, and amikacin nephrotoxicity and its relationship to tubular reabsorption in rabbits

Brion¹,

Barge²,

Godefroy³

et al. 1984

Antimicrob Agents Chemother

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The role of the tubular reabsorption of aminoglycosides in nephrotoxicity was considered. The tubular reabsorption rate, fractional reabsorption, and net balance, expressed as the excreted to infused aminoglycoside ratio, were concomitantly studied in male rabbits by continuous infusion of gentamicin, netilmicin, dibekacin, and amikacin. Aminoglycoside nephrotoxicity was evaluated by creatinine levels in serum and pathological renal damage after 14 days of a low-or high-dose regimen, comprising either eight, hourly intramuscular injections of gentamicin, netilmicin, or dibekacin (4 mg/kg) or amikacin (16 mg/kg); twelve, hourly intramuscular injections of gentamicin, netilmicin, or dibekacin (15 mg/kg) or amikacin (60 mg/kg); or injections of saline for the control group. Aminoglycosides exhibited three degrees of tubular reabsorption: gentamicin had the highest, netilmicin had the lowest, and dibekacin and amikacin had intermediate degrees of reabsorption. Nephrotoxicity associated with alteration in renal histology was observed with gentamicin and, to a lesser extent, with dibekacin in the high-dose regimen. No nephrotoxicity was noted with netilmicin or amikacin compared with the control group. Concentrations of the aminoglycosides in renal cortex and serum were not predictive of renal toxicity. Except for amikacin, which appeared to exhibit the lowest intrinsic renal toxicity, nephrotoxicity was correlated with the tubular reabsorption of each aminoglycoside. It was concluded that aminoglycoside renal toxicity can be determined by two major factors: importance of transport into tubular cells and intrinsic intracellular toxicity.Aminoglycoside nephrotoxicity has been the subject of many clinical, pharmacological, and biochemical investigations (16,20,28). Although the nephrotoxic potential of these drugs varies from one species to another, similar pathological patterns have generally been observed in both humans and animals. Morphological lesions, mainly of the proximal tubular cells (10,25), have been demonstrated by light and electron microscopy. In animal models, differences in aminoglycoside nephrotoxicity were identified some time ago, particularly when, on a weight basis, the doses were 10 to 25 times those used in humans. Under these conditions, gentamicin exhibited greater nephrotoxicity than netilmicin, tobramycin, dibekacin, or amikacin (11,14,25,27). The available evidence indicates that the nephrotoxic potential of aminoglycosides cannot be correlated with peak or trough levels in serum (3). Aminoglycoside accumulation in rat parenchyma was recently correlated with gentamicin and tobramycin nephrotoxicity (1). However, the importance of tissue accumulation per se in the pathogenesis of nephrotoxicity remains uncertain (14,25,27). By using a mathematically derived pharmacokinetic model, Schentag et al. (22) concluded that the extent of timed renal cortical accumulation correlated with clinical toxicity in humans. This renal cortical accumulation reflected the uptake of aminoglycosides exclusively in...

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“…Re-uptake can occur via megalin-mediated endocytosis in the proximal tubule [6]. This leads to increased concentration of AGs in proximal tubule cells and, therefore, the particular susceptibility of the kidney to AG toxicity [7]. Thus, mitochondrial dysfunction in the proximal tubule is the expected consequence of AG toxicity, and this is, indeed, concordant with the clinical findings: AG-induced kidney failure is typically non-oliguric, with evidence of proximal tubular dysfunction and damage (reviewed in [8]) and occasionally frank renal Fanconi syndrome [9].…”

Section: Aminoglycosides and The Kidneymentioning

confidence: 99%

Cystic fibrosis, aminoglycoside treatment and acute renal failure: the not so gentle micin

2009

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Aminoglycosides have a wide spectrum of gramnegative anti-bacterial activities and are available at low cost, which makes them commonly used drugs, especially for patients with cystic fibrosis (CF), who often suffer from chronic lung infections from Pseudomonas aeruginosa. Unfortunately, this treatment seems to have resulted in an increased incidence of acute renal failure (ARF) in patients with CF. A recent case-control study investigated risk factors for ARF in CF patients and suggested intravenous use of gentamicin as the prime culprit. Moreover, in most cases, at least one other risk factor, such as CF-related diabetes, pre-existing renal failure, dehydration or concurrent use of other nephrotoxic drugs, was present. We comment on the renal handling of aminoglycosides and the possible mechanisms of toxicity, as well as strategies for risk minimisation.

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Gentamicin incorporation along the nephron: Autoradiographic study on isolated tubules

Cited by 75 publications

References 15 publications

Subcellular distribution of gentamicin in proximal tubular cells, determined by immunogold labeling

Subcellular distribution of gentamicin in proximal tubular cells, determined by immunogold labeling

Gentamicin, netilmicin, dibekacin, and amikacin nephrotoxicity and its relationship to tubular reabsorption in rabbits

Cystic fibrosis, aminoglycoside treatment and acute renal failure: the not so gentle micin

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