Structure and Function of the

Kidney
The functional unit of the kidney is the nephron
The major functions of the kidney are to maintain
extracellular fluids, to eliminate wastes resulting
from normal metabolism, and to excrete
xenobiotics and their metabolites
Mammalian kidneys have 10,000-1,000,000
nephrons per kidney

Renal toxicology

Structure and Function of the

Kidney (cont)
The glomerulus yields an ultrafiltrate of plasma that
represents 20% of the renal blood flow, ie. 2-3% of cardiac
output
Endothelial surface is negatively charged and contains
fenestrae
The glomerular basement membrane is sandwiched
between the epithelial cells and contains anionic
sialoglycoproteins, glycoproteins and collagen IV
The mesangium provides support
The outer capsule is Bowmans capsule

Structure and Function of the

Kidney (cont)
The tubule resorbs greater than 99% of the glomerular
filtrate
The proximal tubule has extensive resorption and selective
secretion (convoluted - S1 and S2, straight - S3). S2 is
primary site for low MW protein resorption and S3 is
primary site for P450.
Thin loop of Henle - resorption of fluids
Distal tubule - resorption of fluids and acid-base balance
Collecting duct - resorption of fluids, antidiuretic hormone
and acid-base balance

Medullary
ray

P2
P2
Cortical
labyrinth
P1 / P2

P1
P3

Cortex

Outer
Medulla

P3
DT

Outer
Stripe
Inner
Stripe

TLH

CD

Inner
Medulla

Short et al., Laboratory Investigation,564-577 (1987).

Structure and Function of the

Kidney (cont)
Produces erythropoietin, which regulates RBC
production
Hydroxylates 25-OH-cholecalciferol (vitamin D
metabolite), to promote bone resorption and
calcium and phosphorus absorption from the gut
Releases renin to regulate the peripheral reninangiotensin-aldosterone system (juctaglomerular
apparatus)

Assessment of Kidney Function:

Morphologic Evaluation

Urinalysis
Gross evaluation of the kidney at necropsy
Histopathology of the kidney
Electron microscopy of the kidney

Assessment of Kidney Function:

Urinalysis

Proteinuria - indicates glomerular damage

Glycosuria - indicates tubular damage
Urine volume and osmolarity
pH
Enzymes - indicates tubular damage
Microscopic examination - casts, crystals,
bacteria, etc.

Assessment of Kidney Function:

Blood Chemistries

Blood urea nitrogen (BUN)

Creatinine
Electrolytes - Ca, Mg, K, P
Glomerular filtration rate - determines the
clearance of inulin, creatinine and BUN
Renal clearance - measures the clearance of
p-aminohippuric acid by filtration and
secretion

Glomerular Disease: Toxicities due

to Alteration of Anionic Charge
Hexadimethrine - polycationic molecule
reduces anionic charge, which permits
escape of anionic molecules such as
albumin and IgG
Polynucleoside of puromycin - damages
epithelial foot processes

Glomerular Disease:
Immune Complex Disease
Anti-GBM mediated glomerulonephritis is
induced by heterologous antibodies
Antibodies due to exogenous antigens cationized molecules such as lysozyme, IgG
and BSA bind to anionized surfaces;
Concanavalin A binds to sugars in the GBM

Glomerular Disease:
Immune Complex Disease (cont)
Deposition of circulating immune complexes
Drug or toxin-induced T-cell dependent polyclonal
B-cell activation - mercury in Brown Norway rats
Unknown mechanism - gold salts, Dpenicillamine, hydralazine
Antibodies to heterologous proteins - safety
evaluations of recombinant proteins in laboratory
animals

Nephrosis:
Damage to the renal tubule
Halogenated hydrocarbons - chloroform,
hexachlorobutadiene, trichloroethylene,
dibromochloropropane, & bromobenzene
Heavy metals - cadmium, mercury & lead
Antibiotics - cephalosporins &
aminoglycosides
Mycotoxins - ochratoxin A & citrinin
Ethylene glycol
Antineoplastic drugs - cisplatinum
Alpha2u-globulin nephropathy

Haloalkane Nephrosis
Chloroform is metabolized by P450 to an
electrophile, phosgene, which is a potent
cytotoxicant.
Carbon tetrachloride is metabolized to free radicals
and phosgene.
P450 is localized in the proximal tubule.
This results in nephrosis with necrosis, enzyme,
glucose and protein excretion in urine, and increased
BUN and creatinine concentrations in serum.

Haloalkene Nephrotoxicity
1,1-Dichloroethylene, trichloroethylene and
tetrachloroethylene are metabolized by
P450 to electrophilic metabolites and or free
radicals.
These metabolites can be cytotoxic and/or
genotoxic.
Nephrotoxicity is exacerbated when
glutathione is depleted.

Glutathione-mediated Nephrosis
Glutathione conjugates of haloalkanes can
form episulfonium ions.
Primary route for 1,2-dichloroethane, 1,2dibromoethane and 1,2-dibromo-3-chloropropane.
These can alkylate macromolecules and
cause cytotoxicity and genotoxicity.

Cystine Conjugate -lyase Activation

Stable cystine conjugates from glutathione
can be formed in the liver from trichloroethylene, tetrafluoroethylene and hexachlorobutadiene and transported to the
kidney.
They are further metabolized by -lyase in
the kidney to generate reactive thiols.

Biotransformation of Trichloroethylene
H
Cl

Cytochrome
P450
Fe

C C

Cl
Cl

+ GSH
GSH-transferase

H
Cl

C C

H O Cl
C C
Cl
Cl

Cl

SG

Cl
-Lyase

H
Cl

C C S

Chlorothioketene

C C

Cl
S CH2

NH3+
CH
COO-

Trichloroethylene-associated
renal cell carcinomas
Highly exposed workers exhibit nephrosis and an increased
incidence of renal cancer
TCE and other renal neoplasms have a high incidence of
mutations in the von Hipple-Lindau (VHL) tumor
suppressor gene. This requires mutation of one allele and
deletion of the other
TCE mutations include a hot spot at nucleotide 454 (CT)
VHL gene is located at 3p25
3p25.5 is a fragile site at the chromosome telomere
ogg1 mutations are also associated with renal cancer and it
is also located at 3p25

Lead Nephropathy
Lead induces acute nephrosis of the proximal tubule
and affects blood pressure to the kidney.
Low molecular weight proteins bind Pb2+, leading
to resorption by endocytosis.
Cellular and mitochondrial swelling, karyomegaly,
mitosis and intranuclear inculsion bodies are seen.
Decreased tubular resorption of glucose, phosphate
and amino acids +/- proteinuria
Chronic exposure causes tubulo-interstitial disease.

Cadmium Nephrotoxicity
Metalothionein (Mt) is a low molecular weight protein that
transports cadmium (Cd) to the kidney.
Mt is taken up in the lysosomes and Cd is released. Cd t
is 10-30 years in humans.
Free Cd stimulates synthesis of renal Mt, which binds Cd.
When renal Mt is depleted, toxicity occurs in S1 and S2.
Proteinuria, calciuria, aminoaciduria and glucosuria are
seen.
Chronic exposure leads to tubulo-interstitial disease. There
appears to be a threshold of 200 ppm Cd in the kidney
before renal disease occurs.

Mercury Nephropathy
Hg2+ and methylmercury cause damage to the S3 and
S2/S3 segments of the proximal tubule.
Hg2+ causes damage to the cell membrane and
mitochondria, and loss of cellular control over
intracellular Ca++.
Chronic exposure results in anti-GBM and immunecomplex disease of the kidney.
Methylmercury is highly lipid soluble and
concentrates in the proximal tubule, causing damage
to mitochondria and lysosomes.

Chromium Nephropathy
Cr+6 causes acute nephrosis of S1 and S2.
It damages the brush border, causing
decreased absorption of proteins, glucose
and amino acids.
Large membranous myeloid bodies form as
a result of disrupted lysosomal function.
Decreased glomerular filtration leads to
increased BUN and creatinine.

Ethylene Glycol Nephropathy

Toxicity first discovered in humans when
ethylene glycol was used as a drug solvent.
Causes acute renal toxicity in animals after
exposure to anti-freeze.
Ethylene glycol is metabolized to oxalic
acid.
Calcium oxylate crystals form in the lumen
of the tubules.

Ethylene Glycol Toxicity

Renal Papillary Necrosis

Caused by non-steroidal anti-inflammatory
drugs phenacetin and acetaminophen.
Highest concentration of drugs are in the
renal papilla.
Prostaglandin hydroperoxidase is highest in
the medulla and is thought to metabolize
these drugs to reactive quinoneimines.

Urinary Bladder Toxins

Chemicals that cause bladder stones and
calcium phosphate precipitates frequently
cause bladder cancer in rodents.
Aromatic amines, tobacco smoke and
parasitic infections cause bladder cancer in
humans.

Saccharin
Non-genotoxic
Induces bladder epithelial tumors in male rats
at >1% in the diet
Must be administered to neonatal animals
Can act as a promoter for genotoxic agents
Only the sodium salt of saccharin is
carcinogenic
Mechanism requires the formation of calcium
phosphate-containing precipitates