Download Mark Chalkley - ACVP/STP Coalition

Neuronal CeroidLipofuscinosis in Two Cats
MARK CHALKLEY – 3rd YEAR ANATOMIC
PATHOLOGY RESIDENT
History & Signalment
• Similar history and signalment for both case #1 and
#2:
- Both between 1 or 2-years old male DSH cats
- Submitted to animal shelters as strays with no
previous history
- Prolonged, progressive history of visual
dysfunction (running into walls, difficulty tracking
moving objects) and neurological deficits (falling,
frantic running, behavior abnormalities)
• Both euthanized and submitted for necropsy
Case #1
Control
Cerebral Cortex
Normal
Case #2
H&E
H&E
H&E
LFB/
H&E
AF
PAS
Immunofluorescence
• Normal and abnormal
H&E
GFAP
Normal
Case #2
Lesion Distribution
Anatomic Location
Lipopigment/Ceroid
Neuronal/ Cell Loss
Gliosis
1
2
1
2
1
2
Cerebral cortex
+++
++
+++
++
+++
++
Thalamus
+
++
+
+
+
+
Hypothalamus
+
+
+
+
+
+
Basal Nuclei
+
++
+
+
+
+
Hippocampus
+++
+++
+
+
++
+
Cerebellum
++
+
++
+
++
+
Spinal Cord
+
++
+
+
+
+
Retina
+
+
+++
++
-
-
Extra-CNS tissues
-
-
-
-
-
-
* + = mild, ++ = moderate, +++ = severe; 1 = Case No. 1; 2 = Case No. 2
Disease
Cause
Organs affected
1. CeroidLipofuscinoses
Unknown or
lysosomal
protein defects
Predominantly
cerebrum,
cerebellum & retina
in cats but can be in
many tissues
2. Gangliosidoses
Lysosomal
enzyme
deficiency
CNS, PNS, liver,
kidney, lymph n.,
cardiac Purkinje
cells
3. Glucocerebrosidoses
4. Mannosidosis
(glycoproteinosis)
5. Mucopolysaccharidoses
(MPS VII in cats)
Glucocerebrosidase
deficiency
Lysosomal α or β
mannosidase or
α-L-fucosidase
deficiency
Various enzyme
deficiencies
CNS, macrophages
(esp liver, LNs)
Many cells (neurons,
epithelium,
endothelium,
mesenchyme)
Predominantly bone
and other connective
tissues, neurons
Storage
material
Special
stains
Subunit c of
mitochondrial
ATPase,
amyloid-β,
precursor
protein, SAPs
Magneta
with PAS
LFB+
Sudan
black+
Autofluorescent
Glycosphingolipids
PAS+
LFB+
Sudan
black+
Glucocerebrosides
PAS+ in
macs
PAS – in
neurons
LFB+
Carbohydrate
(saccharides
with mannose
residues)
PAS+,
diastase
resistant,
Variably
alcian
blue+
Glucoaminoglycan
Variable
PAS+
LFB+
Sudan
black+
Autofluorescent
Lectin
histochemistry
EM
Variable to
nil reactivity
Curvilinear
bodies
Fingerprint
profiles
Granular
osmophilic
deposits
O-linked
Membranous
cytoplasmic
bodies and/or
zebra bodies
O-linked
Twisted,
branching
tubules or
zebra bodies in
neurons
N-linked
Membrane
bound
vesicles, with
floccular
material & fine
membrane
stacks
N-linked
Empty
vesicles, or
floccular
material /zebra
bodies
Ceroid-lipofuscinosis
Gangliosidoses
Sphingolipidoses &
Mucopolysaccharidoses
Mannosidoses
Diagnosis
Brain, spinal cord, eyes:
Degenerative lysosomal storage encephalomyelopathy and retinopathy, widespread, marked,
chronic, with neuronal loss and astrogliosis
- Consistent with neuronal ceroid-lipofuscinosis
(NCL)
Lysosomal Storage Diseases
(LSD)
What is LSD anyway?
• LySergic acid Diethylamide
• Lumpy Skin Disease
• Lichenoid-pSoriasiform
Dermatoses
• Louisiana School for the
Deaf
NCL in Animals and Humans
•
•
•
•
•
•
Four main variants – infantile, late-infantile, juvenile, adult
160 mutations in 10 human genes (CLN1-10)
Lead to defects in 8 known proteins
Gene defects also recognized in dogs, sheep and cattle
Similar distribution and character of lesions & material
Main storage material in animals and humans - subunit c
of mitochondrial ATP synthase (SCMAS) or
sphingolipid activator proteins A and D (SAPs)
• A correlation between the age of clinical onset of disease,
ultrastructure of the storage material and the variants of
NCL has not been established in animals
Characteristic
Lipofuscin
Ceroid
Lipofuscin vs Ceroid
Circumstance of deposition
Aging
Pathological
Location
Lysosome
Lysosome
Positive
Positive
Protein
Heterogenous mix, amyloid-β protein
Subunit c of mitochondrial ATPase, amyloid-β,
precursor protein, saposins
Lipids
Triglycerides, FFA, dolichols
Phosphorylated dolichols, phospholipids, neutral
lipids
Carbohydrates
Dolichol-linked oligosaccharides
Dolichol-linked oligosaccharides
Metals
Fe, Cu, Al, Zn, Ca, Mn
Predominantly Fe
Sudan Black
Positive
Positive
Luxol fast blue
Positive
Positive
PAS
Positive
Positive
Concanavalin A
Positive
Positive
Agglutinin
Negative
Positive
Granular osmophilic deposits
Positive
Positive
Fingerprint profiles
Negative
Positive
Curvilinear bodies
Negative
Positive
Autofluorescence
Storage components
Histochemistry (including lectins)
Ultrastructure
*Table adapted from SS Seehafer, DA Pearce, Neurology of Aging 27 (2006), p580.
Mechanisms of Ceroid Oxidation
Accumulation
Autophagy
Seehafer S & Pearce D (2007)
Lysosomal
Function
Acknowledgements
• ACVP/STP coalition fellowship
• Pfizer & Dr Peter Schmidt
• University of Minnesota –
- Dr Anibal Armien & Dr Gerry O’Sullivan
- Electron microscopy laboratory
Questions?