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OVINE CEROID-LIPOFUSCINOSIS
A thesis presented in partial fulfilment
of the requirements for the degree of
Master of Veterinary Science
at Massey University
Antonie Janrnaat
1979
(70%)
to
my
pare n t s
... "Ie hebt
't good edo"'an "..
.
ABSTRACT
Ov ine ceroid-l ipofu scino s i s i s a rare inh e ri ted neu rological
di se a s e which ha s only been di agnosed in one fami l y of the South Hampsh i re
breed o f sheep .
Th i s breed i s o f re la tive recent origin a nd was e s tab l i shed
from a n initia l cro s s between Sou thdcwn and Hampsh ire Down sheep .
Af fected
sheep show loss of vi s ion and behav iou ra l abnorma l i t i e s starting at 1 1 - 1 2
month s o f age , wi th motor dysfunc tion commenc ing soon a fterwa rds.
The
c l i n i c a l s i gns inc r e a s e in sever ity a s the d i s e a s e progre s s e s and unde r
field con d i t ions a f fected anima l s are not expected to l ive b eyond 2
years .
Ovine ceroid-l ipo fu scino s i s i s cha racte ri zed h i s tologica l ly by the
i ntracytopl a smic accumu la tion of PAS and Sudan b l ack po si tive
autof luorescent l ipop igment ma te r i a l in neuron s and a wide variety of
other c e l l type s .
The proce s s l eading to the accumu l a t ion o f l ipopigme n t
seems o n l y t o dama g e n eurons a nd there i s degenera tion a n d l o s s o f
neurons , e specia l ly in the cerebra l cortex a nd the v i sua l neuroepi the l i um
o f the re t ina i . e . retina l a trophy .
reduc t i o n in s i z e a nd we i gh
Gro s s l y , a f fected bra ins show
on average 66% of those of norma l sheep .
U l t ra s tructura l l y , the typ ica l l ipop i gment inc l u s ion i s a round
or ova l body 0 . 2 - 5 . 0 �m in s i ze , o f va rying e l ectron density,
a wide �ariety of memb ranous profi l e s may b e seen .
in which
Some o f the
(
memb ranous patterns have rece ived spec i a l names such as curvi l i n e a r ,
finge rpr int a n d crysta l l o i d .
Pathological examinat ion of l i ver , skin and recta l biopsy mate r i a l
o f l ambs a t 4
-
5
mon ths o f age shows the pre sence o f ac cumul ated
l ipopigment , and i s a means of early d i agno s i s be fore the onse t o f
c l i n i cal s i gn s .
Thi s obs e r va t i on and the fa c t that l ipopigment h a s be e n
demon s t ra ted in a f fe c t ed lambs a t bi rth , show ovine cero i d - l ipofu s c i no s i s
to be a s soc iated w i th a true inborn e rror o f me taboli sm .
The fami ly tree o f a l l a f fe c ted sheep and the resul t s o f s i re­
daughte r mat ings of a hete rozygou s ram show the d i se a se to be inh e r i t e d a s
a s impl e au to soma l rece s s ive tra i t .
The d e l e t e rious gene for ovi ne
c e roid-l ipofusc ino s i s i s u n l ike ly to b e of economic importance to the
sheep industry a s the Sou t h Hampsh i re breed was developed to suppl y
s i r e s for termina l cros se. s a s soc iated with table l amb produc t ion .
Th e obj e c t s of this study were to d e f i n e ov ine ceroid-l ipofu s c i no s i s
i n c l inic a l , patholog i c a l a n d genetic te rms , and to compa re i t w i th
s im i l a r d i se a s e s in man and dome s t i c a n ima l s .
I t is conc l uded that the
ov ine di sease doe s indeed be l ong to the hete rogeneou s g roup o f d i s e a s e s
o f ma n and dome s t i c anima l s known a s Batten ' s d i s ease o r the neuron a l
o r genera l i sed c ero i d - l ipo fu s c i no se s .
Of th e s e the ovine e n t i ty mo s t
c l o s e l y resembl e s the l a t e i n fant i l e and the j uvenile forms of the human
synd rome , and the can ine disea se .
I t i s proposed that ovine c e ro i d -
l i po fu sc ino s i s woul d make a u s e fu l e xperimenta l mode l for Batten ' s
d i s e a se .
ACKNOWL E DGEMENTS
This the s i s re f l ec t s a substantial part o f the rese arch I d id
wh i l e employed as a Re sea rch O f f ic e r in the Depa rtment o f Ve te rina ry
Patho logy and P ub l i c Hea l th a t rla s s ey Un iver s i ty ove r a per iod of 3 yea rs .
I am g ra te fu l to Prof . B . W . Manktelow , as Head o f Depa r tmen t , and to
Dr R . D . Jo l l y ,
as leader o f th e r e s earch team , f i r s t l y for a l lowing me
to u s e the r e s e arch find i n g s i n th i s the s i s and second l y for the
pert i nent advi c e , pa r t i cu l a r l y Dr Jo l l y ' s , pro ffered in the i r r ol e s
a s supervi sor in i t s prepa ra t ion .
A number o f peop l e gave pro f e s s ional
advic e an d technical a s s i s t a nce du ring th e course o f my s tudy i nto
ovine c e r o i d - l ipofusc inos i s .
Mr
D.M . We s t gave generou s l y o f h i s time w i th the c l i n i c a l
exam i na t i on o f the anima l s a n d supervised the an a e s thet ic , surg ical and
po s t-operat ive procedures i nvolved in the taking o f sk i n , l iver and
r e c ta l biops i e s .
He a l so suppl i e d the norma l retinograph
fi gure 2 . 3 as a contro l .
haemogram e st imation s .
u se d in
Dr R . H . Sutton and h i s s t a f f performed the
Ms . K . C a to took the radiog raph s wh ich we re
inte rpreted b y Dr M. O ' Ca l l agha n .
Prepa r a t ion o f t i s su e s for l ight and f luore scent mic ro scopy was
c a r r i ed out by Mr C. Fletche r , Mr R . P . Han sen , Ms . S . L . Ma l l och a nd
Mr s P . M . S l a c k .
coope rat ion .
I thank them for the i r cheerful and f r i endly
Mr A.S. C ra ig and Mr D . Hopcroft o f the Departmen t o f
S c i e n ti f i c a n d Indu s t r i a l Research gave very helpfu l a dvice on the
preparation of t i s sues for e le c t ronmicro scopy and printed the
e l ec t ronmic rograph s .
M r T. Law mos t obl igingly prin te d the o th e r
photographs used i n th i s the s i s .
I am i n debted to Mrs E . Br i s tol
and Al i s o n for typi n g the f i r s t draft of the manu script , and to
Mr s F . S . Wicherts for typ i ng the f i na l copy .
Fina l l y ,
I wou ld l ike to thank the farm and hosp i t a l techn i c a l
s taff for the ir care o f and i n terest i n the anima l s , and espec i a l l y
Mr
A . T . DeCl eene for hi s cheerful and knowl edgeable a s s i s tance w i th
the handl i n g o f the sheep i n the f i e l d , hosp ital and post-mortem
room .
The research pro j ec t i n which I was i nvolved w a s supporte d by
the U n i t e d S tates Na tion a l In s t i tutes o f Health Gra n t No . RO l
NS 1 1 2 3 8 - 0 4 and 5 from the National I n s t i tute of Neuro logic a l Di sord ers
and S troke .
TABLE OF C O NT E NTS
Page
LIST OF TABLES
LIST OF FIGURES
INTRODUCTION
1
CHAPTE R I :
Review - the neurona l ceroid - l ipofu s c i nose s .
CHAPTER I I :
C l inica l f indings .
43
CHAPTE R I I I :
Pathology .
63
CHAPTER IV :
Genetic s .
104
Ge ne r a l d i sc u s s ion .
109
C HAPTER
V:
REFERENCES
2
115
LIST
OF
TABL E S
Table
Page
l. I
Cl a s s i f icat ion o f the neuronal c e roid -l ipofu sc ino s e s .
2.I
C a s e h i s tories of Sou th H ampshire sheep af fected
with c ero i d - l ipofu sc i no s i s.
3.I
64
Di stribut ion o f l ipop i gment i n ceroid - l ipo fu s c i nos i s
o f sheep over 1 2 mon ths o f age .
4.I
46
L i s t o f South Hamp s h i r e s heep wi th ceroid­
l ipofu s c i no s i s that we r e examined pathologic a l ly .
3. II
9
71
Re su l t s o f s i re -daughter ma t ings of a South
H ampshire ram hetero zygous for c eroid- l ipo fu sc i no s i s .
107
LIST
OF
FIGURES
Figu re
2.1
Page
South Hampshire ram , 2 4 months of age ,
52
a f fected w i th c e roid-l ipo fu s c i nosi s .
2.2
Graham-Knol l techn iqu e to d emon strate perox ida s e
act ivity i n sheep neutroph i l s.
A part i a l ly reacted
a nd a comp l ete l y reacted c e l l a re shown .
2.3
55
Retinog raphs o f the ovi ne eye showing attenuat ion
and stra ighten i ng o f retina l ve s s e l s in an a f fected
2 4 months old ram , when compa red to a control sheep .
3.1
57
Dor s a l v i ew o f bra ins f rom two 1 3 months old ewe
hoggets .
The b ra in o f the a f fected sheep i s reduced
in s i ze and shows thinning o f the c e rebral gyri .
The
b ra i n we ight of the a f fe c ted animal is 5 7 . 2 g , compa red
with 8 6 . 2 g for the norm a l control she ep .
3.2
67
Late ral v iew o f brains f rom two 1 3 months o l d ewe
hoggets .
The ce rebrum o f the a f fe cted an ima l shows
reduct ion in s i ze , th i nn i ng of the ce rebra l gyr i and
a s l i ght dor so-ventra l f lattening , when compared with
the b ra i n o f the normal control sheep .
68
Page
Figure
3. 3
Transverse s e c t ion o f the cerebrum of an a f fected
1 3 month s old ewe hogget and of an age and sex­
matched control she ep .
No te r educ ed size o f c e rebrum ,
ma rked thi n n ing o f cortex, th i nning o f corpus callosum
and sepb� pellucidum and e n l a rgement of late r a l
vent r i c l e s i n the a f f e c t e d sheep .
3.4
69
Cerebral cortex of a 1 5 month s o l d South Hampshi re
ewe hogget wi th ce ro i d - l ipofu scinosis showing
neuron s l aden with l ipop i gme n t granul es .
3.5
73
Tha lamus o f a 2 5 mon ths o l d a f fected ram.
Lipopigment granu l e s s ta i n bl ack and in some
neurons appe a r to almo st compl e t e l y f i l l the
cytopla sm .
3.6
73
Cerebra l co rtex of a 2 5 mon ths o l d a f fe cted ram .
The whi te s t ruc ture s are a u tofluore scent
l ipopigme n t granul e s , s i n g l e o r in cluste r s .
3 .7
75
Cerebel l a r cortex o f a 2 5 months old affected
ram .
Inclu s ion s in a Purk i n j e ce l l are nume rous
and show va riation in s i ze .
75
Page
F i gure
3. 8
Cerebral cort e x o f a 2 5 month s old ram with
c ero i d - l ipofusc inosi s , show ing a neuron surrounded
by an increased number of gl i al c e l l s .
Astrocyte s
predominate and may show unusu a l l y shaped nuc l e i.
3.9
Cerebra l cor t i c e s of two approx imat e l y
old South Hampsh ire ewe hogge t s .
15
77
mon ths
The affected
an imal shows an incre ase in number and size of
a s trocyte s , compare d to the control sheep .
3 . 10
77
The h i s tology of the re t i na o f a n 18 months old
South Hampsh i re ewe w i th ceroi d -lipo fuscinos i s ,
compared w i th that o f a norma l sheep .
The a f fected
an ima l shows severe atrophy o f the l ayer o f rod s
'
a nd cone s and the outer nuc l e a r l ayer .
The re t i n a l
p igment ep i the l ium , i nner nucl ear l ayer and
gangl ion cel l l ayer appear normal .
3 . 11
Lymphocyte s from a 25 months o l d a f fected ram .
Note absence o f l ymphocytic vacuolat ion .
3 . 12
78
84
Re sidual body i n cort i c a l n euron showing
l obul ated shape .
84
Page
Figur e
3 . 13
Storage material appare ntly free in the cytoplasm
o f a men inge al cap i l lary endothelial c ell .
Shor t
arrow po ints t o a five - l ayered membrane , wh ich a t
long a rrow seems to b e formed b y the fu sion o f
85
two tr iparti te membrane s .
3 . 14
Part o f l i popigment body i n ret in al neuron showing
granu l a r matrix of vary ing e l ec t ron density , and
five - l ayered membrane s .
The arrow ind i cate s a
85
mye l i n pattern .
3 . 15
P a r t o f res i dual body i n a ventral horn neuron
showing stacks of alte rna t ing dense and l ight
l ine s , with a period i c i ty of approx ima te ly 5 . 2nm .
3 . 16
Panc rea t i c acinar c e l l w i th curv i l i near inc lus ion
bod i e s .
3.17
The cellula r a rch i tecture appears no rma l .
88
Storage body w i thin a c e reb r a l c o r t i c a l neuron
showing membranous pro f i l e s and tubu l ar a rray s .
3.19
87
I nclu s i on i n c e rebral cor t i c a l n euron showing
f i n gerprint and c rysta lloid pattern s .
3.18
87
88
Part o f ventral horn neuron o f lumbar cord showing a
l a rge numb e r o f inc l u s ion s .
The c ytop l a sm shows
spa r s ity of norma l organe l l e s .
89
F i gure
3 . 20
Page
Membranous inclus ion body in c e l l a t the cort ico­
medu l l ary j unction of the adrenal g l and .
3 . 21
89
Membranous inc l u s ion in a thick bun d l e of a s trocyte
fibre s in the cerebra l cortex of a 25 months old ram
with ceroi d-l ipo fuscino s i s .
3 . 22
91
Pancre a t i c ac inar c e l l showing two type s of
residua l bodi e s .
A curv i l inear body typical for
ovine ceroid-l ipo fu s c ino s i s , and a granul ar , more
e l ec tron den se , body wh i ch may a l so be seen in
norma l sheep .
3 . 23
91
Lymphoid fo l l ic l e in rec ta l wa l l of a 5 months
old ewe l amb .
Ma cropha g e s cont a i n nuc lear remnants
a nd l ipopigment inc l u s i on s
3 . 24
94
.
Ultra s truc tural deta i l o f part o f m a crophage
showing typic a l curvi l i near bod ie s .
Thi n sec tion
obtained from s ame t i ssue a s shown in figure 3 . 2 3 .
3 . 25
94
Granular electron dense inclusion in a m acrophage
in the rectal wall of a 5 months old normal lamb.
These inclusions are encountered in both normal and
affected sheep, and carry no significance in the
diagnosis of ovine ceroid-lipofuscinosis.
95
Page
Fi gure
3 . 24
Autofluore scent ma t e r i a l ou t l i n e s the sweat g l a nds
i n the sk i n of a 25 months o l d ram affected w i th
ceroid-l i pofusc ino s i s .
Before the on set of c l i n i ca l
s i gn s , th i s autofl uore scence i s a l re a dy pre sent i n
affected l amb s a t 4
4.1.
-
5 months o f a ge .
Fam i l y tree of affected i n d i vi dua l s .
95
On ly ma t i n g s
o f heterozygot e s a n d othe r pertinent i ndividua l s
a re shown .
106