(1)Copyright is owned by the Author of the thesis. Permission is given for a copy to be downloaded by an individual for the purpose of research and private study only. The thesis may not be reproduced elsewhere without the permission of the Author..
(2) 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%).
(3) to. my. pare n t s ... "Ie hebt. 't good edo"'an "... ..
(4) 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 ..
(5) 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 ..
(6) 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.
(7) 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 ..
(8) 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.
(9) 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.
(10) 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.
(11) 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.
(12) 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.
(13) 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.
(14) 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.
(15) 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.
(16)
(2) 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%).
(3) to. my. pare n t s ... "Ie hebt. 't good edo"'an "... ..
(4) 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 ..
(5) 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 ..
(6) 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.
(7) 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 ..
(8) 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.
(9) 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.
(10) 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.
(11) 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.
(12) 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.
(13) 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.
(14) 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.
(15) 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.
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