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His toc hem ie 29 , 355- -361 (1972)

9 by Spr inger -V er l ag 1972

Histochemica l D em onstrat ion o f Succ in ic Dehydrogenase

Act iv ity during Sequent ia l M olar D evelop m ent

in the Sw iss Albino M ouse

D . V i n c e n t P r o v e n z a , Y u n g - F e n g C h a n g , L e sl ie 1 . G a r t n e r , a n d R i c h a r d M . G a r l i t z

D e p a r t m e n t o f A n a t o m y , S c ho o l o f D e n t i s t r y

U n i v e r s i t y o f M a r y l a n d , B a l t i m o r e , M a r y l a n d , U . S . A .

Rec e ived Decem ber 22, 1971

Summary A p p l y i n g t h e d i t e t r a z o l i u m s a l t ( N i tr o - B T ) m e t h o d f o r s u cc i n ic d e h y d r o -

g e n a s e , m u r i n e m o l a r t e e t h w e r e s t u d i e d s e q u e n t i a l l y f r o m t h e c a p s t a g e o f d e v e l o p m e n t

t h r o u g h t h e a p p o s i t i o n a l s t a g e s o f o d o n to g e n e s i s . R e a c t i o n - d i s t r i b u t i o n a n d i n t e n s i t y v a r i e d

r e l a t i v e t o t h e d e v e l o p m e n t a l s t a g e a s w e l l a s t h e z o n e o f m a t u r a t i o n w i t h i n a g i v e n s t a g e .

T h e p e r i p h e r a l c e l ls o f t h e p a r e n t d e n t a l l a m i n a e x h i b i t e d s o m e a c t i v i t y , a s d i d t h e o u t e r

e n a m e l e p i t h e l i u m o f t h e b e l l s t a g e . D u r i n g t h e p e r i o d o f m a t r i x a p p o s i t i o n , c o m p o n e n t s o f

t h e s t r a t u m i n t e r m e d i u m , a m e l o b l a s t ic zo n e a n d o d o n t e b l a s t i c l a y e r r e g io n o f t h e d e n t a l p a -

p i l l a d e m o n s t r a t e d i n t e n s e e n z y m a t i c a c t i v i t y . C e l ls a c t i v e l y e n g a g e d i n e n a m e l m a t r i x p r o -

d u c t i o n d e m o n s t r a t e d a c t i v i t y i n t h e b a s a l a n d d i s t a l c e ll s e g m e n t s . H i g h a c t i v i t y c o n t i n u e d

i n t h e p a p i l l a r y l a y e r o f t h e e n a m e l o r g a n , a s w e l l a s i n t h e c e l ls o f t h e d e n t a l s a c d u r i n g t h e

p o s t a m e l o g e n i c p e r i o d .

n t r o d u c t i o n

L o c a l i z a t i o n o f s u c c in i c d e h y d r o g e n a s e ( S D H ) a c t i v i t y h a s b e e n r e p o r t e d i n

o d o n t o g e n i c t i s s u e o f r o d e n t s ( B u r s t o n e , 1 9 6 0 ; S w a r t z a n d Y e a g e r , 1 9 61 ; B a l o g h ,

1 9 63 ; F u l l m e r , 1 9 6 3, 1 9 6 4 ; R e i t h a n d Z u s s m a n , 1 9 6 4; G o g g i n s a n d F u l l m e r , 1 9 66 )

a n d i n c a n i n e t o o t h g e r m ( M o ri , M i z u s h i m a a n d O s a n a i , 1 9 61 ). T h e s e s t u d i e s

h a v e i n v o l v e d t h e u s e o f a d i t e t r a z o l i u m s a l t ( N i t ro - B T ) i n a p h o s p h a t e b u f f e r e d

s o d i u m s u c c i n a t e m e d i u m , w h e r e t h e N i t r o - B T s e r v e s a s t h e f in a l e l e c t r o n a c c e p t o r

a n d i t s r e d u c t i o n i s v i s u a l i z e d a s d e p o s i t s o f b l u e d i n i t r o f o r m a z a n . R e c e n t l y ,

K e r p e l - F r o n i u s a n d H a j o s (1 96 8) d e v e l o p e d a m e t h o d o f e m p l o y i n g f e r r i c y a n i d e

a s t h e r e d u c i n g a g e n t .

T h e s t u d i e s c i t e d a b o v e w e r e c o n c e r n e d w i t h s p e c i fi c s t a g e s o f o d o n t o g e n e s i s

a n d n o n e i n c l u d e d m o u s e m o l a r s . T h e p r e s e n t s t u d y p r o p o s e s to e l u c id a t e th e

l o c a l i z a t io n o f s u c c in i c d e h y d r o g e n a s e a c t i v i t y t h r o u g h o u t s e q u e n t i a l m o l a r d e v e -

l o p m e n t i n th e S w i ss a l b i n o m o u s e . I t f u r t h e r s e e k s t o p r o v i d e a c o m p r e h e n s i v e

h i s t o r y o f e n z y m e a c t i v i t y p r e p a r a t o r y t o a n e l e c t r o n m i c ro s c o p e c y t o c h e m i c a l

s t u d y .

M a t e r i a ls a n d M e t h o d s

R a n d o m l y b r e d S w is s a l b i n o m i c e o f n o n s p e c if i c s t r a i n , r a n g i n g i n a g e f r o m 1 5 d a y s p r e n a -

t a l t o 8 d a y s p o s t n a t a l , w e r e e m p l o y e d i n t h i s s t u d y . T h e p r e g n a n t f e m a l e s w e re e t h e r iz e d ,

t h e u t e r i n e h o r n s w e r e o p e n e d a n d t h e e x c i s ed f e t u s e s w e re d e c a p i t a t e d . W h o l e h e a d s w e r e

t r e a t e d i n d i v i d u a l l y . I n p o s t n a t a l a n i m a l s , t h e p u p s w e r e e th e r i z e d , a n d t h e m a n d i b l e s w e r e

* S u p p o r t e d b y P H S G r a n t N o . 2 80 0-0 2, N a t i o n a l I n s t i t u t e o f D e n t a l R e s e a r c h , N a t i o n a l

I n s t i t u t e s o f H e a l t h .

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356 D.V. Provenza, Yung-Feng Chang, L. P. Gartner, and 1~. M. Garli tz:

removed. The specimens studied were immediately frozen in liquid nitrogen and oriented on

a microtome chuck to obtain buccolingual sections of developing molars. Frozen serial sections,

14-16 ~z thick, were placed on untreated glass slides and allowed to dry.

The sections were incubated for 10 minutes at 37 ~ C in substrate and phosphate buffered

sodium succinate, pH 7.6 to which an equal volume of an aqueous solution containing Nitro-

BT was added (Nachlas, Tsou, Souza, Chang and Seligman, 1957). Sections were then washed

in physiological saline, fixed in 10 neutra l buffered formalin (10 minutes) , dehydrated in

a graded series of alcohols, rinsed in xylene, and fina lly mounted with Permount.

Control sections were incubated in a substrate-free medium or dipped in the experimental

medium for five seconds. They were then trea ted as described above. Safranin-0 did no t

significantlyenhance visualization, thus its use was omitted.

e s u l t s

The developmental stages invest igated ranged from the cap through the

appositi onal stage,

The basal layer of the presumptive oral epithelium and the parent lamina

of the cap stage demo nst rate d a ve ry slight degree of succinic dehydrogenase

activit y, while the to oth germ p e r s e showed no activi ty.

In the ear ly developing enamel organ the outer enamel epithelium exhibited

moderate acti vity which conti nued into the peripheral cells of the de ntal la mina

an d basal cells of the oral epi thel ium (Fig. 1). The com pon ent s of the stellate reti-

culum, strat um intermediu m, the inner enamel epithelium, dental sac and papil la

were unreactive. Activity was greatest in the cells of the outer enamel epithelinm

located at the apex of the future enamel organ. From this area activity gradually

dimi nis hed along the slope, with its n adi r at t he cervical loop (Fig. 1).

Wi th init iati on of apposi tional stage the cells of the basal layers of the oral

epithelium, as well as those of the pe riphery of the par ent lam ina were unreactive.

The remnants of the outer enamal epithelium, which formed islands and strands

of cells showed no activity. The previously unreactive cells of the stratum inter-

medium and ameloblasts, however , contained reaction deposits . The intensi ty

Fig. 1. Photomicrograph of the early bell stage of odontogenesis of a mouse embryo on the

17th day postconception. Note the enzyme activity (arrows) in the basal layer of the presump-

tive oral epithelium, the peripheral cells of the parent lamina, and in the outer enamel

epithelium. Original magnif ication x 16

Fig. 2. Section of early appositional stage of molar development from a one day old mouse.

Greatest act ivi ty is seen in the trough (T) and crest (C) areas of the enamel organ and decreases

along the slope. Note that the odonteblastic region (arrow) also demonstrates enzyme activity.

Dental papilla (D), ameloblast (A). Original magnification x 40

Fig. 3. High power photomicrograph of the ameloblastic region of a two day old mouse. The

activity is localized in the stratum intermedium S I ) and the infranuclear region of the

ameloblasts (A). Original magnifications x 160

Fig. 4. Appositional stage in a three day old mouse. Note that the highest activities are in

the crest (C) and trough (T) regions of the tooth germ and lowest at the cervical loop (arrow).

Original magnification x 16

Fig. 5. High power photomicrograph of the trough area of Fig. 4. Note activity in the distal

segments (arrows) of the ameloblasts. Original magnificat ion X 160

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Succin ic Dehydro genase Ac tiv i ty in Molar Dev elopm ent 357

Fig . 6 . High power photomicrograph of the cres t a rea of Fig . 4 . React ion deposi ts a re present

in the d is ta l ce l l segments of the ameloblas ts (A) and in ce l l components of , and external to ,

t h e s t r a tu m i n t e r m e d iu m

SI).

Original magnif ica t ion 160

Fig . 7 . Slope area of the molar tooth germ of a s ix day o ld mouse . Note the in tensi ty of the

act i v i ty a long the s lope (arrows) . Minimal ac t iv i ty can be noted in Hertw ig s epi the l ia l root

sheath (H). Orig inal magnif ica t ion 40

Fig . 8 . Presumptive cres t and t rough area of the tooth germ of Fig . 7 . Note ac t iv i ty (arrows)

in the papi l la ry pegs formed by the s t ra tum in termedium. Orig inal magnif ica t ion 40

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358 D .V . Provenza, u Chang, L. P. Gartner, and R. M. Garlitz:

o f t h e r e a c t i o n v a r i e d w i t h t h e t o p o g r a p h i c a l l o c a t i o n o f t h e c e ll . T h e c e l ls o f

f u t u r e c u s p a l a n d t r o u g h a r e a s w e r e m o s t r e a c t iv e , w h i l e th o s e o f t h e s lo p e g ra -

d u a l l y d i m i n i s h e d i n a c t i v i t y . T h e l o w e s t le v e l w a s n o t e d i n th e c e r v i c a l l o o p . T h e

o d o n t o b l a s t i c z o n e p a r a l le l e d t h e p a t t e r n o f t h e a m e l o b l a s ti c a c t i v i t y r e g a r d in g

d i s t r i b u t i o n , h o w e v e r i t w a s c o n s i d e r a b l y w e a k e r ( F ig . 2 ) .

B y t h e s e c o n d d a y o f p o s t n a t a l l if e t h e a m o u n t o f h a r d t is s u e ( e n a m e l a n d

d e n t i n ) p r o d u c e d w a s a p p r e c i ab l e . S u c c in i c d e h y d r o g e n a s e a c t i v i t y i n b o t h s t ra -

t u m i n t e r m e d i n m a n d t h e a m e l o b l a s t s w a s c o n s i d e r a b l e . T h e r e a c t i o n d e p o s i t s

i n t h e a m e l o g e n i c c e ll s w e r e c o n f i n e d in t h e i n f r a n u c l e a r r e g i o n . T h o s e c e ll s w h i c h

w e r e in t h e i m m e d i a t e v i c i n i t y of t h e s t r a t u m i n t e r m e d i u m o f t h e p r o s p e c t i v e

c u s p a l a n d t r o u g h a r e a s a ls o c o n t a i n e d r e a c t i o n p r o d u c t s ( F ig . 3 ). C o n c u r r e n t l y ,

t h e d e n t a l p a p i l l a a n d e s p e c ia l ly th e o d o n t o b l a s t i c r e g io n , e x h i b i t e d g r e a t e r

a c t i v i t y .

I n b o t h o d o n t o b l a s t i c a n d a m e l o b l a s ti c re g i o n s t h e i n t e n s i t y w a s s u s t a i n e d

a t t h e s u m m i t s o f t h e e u s p a l a n d t r o u g h a r e a s, b u t t h e o p e r a t i o n a l t e r r i t o r y o f

t h e e n z y m e w a s s p r e a d o v e r a b r o a d e r , m o r e c e r v i c a l ly l o c a t e d f r o n t .

T h e e n z y m e a c t i v i ty i n c re a s ed b y t h e t h i r d d a y o f t h e p o s t n a t a l l if e b o t h i n

t h e a m e l o b l a s t s a n d s t r a t u m i n t e r m e d i u m . T h e d e n t a l p a p i l l a , a n d e s p e c i a l l y

i t s o d o n t o b l a s t i c l a y e r s h o w e d s t r o n g s u c c i n i c d e h y d r o g e n a s e a c t i v i t y . T h e

g r a d e d l e ve l o f d i f o r m a z a n d e p o s it s , w h i c h w a s g r e a t e s t a t t h e c u s p a l a n d t h r o u g h

r e g i o n s w i t h a n a d i r a t t h e c e r v i c a l l o o p , p e r s i s t e d t h r o u g h t h i s s t a g e ( F ig . 4 ) .

T h i s p e r i o d o f p o s t e m b r y o n i c d e v e l o p m e n t r e v e a l e d t h a t s u c c i n i c d e h y d r o g e n a s e

a c t i v i t y w a s a l t e r e d i n l o c a t i o n , s p e c i f i ca l l y i t a l so a p p e a r e d i n t h e d i s t a l c e ll

s e g m e n t o f t h e a m c l o b l as t . T h e m o s t a c t i v e r e g io n o f t h e a m e l o b l a st , h o w e v e r ,

r e m a i n e d i n f r a n u c l e a r ( Fi g. 5 ). A n i n c re a s e o f t h e r e a c t i o n p r o d u c t w a s n o t e d i n

t h e c o m p o n e n t s e x t e r n a l t o t h e s t r a t u m i n t e r m e d i u m ( F ig . 6 ). T h e p r e s e n c e o f

t h e e n z y m e w a s n o t e d o n l y i n t h e c o m p o n e n t s o f t h e p r o s p e c t i v e d e n ta l s a c i m m e -

d i a t e l y a d j a c e n t t o t h e m o s t a c t iv e r eg i o n s o f t h e s t r a t u m i n t e r m e d i u m .

W i t h f u r t h e r d e v e l o p m e n t , e n z y m e a c t i v i t y r e m a i n e d c o n s t a n t i n d e v e lo p i n g

t o o t h g e r m . H e r t w i g s e p i th e l ia l r o o t s h e a t h ( f o rm e r c e r v i ca l lo o p ) e x h i b i t i e d

l e a s t a c t i v i t y .

B y t h e s i x t h d a y o f p o s t n a t a l d e v e l o p m e n t , s i g n i f ic a n t d i ff e re n c e s i n th e

i n t e n s i t y o f e n z y m e a c t i v i t y b e t w e e n t h e o c c lu s a l s u r f ac e a n d t h e s lo p e s c e r vi c al -

w a r d w e r e n o t o b s e r v e d ( F ig . 7 ). E n a m e l a n d t h e d e n t i n m a t r i c e s w e r e f r e e o f

a c t i v i t y . A c t i v i t y i n t h e d e n t a l s a c w a s m a x i m a l b y t h e a p p o s i t i o n a l s ta g e o f

d e v e l o p m e n t as w a s t h e c a se re g a r d in g t h e r e m n a n t o f t h e s t r a t u m i n t e r m e d i u m

w h i c h w e r e t h r o w n i n t o f o ld s , o r p a p i l l a r y p e g s ( F i g . 8 ).

i s c u s s i o n

S u c c in i c d e h y d r o g e n a s e , o n e o f t h e e n z y m e s o f th e c i tr ic a c i d c y c l e , w a s f o u n d

t o b e v e r y s t r o n g l y i n c o r p o r a t e d i n t o t h e f i n e s t r u c t u r e o f t h e m i t o c h o n d r i a l

m e m b r a n e ( P e t t e , K l i n g e n b e r g a n d B u c h e r , 1 96 2). T h e p r e s e n c e o f s u c c in i c d e -

h y d r o g e n a s e a c t i v i t y i m p l i c a t e s a n a c t i v e m e t a b o l i c s t a te .

S t u d i e s o f s u c ci n ic d e h y d r o g e n a s e a c t i v i t y i n o d o n t o g e n e s is h a v e b e e n l i m i t e d

t o i n d i v i d u a l d e v e l o p m e n t a l s t a g e s a n d s p e c if i c t e e t h ( m o l a r s o r i n c is o r s) . B u r -

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Succinic Dehydrogenase A ctivity in M olar Development 359

s t o n e 1 96 0) i n v e s t i g a t e d h a m s t e r m o l a r s p r o b a b l y in t h e b e l l s t a g e o f o d o n t o -

genes i s . Mor i e t a l . 1 9 6 1) s t u d i e d r a t , g u i n e a p i g a n d d o g m o l a r s i n t h e b e l l a n d

e a r l y a p p o s i ti o n a l s ta g e s, S w a r t z a n d Y a e g e r 1 96 1) i n v e s t i g a t e d m o u s e m o l a r s

i n t h e a p p o s i t i o n a l s t a g e w h i l e B a l o g h 1 96 3) e m p l o y e d g r o w i n g i n c is o r s o f m i c e .

F u l l m e r 1 96 3) a n d R e i t h a n d Z u s s m a n 1 9 64 ) u t i li z e d t h e e n a m e l o r g a n o f r a t

i nc is o rs . T h e s t u d y o f R e i t h a n d Z u s s m a n 1 96 4) w a s c o n c e r n e d o n l y w i t h t h e

a m e l o b l a s ts . G o g g i n s a n d F u l l m e r 1 9 66 ) r e s t r i c te d th e i r s t o d e n t a l p u l p s o f

y o u n g r a t s .

T h e d a t a c o n c e r n i n g s u c c i n i c d e h y d r o g e n a s e a r e i n a c c o r d r e l a t i v e t o i t s

p r e s e n c e i n t h e s t r a t u m i n t c r m e d i u m . E x c e p t f o r B u r s t o n e 1 9 60 ), a ll a r e i n

a g r e e m e n t t h a t t h e e n z y m e is p r e s e n t i n a m e l o b la s t s. M o r i

e t a l .

1 9 6 ] ) a n d

F u l l m e r 1 9 63 ) h a v e d e m o n s t r a t e d t h a t w i t h d i f f e r e n t ia t i o n o f t h e a m e l o b l a s ts ,

a c t i v i t y i s i n t e n s i f i e d . T h e y a d d i t i o n a l l y h a v e i n d i c a t e d p o l a r i z a t i o n o f a c t i v i t y

i n t h e a m e l o g e n ic l a y e r. O f t h e s t u d i e s c o n d u c t e d , o n l y o n e h a s s h o w n t h a t t h e

e n z y m e is p r e s e n t in t h e o u t e r e n a m e l e p i t h e l i u m M o r i

e t a l .

1961) . I n t he s t e l l a t e

r e t i c u l u m , B u r s t o n e 1 9 6 0) , M o r i

e t a l .

1 96 1) a n d F u l l m e r 1 96 3) r e p o r t e d s l ig h t

a c t i v i t y . A l l i n v e s t i g a t io n s h a v e s h o w n s u c c in i c d e h y d r o g e n a s e t o b e p r e s e n t i n

t h e o d o n t o b l a s t s a n d d e n t a l p ap i ll a e. R e i t h a n d Z u s s m a n 1 96 4) n o t e d t h e p r e -

s e n c e o f t h e e n z y m e i n a ll st a g es s t u d i e d a p p o s i ti o n a l t h r o u g h p i g m e n t a t i o n ) .

P o l a r i z a t i o n w a s n o t e d , h o w e v e r , w i t h b a s a l a c c u m u l a t i o n s i n t h e a p p o s i t i o n a l

a n d p i g m e n t s t a g e s . M o r e i n t e n s e a c t i v i t y w a s s e e n i n t h e d i s t a l c e l l s e g m e n t o f

t h e m o r e g i n g i v a l l y lo c a t e d a m c l o b l as t s .

T h e r e s u l t s o f t h e p r e s e n t s t u d y d e m o n s t r a t e t h e d i s t r ib u t i o n o f s u c ci n ic

d e h y d r o g e n a s e t h r o u g h o u t t h e s e q u en c e o f m o l a r t o o t h d e v e l o p m e n t in m ic e .

T h e s e d a t a e s t ab l is h t h a t t h e e n z y m e i s n o t i n i t ia l l y p r e s e n t i n d e t e c t a b l e a m o u n t s

i n th e d e n t a l l a m i n a , r a t h e r i t a p p e a r s i n t h e c a p a n d d i s a p p e a r s in t h e b e ll s ta g e .

I t w a s f o u n d , h o w e v e r , s e le c t iv e l y t h r o u g h t h e e n a m e l o r g a n a n d g e n e r a l ly th r o u g h

t h e r e d u c e d e n a m e l e p i t h e l iu m i n v a r y i n g d e g r e e s o f i n t e n s i ty . T h e b u d s i m i la r ly

e x h i b i t e d n o a c t i v i t y . W i t h h i s t o d i f f e re n t i a t io n , t h e r e a c t i o n p r o d u c t a p p e a r e d

i n t h e o u t e r e n a m e l e p i t h e l i u m a n d g r a d u a l l y d i m i n i s h e d a s t h e e p i t h e l i u m w a s

r e o r g a n i z e d w i t h s t r a n d s a n d c e ll n i d i. T h e c e l ls o f t h e s t e l l a t e r e t i c u l u m w e r e

c o n s i s te n t ly n o n r e a c t iv e w h i le t h e s t r a t u m i n t e r m e d i u m a n d a m e l o b l a st w e r e

i n c r e a s i n g l y m o r e a c t i v e w i t h t h e a c q u i s i t i o n o f f u n c t i o n a l c o m p e t e n c y . T h e

c o r r e l a t iv e p h e n o m e n o n o f a c t i v i t y a n d d e g r e e o f d i f f e r e n t i a t io n a p p e a r e d t o

b e a f u n c t i o n o f t h e s t a g e o f d e v e l o p m e n t . P o l a r i z a t i o n o f a c t i v i t y o b s e r v e d i n

t h e d e v e l o p i n g a n d f u n c t io n a l a m e l o b l a st w a s in h a r m o n y w i t h t h e d a t a o f M o r i

e t a l 1 9 6 1) a n d F u l l m e r 1 96 3). T h e o d o n t o b l a s t i c r e g io n o f t h e d e n t a l p a p i l l a

f o ll o w e d a s i m i la r p a t h , b u t t h e i n t e n s i t y o f re a c t i o n , a s m e a s u r e d b y t h e d e p o s i ts

o f t h e b l u e d i f o r m a z a n , d i d n o t r e a c h t h e l e v e l a c h i e v e d b y t h e a c t i v e r e g i o n s

o f t h e e n a m e l o r g a n .

T h e p r e s e n t s t u d y d e m o n s t r a t e d t h a t t h e d e v e l o p m e n t a l h i s t o r y o f t h e d e n t a l

a n l a g e is p a ra l l e le d b y f u n c t i o n a l a c t i v i t y o f i ts c o m p o n e n t s a s i n d i c a t e d b y

s u c ci n ic d e h y d r o g e n a s e a c t i v i t y . A c c o r d i n g l y , t h e e n z y m e w a s n o t e d t h r o u g h o u t

t h e a p p o s i t i o n a l s t a g e a m c l o g e n e s is ) o f t h e e n a m e l o r g a n . T h e i n t e n s i t y o f l o ca l i-

z a t i o n v a r i e d i n i n d i v i d u a l c e lls a n d c e ll l a y e r s a n d w a s c o m m e n s u r a t e w i t h

m e t a b o l i c a c t i v i t y . T h i s w a s s i m i l a r l y n o t e d b y M o r i

et a l .

1961) , Ba l ogh 1963)

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360 D.V. Provenza, Yung-Feng Chang, L. P. Gartner, and R. M. Garlitz:

and Reith and Zus sman 1964). Ultra struc tural support for this is indicated by

the polarization of the organelles, particularly the mitochondria in the amelo-

bla sts Lenz, 1957; Ny le n an d Scot t, 1958, 1960; Rei th, 1960, 1961, 1963, 1967,

1970; Pannese , 1962; Kal len bac h, 1966, 1968, 1970). The elegant inve sti gat ion

of Reit h 1970) which characterized the amelobla st ult ras tru ctu ral ly during

variou s stages of dev elo pme nt as well as those of Ka ll enb ac h 1968, 1970) sub-

stantia te the histochemical data of the present st udy regarding distr ibuti on of

mitoc hondri a for the ameloblast. The studies of Pannese 1960) on the stellate

ret iculum and the st ratu m interme dium and Kallenb ach 1968) on the papil lary

layer simila rly suppo rt the findings of this study.

e f e r e n c e s

Balogh, K. : Histochemical study of oxidative enzyme systems in teeth and periodontal tissues.

J. dent. l~es. 42, 1457-1466 1963).

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Fullmer, H. M. : In: Oral Histology Inheritance and Development D. V. Provenza), chapt.

18, Philadelphia: J. B. Lippincott 1964.

Goggins, J. F., Fullmer, H. M. : Dehydrogenase histochemistry or the rat molar pulp. Arch.

oral Biol. 11, 1365-1370 1966).

Kallenbach, E. : Elect ron microscopy of the papillary layer of rat incisor enamel organ during

enamel maturation. J. Ultrastruct. Res. 14, 518-533 1966).

Kallenbach, E. : Fine structure of rat incisor ameloblasts during enamel matura tion. J . Ultra-

struct . Res. 22, 90-119 1968).

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regression stages. J. Ultrastruct . Res. 80, 38-63 1970).

Kerpel -Fronius, S., Hajos, F. : The use of ferr icyanide for light and electron microscopic

demonstration of succinic dehydrogenase activity . Histochemie 14, 343-351 1968).

Lenz, H.: Elektronenmikroskopische Untersuchungen an der Ameloblastenschicht. Dtsch.

zahn~rztl. Z. 12, 1568-1579 1957).

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genase activ ity in developing teeth. Histochemie 2,348-355 1961).

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demonstrat ion of succinic dehydrogenase by the use of a new p-nitrophenyl substitut ed

ditetrazole. J. Histochem. Cytochem. 5,420-436 1957).

Nylen, M., Scott, D.: An electron microscopic study of the early stages of dentinogenesis.

PHS Pub. No. 613, Supt. Documents. Washington, D. C. : U. S. Govt. Prin ting Office 1958.

Nylen, M., Scott, D.: Electron microscopic studies of odontogenesis. J. Indiana dent.

Ass. 39,406-421 1960).

Pannese, E. : Observations on the ultras tructure of the enamel organ. I. Stellate reticulum

and stra tum intermedium. J. Ultrastruct . Res. 4, 372-400 1960).

Pannese, E.: Observations on the ultrast ructure of the enamel organ. II I. Intgrnal and

external enamel epithelia. J. Ultras truc t. Res. 6, 186~204 1962).

Pette, D., Klingenberg, M., Bucher, T.: Comparable and specific proportions in the mito-

chondrial enzyme activity pat tern. Biochem. biophys. Res. Commun. 7, 425-429 1962).

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oral Biol. 2,253-262 1960).

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of enamel. J. biophys, biochem. Cytol. 9, 825-840 1961).

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Reith, E. J. : The early stage of amelogenesis as observed in molar teeth of young rats.

J. Ultrastruc t. l~es. 17, 503-526 1967).

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struct . Res. 30, 111-151 1970).

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matura tion of enamel. Arch. oral Biol. 9, 31-38 1964).

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Res. 40, 1291 1961).

Dr. D. Vincent Provenza

Department of Anatomy

School of Dentistry

Univers ity of Maryland

Baltimore, Maryland 21201, U.S.A.

Yung-Feng Chang

Department of Microbiology

Univers ity of Maryland

School of Dentistry

Baltimore, Maryland 21201, U.S.A.

24 Histochemie,Bd. 29