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HISTOCHEMICAL LOCALIZATION OF ACID PHOSPHATASE ACTIVITY IN TEETH AFTER DECALCIFICATION WITH EDTA
KROLY BALOGH, JR. J Histochem Cytochem 1965 13: 303 DOI: 10.1177/13.4.303 The online version of this article can be found at: http://jhc.sagepub.com/content/13/4/303.citation

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LETTERS

TO

THE

EDITOR

303

wit

ii

fl-giycerophosphate
fur light nsicroscopy

as

substrate
amid
p

nsiusutes

40 sectiosss

glutaraldehyde-fixed tissue were isscubated misiutes, pmsstfixed ims osmium tetroxide bedded us Epiiis for electroum microscopy. fitsonide siund u-tart uric acid were added
the inscuhmatiosi in.if
I

30 phatase activity was suited imi time less ehectruumsdemise, more coarsely gmausuhmtm early secret immun for 20 grausules, oftens cousceust rated sihuuusg their peniplsamid enscry. The most apical lanuiehla of silmost every Sodium Guilgi apparatus sectimuuned, demise bodies, stmmd to suouse rarely uuf small Guuhgi vesicies silso were st-aiumed (Fig.
of 5 3). These
censtratmumus

f(ir

suiixtumes
rat-iomss

sum timatrespectively.

they

coustaimsed

amid 2 With

cuumcesst

observatiumuss of acid altisougis subject A. \. S. tisey

pnuubsibly phsospimsit iou. see stnmd misc coumhd

iusdicsite i)y time imudicate lien P., 1962). Gmmlgi its

cussfumnc-

lie light microscope discrete gmaumuhsim and hansehlar areas uuf staimming were observed supranuelearly (Figs. 1 amid 2). Maumy grasmuies had coiseeustmatimmuss of neactiuums product at them periphery produmci ung a doughsmsut like appearastce. By carefully focusissg at various levels hue uuf
relatively thick were cells.
mt

apparatus,
imums oms this

I m(ussah cmumpartmnemstalizsit (Ic Heuck, (J.

Fmmm furt

specumlsi-

Nmmvikuutl (Lysosouiues, Cssmmtemumum, M.

ed.
LiltIe,

Browui
of et ai.

& Co., 4,

Buist musm, 1964,

(!/tochenu.

p.

36) amid
654, 10:

Nuuvikofl

frozeum noted
iii

sectiuuuns, approximately smot Activity

lamelham 25

areas umf

Histochenu,

activity sicimsar gramsules Imibited

Jassuany
ims-

1965

HiuloLum East Albert College Bronx,

.1. SoBEL Avsus Einsteinof l!edicine Nen York 10461

Staiusismg time cell

was apex.

observed was

ium zynsogeus tuutahly

by the

i)uuths fluoride electrous

assd microscopic

tartmate. level acid phos-

At

HISTOCHEMICAL TEETH
Several
hisised
(if
0i5

LOCALIZATION AFTER
studies
activity

OF

ACID

PHOSPHATASE WITH EDTA*

weeks with tissue
E1)TA 10:232, a tlmawed,

ACTIVITY

IN

DECALCIFICATION
have
us

histumchuemical
stci(1 phsmmspiuatase

heeus
toumth buds

pub-

various J.

species (Cederberg, Dent, Res. Odont. Muind.

but

the Acta 33:646, 1955;

results Anat. 1954; 24: 265, Naujoks,

have 12: 334,

beems

Thirty mice three ausd timeim mstusdibles cossremoved. The uusflxed

hours Histochem. tioins out were chestus ius i)uffered Cytocheuuu-. cumt at 20 us . cuuverships, 10

umhd were decapitated, i(uwer inscisumrs iiere was (lecalcified fun 24

sit 4#{176} (Baiuugis,1. C .

tmadictory Avery, Trim. Zahn. ammd

1951;
Rass.

Banat-ieni, 1956;

1962).
(-

Frozeun
briefly

secdried sit

36: 315, Kieferheilk. Rev.

Dem,tsch.

cmymmststl

18#{176} ), pint C

Schajuuwicz
Latino-

a-mid

room temperature (22#{176} C). After fixatiuuum fun 1 hour ins cold (4#{176} 10 C) formuml-esiiciumm, I lie seeAuuuericana 3: 213, 1958; Quinstarehli, Arch. I/al. Bioi. Oraie 2: 1, 1961; Pourtois, Arch. Biol. (Liege) tiosms were imscubated fur 30 mimnumtes sit nmmuumss 73: 225, 1962). Burstomse, usinsg substituted usaph- temperature ims umsipist iuml AS-Til phmmmsphmsite with pamarumsausilimi as coimpler (Bstrks-i stusd thsuul AS pimosl)hates (J. Hi.stochem-. Cytochem. 7: hexazumsuium 147, 1959; .-lnn. N. V. Acad. Sci. 85:31, 4 1960), Ansdersoms, J. Histocheuun. Cytocheuuu-. 10: 741, 1962). wsis ah)he to dentousst mat-c activity ins bone amid Occasiousally, nsethyh greems was used sis a unucieam staiss. After nimssiusg, sectionss were dried at nouns deustiss nsatmices, but usot mu cells, fmuizess-dmied of tooth germs of msew-bomss modensts. Withs ausother temperature, nsouunted imu Cammada Iisslssuns amid with time highmt microscope. sintuhtauseuuus azo dye coupling method, however, examiumed heavy dye depumsits were seems diffusely in ums-A hmilliasnt red azo dye was seems at the sites umf emuzynse activity. Time gramnular un somewimat erupted nsolams of rodents (Mon et al,, .4rch. amorphous dye deposits were always ismt msicyt 0Histol. Jap. 20: 513, 1960 and J. Osaka Univ. Dent. plasmic amid showed nsiusinsal evideunce (if diffumsioms. Sch, 1: 67, 1961; Yoshiki, Bull. Tokyo Dent. Coil. Ameloblasts umf thse epithehial loop comul nuinned 3: 14, 1962). Mumme necessthy Mon et al. (Histochemie 2: 427, 1962) decalcified rat teeth with EI)TA marked dye deposits t hroughout heir cyt mplsismsi, t (tetrasodium salt of ethylesiediaminetet macct ic whereas ius the zomse of mat mix fumrmat-iumms acid phumsphatase activity was, fumm the muist pant, supraacid) prior t(u sectionsing, but as ins earlier papers (Figs. 1 amid 2). As annehumblasts mmmatured localizatimus muf emszynie activity was marred by nuclear diffusiumum artifacts, Time presemit study was umsdenand became atrophic, the area ansd imsteunsily umf takems to ideustify more precisely sites acid of esizynste activity gradually decreased. Fi msahhy, tIme phosphatase activity in teeth. suprammuchear zone of acid phosphmatase activity merged issto a baumd dye of depumsit * Suppumnted by grasst NB 04155-03 from theimperceptibly
Csibniusi, Ortop. Traumatol. Natiomual Inmstitutes umf Health, Unit-ed

Public

hlealth

Service,

Bethesda,

Maryiansd.

States iss the processes

tipper amid

malpighsiaus the stmsttuns

layer

of

the

gum. showed

Tomues mmii

isitemnuediuns

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304

LETTERS

TO

THE

EDITOR

I tires
I

are

isosummes.

\%ats(mmm (J.

Biophys.

Biochem.

7: 489, 1960) pumst uslsit ed I limit lysmusumumse-hike grsimntmies sit t lie (listslh emm(i uuf secretuury sunmsehuhmlasts
(ytoi. mmssiy
mit

lie

emmanuel

niatmix

imefore t hms

secretimmun. t hme graunimlsin

0mm cmuun-

tlue

lien

hisisn(I,
umfthese presvumt

Heit

thought dim

teunls Time I, I
.,,

vesicles
fiundiusgs megardimmg

mepresemnl smut
lIme

stbsorhmed pemnsit
possihmle a rmmle si snswer

immaterial.
defummit ive muf 1mm Isis 1 steid

(ummn(htmsiuusn

l)hiumspisat

sise

ims

anielogemmesis.

Aun

.c]

(luiestiumun musty cumme frons expemirmicsmtal eludiung lmueahizat iumss si(i(h of l)isuusl)issit muse
sit time ultrastructursil level.

isumrk ussi(l ivi ty

_.l*J

#{149}1

Fia. 1. Msinked sicid pImumsImissitsis ms(tivitv imn anuseiuiiiisists (A) of dpit iselisil loop sumud ims zouse u_if mmit mix formmsstt immum isndiesite(h is Imy azum dye depuisit s (bisiek mum time nsienuuphsotognstphs). Ost-eumchasts Fna. 2. A(id pisosiuhsatase activity iS demumusism stmimnamsuciessr (vt-uiplslsni uuf sunseluuhilasts (amnumws) iimsimng time liusgimal surface uuf t ise milveolar st rated ims zmmnc (if nsat nix fomnsmitiumum. Tunics processes b(umne sulso show insirked emszymuse suet ivity. St-mmutuns (A) (simnow), eusanuieh (E), simud cells umf strat-unms imuterinntemnmediuumms (SI) stnsd enmmunuel (E) mine unegative. mediumns (SI) show nsm enszynuse sictivity. Oduumntox 190.

I)hasts
timeir
resi(I imusm. Odmuumtmmlmlasls shsmuwed mmsmm(lerate (lye (Iemmt mm

(0)
distal (I)) uuf pulls

reveal

coussiderable
butreactive.
sonte

emmzyme
Tonics Time
emmzynie

stetivity

its

deposits
suet

amm(1 tine
Eunsinmmei
(i5i

cells
stud stsnfaee

m hue I uf
(leun

(lcumtstl
I isu renmsstm

pulp
mmcd

weak
blamsk.

cells

cytoplsisnu, sure smut (P) slsumw

fibmers siund time nmesemncinvnmah suet ivity. X

ivi I v.
(lasts

260.
Wit is I lie
sucid
tim

Ostem resided hum

tine

uf m I Isv

stivemmiar

house
descnil)e(l deesilcificat si(t ivit imums I eelsunique lie meiuimn.

imnlcunsely. mmssimmsnumahisimnissues I regsinded it sts 1) mmssii lie vueil

stmn(h
p. .

acid mm iysi

iiiuumsl)iisit misc msuumnsai cmmzynse cuunnpamt e(i. lvsu

I

is

sunfhiciemnt

phuuusPhatase I its imist

stmm

gemsersully Ismmugls nssemnts

Heuuk

sis Present ((IC I)tsve,

sih-siimned
-

peru melsiled
said

uucimenuiicah (if emnzynse

luuestlizmtt

imn Imem (dl mit Lysosouuues, inn

LiI tunes tie, si mmii isis s
tim

Time
mussy
si(I

remankahmle
be
ivmste to

preservsstiuums
the phospisat muses in

suet ivity
muf E1)TA
tim

de mmmi,

capsuhmility uituo

Csunusenuumn,

Bmuuwmm, Bust

(Wehmer 1961

et

(ii.,

19(i3,
isstvc

Iht rsmst suet

usounes

Helu.

Physioi.

Phauuuiacoi. 19(uS

_t eta

19: 97,

hmecmm (Ies(nihmed

Bid. vesielcs

18: fl91

i mmsinmselmuhmiast
Thnese

( lici

hi, .1. (eli were

Jsimmunsiny 21,

KAutoLY

B.-iL(mam-m, .Jut. Pat/u oiogj School

(xe,uerai

19tu3 ( . (list

mnscnmshmrsimne-buumnded

wit in cumuntcsmts muf vsiussihmiedeumsity tIme sul pnmmeesses

fact

Depa utments of Haruard .lledicai

.llassach use//s

fumusund uncan
(if

miund unesin thue ceuster

I

smsseimmhulsnsl TIne s. (stun Inc simvmts

I issi

si(md pismuspissit ase isist mchsenisicsihly ism m

I

Hospital
Eije an(i ,ufl,

and Eui.
.tlssij/i

.hlassae/u / n/u emUC!/

uls(tt
i

unset/s

siut ivi I v I Isese sanse

denmsummnst usit eul aisum suggests

lnstt

I hsese

St mumc-

Rot

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