close

Вход

Забыли?

вход по аккаунту

?

The oxydase reactions as applied to the megakaryocyte and blood platelet of the rat.

код для вставкиСкачать
THE OXYDASE RESCTIONS A S APPLIED T O
THE &fEGBKARYOCYTE AND BLOOD
PLATEETAET O F THE RAT
CHRISTIANNA S M I T I I , AZARGSRET ROBINSON AND REBECCA T YSON
T h e Walter H . Nerriam Laboratory, Ikpartment of %~Olog?j,
Xoiint HolyoXe College, South Hadlq4, 3fassnckv.sPtts
Whether blood platelets originate from lympliocytes, monocytes, granular leucocytes or cells of the reticulo-endothelial
system as well as from mcgalraryocytes is still an open question. The observations of Wright first made in 1906 and
published again in 1910 regarding the latter as progenitors of
platelets have been many times corroborated. The azurophilic
granules of both platelets arid megakaryocytes as described
by him have been the structural characteristics most often emphasized. Further investigation of the granules has brought
little positive knowledge as to their nature. As is well known
they stain supravitally with neutral red and brilliant crcsyl
blue. Barta ('32) finds that after methyl alcohol fixation the
granules are unstained in 1% eosin, erythrosiri o r acid violet
but that they are colored in Loeffler's methylene blue, methyl
violet and toluidin blue. Roskin and Grinbaum ('26) aiid
Reams and King ('33) report that they do not react to Feulgen's nuclear stain, aiid Corradetti ('34) finds that they are
the only structures of the blood cells to withstand treatment
with hydrochloric acid. IGngsley ( '35, '37) has recently described them as specific granules by which he is able to differentiate even the early stages of the megakaryocytes from
surrounding cells.
Appreciative acknowledgmmt is made to E. Quainine and J. Hastings for
certain technical details used in the various methods.
139
T H E ANATOMIOAL RECORD. VOL. 70, NO. 2 AND SUPPLEMENT NO. 2
JANU.4R.Y.
1938
140
C. SMITH, M. ROBINSON AND R. TYSON
In the present study it was found that the azurophilic granules of both platelets and megakaryocytes were easily and
clearly demonstrated with a combination of Wright's and
Giemsa's stains using Isaac's technique for the marrow. The
granules were uniformly negative in the oxydase and peroxydase tests. This last is contrary to findings reported jn 1935
by Smith and Hastings.
METHODS
After comparing ordinary smears of bone marrow on slides
with others of cells suspended in plasma according to Isaacs
('28, '30) it was decided to use the latter method. By this
procedure all of tlie cells of the marrow were better preserved
and more deeply stained. And what was important for this
study, it was possible by this method t o have both mcgakaryouytes and platelets in llie same medium giving a chance for
more direct comparison of staining reactions. It was found
that the most even and best results were obtained after the
suspension had stood 20 minutes. As a further precaution
the smears of bone marrow suspensions and peripheral blood
were made on two halves of the same slide. Platelets were
also studied in smears of human blood. In the examination
of platelets in blood smears care was taken to make dccisioiis
only on those which could be clearly seen.
All glassware was cleaned according t o the methods ontlined for supravital technique (Sabin, ' 2 3 ) and pipettes and
centrifuge tubes were coated with paraffin. I'Icpauin was used
its the anticoagulant in obtaining tlie plasma. The oxydase
reactions were always started xithin at least 2 hours aftcr
making the preparations.
OHRBRVATIOSS A S D DISCUSSION
The azurophilic granules of the megakaryocytes arid blood
platelets did not react in the oxydase and peroxydase tests
which were used in this study (table 1) although tlie results
were positive with the specific granules of the neutrophiles
and eosiiiophiles and varying numbers of bodies within the
OXYD.4SE REACTIOKS O F MEGAKARYOCYTES
141
monocybes. Whether these latter were exogenous or endogenous could not be determined. An examination of the basophiles was not made. Table 3 shows that while most of the
previous investigators found these granules similarly inactive,
TABLE 1
S?&wkmaryof methods used in t h e slzcdy of the oxydase and perozydase 1.eaclions
of mpgakaryocytes and blood platelets
RRAGENTR
AUTHOP
COUNTEBSTAIN
Block and Perk
Dopa
Rafranin
Imnbright
Blplls naphthol,
hydrogen perodde
Bemidine,
hydrogen peroxide
Cnpper sulphatc,
benzidinc,
hydrogen perosidc
Nitro prnsside,
benzi dinr,
hydrogen peroxide
Methylene blue
McJunkin
Washburxi
(Kolnicr and
Boerner )
TiBLE
COLOR O F OXYJJASE
GRANULES
Dark brown t o
black
Blue to red
Wright 'R
Saf ranin
Blue grern
Wright's
Black
0,
The nrydase uad peroxgdase t r s l s as a p p l i d l o the rr~eyakmyocyleand
blood platelet
METHOD
HCYAN BLOOD
PLATELETS
RAT PERIPHEEAL
BLOOD P U T H L S T B
BAT BONE MARROW
XEGAKARYOCYTES
Block and Peck
Lambright
McJunkin
Sato and Yoshimatsu
Washburn
(Kolmer and
Roerner )
two report positive staining. Contrary to the findings of
Roskin and Grinbaum ('26), with us a successful dopa reaction revealed unstained megakaryocytes surrounded by
strikingly blackened cells of the granular series. Because of
the precipitate the blood platelets were more difficult to see,
142
C. SMITH, &I. ROBINSON AND R. TYSON
but with a safranin counterstain they could bc definitely ascertained. Katsunuma ( ’26) described a positive oxydase reaction in the blood platelets and megakaryocytes which was
difficult to find in normal blood but quite clear in myeloicl
leukaemia, fetal blood and in experimeiital anemia in rabbits.
He believed the oxydase granules corresponded to the azurophilic ones and gave the positive reaction as a microchcinical
support t o Wright’s findings. However, Barta ( ’32) helieved
TABLE 3
Hislory of the oxyduuse and peroxyaase reactions applied l o mcgalaryocytcs and
blood platelets
INVE8TIQATOR
YEAR
G~alinm
1916
1918
Ttiehter
Katsunuma
1923
1926
Eoskin and
Gr inbsum
Knoll
1926
Uarta
1932
METHOD
bAGAAFARYOOYTE9
PLATESETS
Alpha-naphthol,
hydrogen peroxidc
Benzidine,
hydrogen peroxide
MeJunkin
Gierke,
indophenol ox>dasc
Dopa,
Graham henzidine
1927
Blpha-naphthol,
dimethylparaphenylendiamibase
Schulze,
Graham
that the positive oxydase reactioiis of megakaryocytes in infectious diseases were due to phagocytosed material and
therefore not specific differentiations of the cell.
That blood platelets may be derived from cells other than
the megakaryocytes is believed by many, though hard to prove.
I n the study by Birch (’30) on the origin of pseudothrombocytes from leucocytes, she makes the statement that “the
possibility exists that leukoplastids may be normal constituents of the human blood.’’ According to her thesc leukoplastids may be either granular in origin, arising from neutrophiles, eosinophiles, or basophiles, or non-granular, arising
OXYDASE REACTIONS O F MEGAXARPOCYTES
143
from lymphocytes, lymphoblasts, or myeloblasts. Of these
sources only the specific granules of neutrophiles and eosinophilcs were found to have clear-cut peroxydasc reactions in
these studies and table 2 indicates that in normal blood
(human and rat), at least, platelets are not so derived. I n the
peripheral blood of the rat basophiles appear to be absent.
In the case of two neutrophiles in fixed smears, pseudopods
full of oxydase granules were seen and looked as though they
were in the process of being pinched off. No such pieces with
clear oxydase granules were seen free among the cells.
SUMSIARY
1. Tlic azurophilic granules of the mcgakaryocyte stain
morc deeply and easily in cells suspended in plasma (Tsaaes’
method) than in ordinary smears.
2. The aznrophilic granules of normal human blood platelets and of normal rat platelets and megakaryocytes give
negative oxydase and peroxydase reactions.
3. In normal blood there is no evidence that any platelets
arc derived from neutrophiles o r eosiaophiles.
LITERATURE CITED
1932 Uber Xau und Punktion der Megakaryozyten. Folia Haemat.,
vol. 47, p. 168.
BEANS, H. W., AND R. L. EING1933 Feulgen’s ‘nuclealreaktion’ and blood
platelets. Anat. Rec., vol. 55 (suppl.), p. 46.
BIRCH, C. L. 1930 The origin of pseudothrombocytes from leueocytes. Anat.
Rec., vol. 46, p. 45.
BLOCK,
B., AND 8. M. PECK 1930 Der Xachweis der Oxydase i n den Zellen des
myeloisehen Systems durch 3,4-Dioxyphenylalanin. Folia Haemat.,
vol. 41, p. 166.
A. 1934 Ricerche inierocliimiche sui megacariociti e sullc piastrine.
CORRADEWI,
Hsemst., vol. 16, p. 207.
GRAHAM,
G. S. 1916 The oxidizing fcrmeiit of the myelocyte series of cells and
its demonstration b y an alpha naphthol-pyroniii method. J. Ned.
Research, vol. 35, p. 231.
1918-1919 Beiizidine as a peroxidase reagent f o r blood smears and
tissues.. J. Med. Research, vol. 39, p. 15.
ISAAC%,
R. 1925 Alterations of tissue cells in the blood stream. Science, vol.
65, p. 547.
1930 The physiologic histology of the bone marrow. Polia Haemat.,
vol. 40, p. 395.
BAttW, 1.
144
C. SMITH, M. ROBINSOX AND 1L TYSON
KATSUNUMA,S. 1926 Genese der Blut-plLttehen im Lichte der Oxydssereaktiou.
Folia Haemat., vol. 34, p. 29.
KINGSLEY,D. M. 1935 The development of the megakaryocyte i n the pig.
Anat. Bee., vol. 6 1 (suppl.), p. 29.
1937 A technic f o r megakaryocgtes and platelets in sections. Folia
IIacmat., vol. 57, p. 87.
KNOLL,1%'.
1927 Elut urid blutbildende Organc menschliclier Embryonen. Denkschriften der schweixcrischen naturforscliendeii Gescllschaft. M B m . de
la SOC.Helvhtique dea Sci. Nat., vols. 62-64, p. 1.
1931 Approved laboratory technique, p. 89.
KOLNER,J. A., AND F. BCYEENEE
D. Appleton and Co., S e w York.
LAMBRIGEIT,
G. L. 1921 Leukemia: type diagnosis by oxydase method of blood
staining. Am. J. Med. Sci., rol. 161, p. 209.
MCJUNKIN, F. A. 1020 ,4 beiiziciin-polScliroriie stain for blood. J. Am. Xed.
Assoe., vol. 74, p. 17.
RICHTER,Jsf. N. 1925 Leukemia. The relative talucs of cell morphology and
diagnostic aids. Arch. I n t . Med., vol. 36,
thc peroxydasc reaction
p. 13.
ROSKIN,G. O., AND F. T. (+RllvBAUM 1926 A contribution t o the knowledge of
blood platvlrts. Arb. hlicrohiol. Inst. (Mosrow), vol. 2, p. 1 3 7 . Riol,
Abstracts, vol. 2, p. 1577.
RUIN, .'E 1923 Stndies o f living human blood-cells. J o h n s Hopkiiis Hosp.
Bull., vol. 34, p. 277.
SATO,A., AND S. Y o s ~ i n r a ~ s u192.5 Thr perosidnse reirtion in epidemic encephalitis. Am. J. Dis. Child., vol. 29, p. 301.
SMITH,C., AND J. H a s n l r ~ s 1933 A study of the mcgakaryocyte and blood
platelet in the rat. Anat. Rcc., ool. 64 (suppl. no. l), p. 95.
WRIGIIT,J. 11. 1910 The histogenesis of the blood platelets. J. Illorph., vol.
21, p. 263.
Документ
Категория
Без категории
Просмотров
1
Размер файла
287 Кб
Теги
oxydase, platelet, reaction, applied, megakaryocytic, rat, blood
1/--страниц
Пожаловаться на содержимое документа