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THE term Chromaffin System Э and the nature of the Chromaffin Reaction Э.

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THE TERM “CHROMAFFIN SYSTEM” AND THE
NATURE OF THE “CHROMAFFIN REACTION”
B. F. KINGSBURY
Cornell University Medical College, Ithaca, N . Y .
The general use by morphologists of the terms, chromaffin
system, chromaffin cells, chromaffin tissue, chromaffin reaction,
etc., suggests this critical note as timely.
Kohn (’98) first introduced this term as a name for the cellgroups found by him in the sympathetic system, the medulla
of the suprarenal gland, the carotid gland, etc., which assume a
brown color when treated with chromic acid or dichromates.
Observation of this interesting color reaction is older than Kohn’s
extensive and systematic study, going back, I believe, to Henle
(’65). Stilling (’98) preferred the term “chromophile,” while
Poll (’02) has more recently suggested the term phaeochrome
(brown color reaction with chrome salts).
The query is but natural, therefore, as to the cause and nature
of the chromaffin reaction and the attempt to determine this
was precipitated by the peculiar way the medulla of the suprarenal
gland behaved in different fixing solutions.
Naturally, the effect of reagents on the medulla of this organ
has already received considerable attention. The medulla blackens with osmic acid (Schultze and Rudneff, ’65, Bonnamour,
’05); turns green to blue with iron chlorid (Vulpian, ’56, Fusari,
’91, Giacomini, ’02, Mulon, ’03, Stoerck and Haberer, ’08); reduces gold chlorid (Mayer, ’72, Loewenthal, ’94) stains intravitam
with neutral red (Mayer, ’96); stains with basic stains (many
authors).
12
B. F. KINGSBURY
Whether or not the reaction is associated with, and if so limited
to, granules in the cytoplasm has been a matter of discussion
which does not particularly concern us now. Hultgren and
Anderson ('99) Grynfeltt ('03) Kohn ('00) Wiesel ('02) Poll and
Sommer ('03) find that the reaction takes place in granules;
Rabl ('91) Stoerk and Haberer ('08), on the other hand, found
the chromaffi reaction to be a diffuse one. It seems to have
been shown however,-and this is a more pertinent point,-that
after treatment with potassium dichromate or Muller's fluid,
the reaction is diffuse, while potassium dichromate and formaldehyde limits it t o granules in the cells of the medulla, (Kohn,
'04).
Lastly, Neubauer and Langstein ('02) ,pointed out that adrenalin blackens osmic acid solutions as does the medulla, and Ellio t t and Tuckett ('06) state that the intensity of the chromaffin
reaction corresponds to the amount of adrenalin present.
This brings me to the comparison of the reaction of the "chromaffin cells" of the suprarenal and those of adrenalin. It is a
simple matter to test these reactions as I have done by taking
free hand sections of the living suprarenal gland and placing
them in the reagents in watch glasses, carrying out the corresponding test of adrenalin as is customary in test tubes. The
results are arranged in tabular form below:
REAQ~NT
Potassium dichromate
BECTIONS OF BUPRARINAL
(MEDULLA)
uickly
color
SOLUTION OF ADRENALIN
1:lOoo
assumes brown Precipitate, reddish
brown, formed a t once
Potassium dichromate in Quickly assumes brown Precipitate, reddish
color
brown, formed a t once
alkaline solution
Potassium dichromate and Quickly assumes brown Precipitate, reddish
color
brown, formed a t once
formalin
Potassium dichromate and
acetic acid
Assumption of
color which
quently fades
brown
subse-
Precipitate formed by potassium dichromate, soluble on addition of
acetic acid
THE NATURE OF THE “CHROMAFFIN REACTION”
REAGENT
SECTIONS OF SUPRARENAL
(MEDULLA)
13
0OLUTION OF A D R E N U N
1:1000
Zenker’s fluid
Color variable; usually
present, but disappears Not tested
on standing
Silver nitrate (ammoniacal
solution)
Medulla (and zona reticularis) turns a slaty Grayish-black precipitate
(gray) black
Osmium tetroxid (Osmic
acid)
Medulla turns immediately black; subsequent Black precipitate
blackening of cortcx
Flemming’s fluid. (Osmic
acid, chromic acid and
acet.ic acid)
Medulla assumes a black Color turns t o brownish
color, which subse- red and subsequently
quently fades more or blackens; no definite
precipitate
less completely
Gold chlorid
Medulla takes on a brownish red, becoming more
of a purple on standing
Ferric chlorid
color, turning
Medulla a blue-green; Green
brownish red on heatturns brown on adding
ing or adding ammonia
ammonia
Potassium ferricyanide
Brownish red color appears on warming
Fehling’s solution
dedulla turns immediatelj
Reduction in t h e cold
a brick red; cortex a
(green-yellow-red ppt.)
bluish purple
Turns brownish red with
a tinge of pur>le, which
becomes stronger on
standing
Grecn color, turning reddish brown on heating
Benedict’s solution (ont Medulla turns gray; in. On heating, reduction
(whitish-yellow ppt.)
tensified on heating
fluid formula)
The parallelism shown in the simple if rather crude series of
tests, leaves little doubt that the st’ainingreactions of the suprarenal medulla, with osmic acid, chromic acid, ferric chlorid, etc.,
are due to the adrenalin present although of course there may be
other reducing substances present as well. The so-called chromaffin reaction therefore depends not upon any special affinity
14
B. F. KINGSBURY
of the cells of the suprarenal for chromium salts, but simply and
solely is due to a reduction of the chromate to a chromium trioxid (?) combination by strong reducing substances present. The
grouping of organs or cell groups based upon the occurrence of
reducing reactions has little significance unless it can be shown
that the reducing substances are identical. The animal body is
rich in reducing substances; most cell bodies show a moreorless
well marked “chromaffi reaction,” due to reduction of chromates. The nervous system shows in the assumption of the wellknown brown color when it is “mordanted” in dichromate for
the Weigert hematoxlyin method, a chromaffin reaction”
proceeding more slowly, it is true, due in this case to a reduction
by the lipoids present. The cortex of the suprarenal, to go no
further,-shows, particularly in the eona reticularis, a marked
brown color with chromates which has, however, somewhat different reaction conditions. If it is desirable to have the reduction
of chromates represented in the name of the “system,” phaeochrome seems to the writer preferable to chromaffin or chromophile.
The peculiar behavior of the suprarenal in different fixing fluids
becomes clear in the light of the chemical reactions of adrenalin.’
When placed in Flemming’s fluid, the medulla rapidly turns black,
but on standing, the black coloration disappears, the black being
transferred, so to speak (that is, in a suprarenal whose cortex is
rich in lipoids as in the dog or cat or guinea pig), to the cortex
whose rich lipoid content blackens more slowly, so that after
twenty-four hours, the medulla is grayish white, the cortex black.
In osmic acid solution alone the blackening of the medulla is
persistent, the difference in the Flemming’s fluid reaction being
due undoubtedly to the solvent action of the acetic acid probably
assisted by the presence of the oxidizing chromic acid. In potassium dichromate acetic (Tellyesniecky’s fluid), the brown color
of the medulla largely disappears, which is not the case in neutral dichroqfite solutions or dichromate combined with formalin
(a reducing substance) as in Orth’s fluid. In tissue fixed in Zen(‘
1
Cf. also: Oppenheimer: Handbuch der Biochemie, vol.1, pp.768-9.
THE NATURE OF THE “CHROMAFFIN
REACTION”
15
ker’s fluid the brown color which appears a t first gradually fades,
the color residue depending apparently on the duration of the
fixation. Acid fixing fluids are not adapted for the fixation of the
medulla of the suprarena1,-from this side at least. The mixture
employed by Harvey (’06) for the fixation of gastric glands (equal
parts water, formalin, 3 per cent dichromate and saturated solution of mercuric chlorid), or a formula of similar composition
such as Helly’s fluid recommended by Poll therefore give excellent results with the medulla.
16
B. F. KINGBBURY
BIBLIOGRAPHY
BONNAMOUR,
S. 1905 Thesis. Lyon.
ELLIOTT,
T. R. AND TUCKETT,
I. 1906 Journ. Physiol., Bd 34.
FUSARI,R. I891 Atti d. R. Accad. d. Sc. d. Torino, vol. 26; Arch. itaI. d.
Biol., T. 16.
GIACOMINI,
E. 1902 Siena, 1902
GRYNFELTT,
R. 1903 Bull. Sci. de l a France e t d. la Belgique. T. 38.
HARYEY1906 Amer. Jour. Anat., vol. 6.
HULTGREN,
E. O., A N D ANDERSON,0. A. 1899 Skand. Arch. f. Physiol., Bd.
90.
KOHN,
A. 1898 Uebcr die Nebenniere.
Prag. med. Wochenschr., Jahrg. 23,
no. 17.
I900 Arch. f. mikr. Anat., Bd. 56.
1904 Ergeb. d. Anat. u. Entw., Bd. 12.
LOEWENTHAL,
N. 1894 Internat. Monatschr. f . Anat. u. Physiol., Bd. 11.
MAYER,
S. 1872 Sitaungsber d. K. Akad. d. Wiss., Wien., Rd. 66, 3.
1896 Sitzungsber. d. deutsch. naturw.-med. Vereins d. Bohmen;
Lotus.
MULON,
P. 1903 C. R. SOC. Biol., T. 55, Paris.
NEUBAUER,
0. AND LANGSTEIN,
L. 1902 Verh. d. Ges. deutsch. Naturforscher
u. Aerte. Xarlsbad.
POLL,H. 1905 In 0. Hertwig’s Handbuch der Vcrgleichenden Entwickelungsgeschichte der Wirbletiere. Bd. 3, I, pp. 443.
POLLH. AND SOMMER,
A . 1903 Verhandl. d. Physiol. Gesell. Berlin, Jahrg.
1902-3.
RABL,H. 1891 Arch. f. mikr. Anat., Bd. 38.
SCWULTZE,
M. AND RUDNEBF,
,1?. 1865 Arch. f . Mikr. Anat., Bd. 1.
STILLING,
H. 1898 Dic chromophilen Zellcn und Korpcrchen des Sympathicus.
Anat. Ana., 13d. 15, no. 13, pp. 229-233.
STOERK,
0.AND Y . HABERER,
1%. 1908 Arch. f. mikr. Anat.., Bd. 72.
VTJLPIAN 1896 C. R. Acad. d. Sc., T. 43. Paris.
WIESEL,J. 1902 Anat. Hefte., Bd. 19, H. 63.
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