Download CANADIAN TRANSLATION OF FISHERIES AND AQUATIC

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CANADIAN TRANSLATION OF FISHERIES AND AQUATIC SCIENCES
No.
4607
Sexual dimorphism in the serum proteins of Salmo salar:
study
elctrophi
by A. Drilhon, .and J.M. Fine
Original Title: Dimorphisme sexuel dans les protéines sériques de Salmo
salar: étude électrophorétique
From: C.R. Seances Soc. Biol. Vol.-CLVII: p. 1897, 1963
Department of Fisheries and Oceans
Newfoundland Environment Centre
St. John's, Nfld.
1979
4
pages typescript
2050592
p. 1.1
zli‘o7
Printed with the periodical:
Minutes of the meetinos of the Société de Biologie
Meeting of November 16, 1963 -- Tome CLVII, No. 11, 1963, p. 1897.
Printed in France
Sexual dimorphism in the serum proteins of Salmo salar:
Electrophoretic study
by Andrée Drilhon and Jean M. Fine
A few partial results acquired on the serum proteins of Atlantic Salmon
(Salmo salar) during its various stages of growth
tlidc-urm-¢2
(1*) encouraged us to again ty«
-bop this study at the electrophoretic level and in terms of immunology by preparing
several immune serums.
The success of this study was made possible by a sufficient
number of Salmo salar samples (*) collected from the Frayères du gave d'Oloron
II
spawning grounds in December and at Peyrehorade sur l'Adour in April. We were
able to examine male and female spawners, weighing 3 to 7 kg and measuring 40 to
98 cm, parrs, sexually mature young salmon which have yet to migrate, as well as
"early run" salmot in April, sexually inactive adult salmon, smolts, ànd mendeds
preparing for the catadromous migration.
Physiological and endocrinological
details concerning this species at its various levels
in ethe works of Maurice Fontaine
,t
e-n4-44-is
cf development are contained
(2*).
To date, studies of
serum proteins through electrophoretic methods (electrophoresis on paper) have
.2 ,
been confined to American species etWorhynchus kiisutch) (3*).
(1*)
A. Drilhon, Bull. Inst. Océan., 1960, No. 1168, p. 1.
(*)
These serums were collected and sent to us by M. Fontaine, to whom we
hereby extend our warmest thanks.
i
205059 2
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p. 2.1
.
We studied these various samples
ÿielectrophoresis in gelose gel,
agarose gel and starch gel.
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Fig. 1: Electrophoresis in gelose at pH_8.6 of salmon serums -males (no. 8, 15 and 18), females (no. 11 and 12) and
parrs (no. 1 and 2). Amidoschwarz coloration of proteins.
Males and females are differentiated by a proteinogram
showing the importance and position of globulins near the
serum deposit point.
Techniques -- The Grabar (4*) technique is used to perform electrophoresis in
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gel ose on 13 x 18 cm glass plates, 1,5' gelose ^f^r-ecf^with^^ veronal--4a^s.
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pH =8.6, according to Hirschfeld (5*).
pH:8.6,
(2')
M. Fonlainc, F.therirrlfiff, 19611, t. 1t;, P. 133,
y
(3') 1V. E. 1'anNlunw et Chun;; 1\'ai Un. J_'ishtrirs Research
1Joizrcf of (: rr.mlrl,
f3, p. :1.
(J') 1'. Ginli;ir ct P. Iturtin, Annl.\.o iinniuni.;'Irrtrnphnritiqur, .l(;^.,^^^ et
Cu: ^d., l'aii> IJGP.
(5') I. Il r^ch(cI
.ïrirnrr 7ni./x, 1!>I:11 L. 7, p. 1S.
2050592
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p. 3.1
Electrophoresis in agarose gel is performed in exactly the same manne5
-ertiley..449,4J4, with gelose being replaced by 1.5% agarose. Amidoschwartz coloration
of proteins is carried out as per Uriel and Grabar (6*).
A modified (8*) version of the horizontal method described by Smithies (7*)
is used to perform electrophoresis in starch gel with a discontinuous system of
Orate buffers (9*).
-
Results -a. SEXUAL DIMORPHISM IN SALMON
Figure 1 shows that -Unit,- the proteinogram of the sexually mature female
may be distinguished from that of either the very young (parr) or spawning (adult)
male by means of electrophoresis in gelose.
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11
—
a
Fig. 2: Electrophoresis in starch gel (discontinuous system of triborate
buffers) of male (no. 8 and 15) and female (no. 11, 12 and 13)
salmon serums.
A difference is also observed between male and female proteinograms.
Three principal fractions are readily distinguished, two of which occupy
the same location in all specimens examld, regardless of age or sex: albumin, the
most anodal fraction at pH 8.6; and th 'ajority fraction of the serum, located
,
behind the albumin. Conversely, the third important component occupies either
01', J. Uri e l ,t P, t;r:diar, .1 rn. It,ol .
Pfixtertr, 19f , t. DO, p. 427.
if it.chetn,
1955 • t . 6/ . p . 62D .
J .1. :NI. Fine t .1.
net,.
t. 11, p. 419,
( 9 ',
1).
.V‘thire. 19:■ 7, 1. In, . p. 1172,
( 7') 0.
2050592
p. 4.1
a cathodal position in relation to the serum deposit among all males examined, or an
anodal position 2mong females.
This same distinction also occurs in agarose gel
electrophoresis (**).
A larger number of component; may be distinguished by means of electrophoresis in starch gel. In this medium, proteins of identical electrophoretic
mobility occupy different locations according to their molecular size.
Here again,
male and female proteinograms are differentiated by the existence in females of an additional fraction, the location (lesser mobility than in gelose) of which
ry-Y1
sees to indicate a heavy molecular weight (see figure 2).
b. GROWTH STAGES
There are quantitative variations in the distribution of serum fractions
among adult spàwners, running salmon, parrs, smolts and mendeds.
These differences are particularly visible after electrophoresis in
agarose gel, manifesting themselves in parrs and smolts through a strong elevation
of the area behind the albumin (mobility Q( 1 ).
These quantitative differences
observed in the various growth stages may be related to hormone changes physiologically
uncovered in the works of Fontaine and teloup (10*, 11*, 12*).
Conclusions -- By means of zonal electrophoresis in qelose and starch gel, a
significant difference may be uncovered in the distribution of proteins in
Salmo
salar serum, making possible the distinction of males and females.
(Laboratoire de Physiologie des Etres marins, Institut Océanographique et Laboratoire
d'Immunochimie, t.entre national de la Transfusion sanguine, Paris).
(**) Agarose gel results from a purification of gelose, leading to a reduction
in acid yroups and, consequently, a reduction in the electroendosmotic current
during electrophoresis.
(1 )•) J. Leloup. C. R.
1053, 1. 217, p. 2451.
(11') :\1. Fontaine ct J.
1.cloup. Grn. und
p. :Hi.
compnr. FruincrinnI., 19f;:!.
(1'2') .1. t.tionp, Assou.pj,
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p,
(lt lanzue française, 19F; I, Herne, t.