Doklady Biological Sciences, Vol. 387, 2002, pp. 505–507. Translated from Doklady Akademii Nauk, Vol.
387, No. 4, 2002, pp. 562–564.
Original Russian Text Copyright © 2002 by Novikov, Troitskaya, Gladysheva, Churakov.
PHYSIOLOGY
Specific Anosmia to Isovaleric Acid
in Laboratory C57BL/6 Mice: Recessive Inheritance
S. N. Novikov, V. T. Troitskaya, O. S. Gladysheva, and G. A. Churakov
Presented by Academician Yu.V. Natochin December 25, 2001
Received August 12, 2002
Specific anosmia, a phenomenon of a strong
decrease in sensitivity to certain odorants, was first
described in humans by the American researcher
Blakeslee [1] at the beginning of the 20th century and
provided the basis for Amoore’s highly original theory
of odor discrimination [2, 3]. Although the theory
seemed promising, the physiological and biochemical
processes underlying the phenomenon itself has not
been studied sufficiently. Adequate experimental mod-
els, primarily genetic ones, are required for further
attack on the problem. Researches have great expecta-
tions for the natural chemical compounds, pheromones,
that are used as stimuli [3]. Isovaleric acid (IVA) is one
of them. In some mammalian species [4], it is one of the
major components of the vaginal excretion with signal-
ing function.
In 1977, when testing nine inbred mouse strains for
relative olfactory sensitivity to amyl acetate, pentadeca-
lactone, and IVA by using the conditioned aversion
technique, a group of researches [5] identified two con-
trasting strains, C57BL/6 and AKR, which differed in
the olfactory threshold for IVA concentrations by one to
two orders of magnitude. At the same time, these two
strains did not differ in their sensitivity to the other two
compounds. Hence, the conclusion was inferred that a
specific anosmia to IVA was characteristic of C57BL/6
mice. Afterwards, using various experimental approaches,
the phenomenon was shown to be caused by abnormal
functioning of the peripheral part of the olfactory system
[6–9].
In this work, the role of the initial link of the olfac-
tory system in the mechanisms of specific anosmia to
IVA was studied in the two mouse strains contrasting
with respect to this trait (AKR and C57BL/6) and in
their hybrids (AKB6F1 and B6AKF1) by recording
electroolfactograms (EOGs) from the olfactory layer
preparation.
Pavlov Institute of Physiology, Russian Academy
of Sciences, nab. Makarova 6, St. Petersburg, 199034 Russia
Nizhni Novgorod State University, Nizhni Novgorod, Russia
0012-4966/02/1112-0505$27.00 © 2002 MAIK “Nauka /Interperiodica”
Sixty-three seven- to eight-month-old mice, both
males and females, were used in this study. EOG was
recorded using chlorine–silver electrodes with agar
passage and a tip diameter of 0.1–0.2 mm [6]. The
olfactory stimulation was induced by 0.1 ml of vapor
above a paper filter on which either IVA or its freshly
prepared solution at a concentration of 10–2, 10–3, 10–4,
or 10–5 M was applied. Isoamyl acetate (IA) at a con-
centration of 10–2, 10–3, 10–4, and 10–5 M and its satu-
rated vapor served as reference stimuli. The EOG was
recorded from an isolated olfactory layer of the bone
helices at t = 19°C. Under these conditions, the EOG
recorded from the preparations displayed an invariant
amplitude throughout the experiment in response to the
standard stimulus. From 5 to 19 areas were tested in
each animal.
In C57BL/6 mice, no response to the IVA concentra-
tions from 10–3 to 10–5 M was observed (Fig. 1a). At the
same time, all AKR mice responded to 10–4 M IVA, and
most preparations responded to the concentration of
10–5 M (Fig. 1b). The two mouse strains studied did not
differ in their responses to low IA concentrations
(103−104 M). Thus, C57BL/6 mice with a compara-
tively normal sensitivity to IA exhibited a considerably
reduced sensitivity to IVA; i.e., they were characterized
by a specific anosmia to IVA [6]. The drop in olfactory
sensitivity (an increase in the threshold of the response
by one to two orders of magnitude) was only observed
in males (Fig. 1a).
In the olfactory-lining preparations from mice of
both strains, the areas with a complete anosmia to IVA
and those displaying a normal response to IA were
present simultaneously; no areas insensitive to IA were
found. The EOG analysis of direct and reciprocal
hybrids showed that the anosmia to IVA was inherited
as a recessive trait (Figs. 1c, 1d).
Thus, the sharp interstrainar differences in the sen-
sitivity to the IVA effect were revealed in the prepara-
tions of the olfactory layer by the electrophysiological
method; the recessive inheritance of this trait was also
shown. This suggests that the specific anosmia to IVA
in male C57BL/6 mice is related to hereditary morpho-
functional characteristics of the peripheral part of the
olfactory system.
506 NOVIKOV et al.

(a) (c)
log X C57BL/6 strain (AKR × C57BL/6) F1 hybrids
4.0
Males Females Males Females
3.5

3.0

2.5

2.0

1.5

1.0

0.5

0
(b) (d)
AKR strain (C57BL/6 × AKR)F1 hybrids
4.0 3.5

3.5 3.0
3.0 2.5
2.5
2.0
2.0
1.5
1.5
1.0
1.0
0.5
0.5

0 SV 10–2 10–3 10–4 10–5 SV 10–2 10–3 10–4 10–5 SV 10–2 10–3 10–4 10–5 SV 10–2 10–3 10–4 10–5
M

Fig 1. The EOG amplitude dependence on the concentrations of isoamyl acetate (IA) and isovaleric acid (IVA) in male and female
C57BL/6 and AKR mice and their hybrids. Abscissa shows the concentrations, M (SV, saturated vapor); ordinate shows the loga-
rithm of the EOG amplitude measured in mV. Dark bars, experiment (IVA); light bars, control (IA).

In the recombinant strains of laboratory mice
obtained by the parental cross C57BL/6 × DBA/2, the
genes responsible for anosmia to IVA were mapped to
chromosomes 4 and 6 and shown to display recessive
inheritance [9]. Note that one of the genes responsible
for spermatogenesis and the high level of sperm abnor-
mality characteristic of animals of the C57 family of
strains was also linked to chromosome 4, and the trait
itself was a recessive one [10]. It should be emphasized
that males from this family display abnormal mating
behavior, have low plasma testosterone level and neu-
roendocrine disturbances [3]. Comparison of these data
brings up the question as to whether the specific
anosmia to IVA in male C57BL/6 mice is related to the
abnormal mating function, and whether the same
genetic basis underlies the anosmia at the peripheral
region of the olfactory system and the neuroendocrine
status of the organism.
Thus, the relationships between the sensitivity to
pheromones and reproduction is closely connected with
the urgent problem of the correlation between physio-
logical mechanisms of olfaction, neuroendocrine sys-
tem, and reproduction in general [11]. In our opinion,
of special interest is the hormonal control of expression
of odorant-binding proteins (OBPs), the key and uni-
versal link in the cascade of the perireceptor events of
the olfaction process in terrestrial vertebrates, which
may be responsible for the primary recognizing of the
odorant [12]. In addition, according to [13], in mouse
cells of the olfactory layer, a significant amount of
mRNA transcribed from the Mup gene complex, which
is well-known to be also located on chromosome 4, is
expressed along with OBP [14].
The above data and the results obtained in this study,
taken together, suggest that the C57BL/6 mouse strain
has a genetic linkage group, which, on the one hand,
has an effect on the characteristics of male reproduc-

DOKLADY BIOLOGICAL SCIENCES Vol. 387 2002

 SPECIFIC ANOSMIA TO ISOVALERIC ACID
tive, neuroendocrine function, and formation of the
spermatozoon head shape [3, 10]. On the other hand,
the same genetic group controls the spectral composi-
tion of isomorphic proteins, such as OBPs and MUPs,
which are presumed to differ significantly in the degree
of the affinity to ligands important for communication
(to IVA, in particular).
To provide the basis for the controlled influence on
the organism [14], detailed analysis of the OBP and
MUP differential binding to a number of physiologi-
cally active compounds is required. Note that the dis-
covery of the membrane receptors with high affinity to
OBPs on the cells of the olfactory layer and the lungs in
cows [15] suggests some selective pathways and spe-
cific mechanisms underlying the directed axonal trans-
port of physiologically active ligands within the OBP
complex and (or) its polypeptide fragments upon
endocytosis. This study provides the basis for develop-
ment of some new approaches to the experimental cor-
rection of olfactory deficiency and the mammalian neu-
roendocrine function.
ACKNOWLEDGMENTS
This work was supported by the State Program
“Frontiers in Genetics” (project no. 2.152) and the Rus-
sian Foundation for Basic Research (project no. 02-04-
49273).
DOKLADY BIOLOGICAL SCIENCES Vol. 387 2002
507
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