ISSN 00329452, Journal of Ichthyology, 2011, Vol. 51, No. 6, pp. 474–483. © Pleiades Publishing, Ltd., 2011.

Original Russian Text © A.A. Bolotovskiy, B.A. Levin, 2011, published in Voprosy Ikhtiologii, 2011, Vol. 51, No. 4, pp. 559–568.

Effect of Thyroid Hormones on the Development of Pharyngeal

Dentition in Roach Rutilus rutilus (Cyprinidae, Cypriniformes)
A. A. Bolotovskiya and B. A. Levina, b
a
Papanin Institute for Biology of Inland Waters, Russian Academy of Sciences, Borok, Yaroslavl oblast, 152742 Russia
b
Severtsov Institute of Ecology and Evolution, Russian Academy of Sciences, Leninskii pr. 33, Moscow, 117071 Russia
email: lehabolot@mail.ru
Received March 15, 2010

Abstract—Development of teeth on the pharyngeal bones of roach Rutilus rutilus and the effect of thyroid
hormones on this development are investigated. The addition of exogenous triiodothyronine leads to accel
erated development of the teeth, but the deficit of triiodothyronine (provoked by the addition of thiourea in
the media) stimulates the retardation of this development. Change of developmental rate of the organism
leads to change in the definitive state of the pharyngeal teeth formula. Owing to accelerated development, the
number of teeth significantly decreases, and the formulas 55, 54, and 44 appear instead of the typical for
mula 65 in the control group and in the fish from natural populations. Retarded development of the organ
ism leads to increased frequency of occurrence of the formula 66. The directed asymmetry in the numbers
of pharyngeal teeth with the formula (65), most likely, is connected with different types of teeth development
on the left and right pharyngeal bones.
DOI: 10.1134/S0032945211040035
Keywords: pharyngeal teeth, teeth development, heterochrony, thyroid hormones, goitrogens, Cyprinidae.

In the fishes of the family Cyprinidae, teeth on the
jaws are absent, but the pharyngeal teeth are well
developed on the fifth pair of modified branchial
arches. The pattern of distribution and number of pha
ryngeal teeth are the stable and (often) species specific
characters traditionally used in the systematics of this
large family (Berg, 1912; Vladykov, 1934; Chu, 1935;
Wu et al., 1981; Howes, 1991). Recent experiments on
the retardation and acceleration of development of
African barb Labeobarbus intermedius by means of
manipulation of thyroid status of the fish show that
substantial changes both in comparatively variable
character (scale number) and in the stable characters
(number of pharyngeal teeth and even number of the
rows of the pharyngeal teeth) can be induced (Smirnov
et al., 2006; Smirnov and Levin, 2007; Shkil et al.,
2010).
Only one row of pharyngeal teeth (in comparison
to three rows in African barb of the subfamily Barbi
nae) is observed in roach Rutilus rutilus, a representa
tive of the subfamily Leuciscinae, the family Cyprin
idae. In addition, roach is characterized by directed
asymmetry in the number of pharyngeal teeth: the
majority of fish possess the formula 65. The develop
ment of teeth and histogenesis of pharyngeal teeth of
the roach have been described previously (Grib, 1930;
Cheprakova, 1958). In earlier studies, the develop
ment of teeth is reconstructed based on histological
sections. However, according to Lange (1967), this
method can lead to erroneous conclusions on the
number of rows and the number of main and replaced
teeth. Therefore, a method of reconstruction of the
teeth structure of pharyngeal bones on stained and
cleared preparations of the specimens is applied in this
study.
The goals of this study are the investigation of nor
mal development of teeth on the pharyngeal bones in
roach, including their normal definitive state, as well
as the examination of their development at different
thyroid states induced by deficit or excess of thyroid
hormones.
MATERIAL AND METHODS
Material from this study was represented by the lar
vae and juveniles of roach grown from eggs obtained
from a female and fertilized by sperm of a male using
the “dry” method. The spawners were caught in the
Rybinskoe Reservoir near Borok village, Yaroslavl
oblast. Ten minutes after fertilization, the eggs were
divided into three groups and were placed into differ
ent media: TH group, 1 ng/ml triiodothyronine alka
line solution, Т3; THIO group, 0.02% thiourea solu
tion, a goitrogen that blocked the synthesizing activity
of the thyroid gland; and the control group kept in
dechlorinated tap water. All solutions for the treat
ments were prepared based on tap water. During incu
bation, the eggs were kept in the bowls, and, after
hatching, the larvae were kept in aquariums, in 40 l
volume, with aeration. The cultivation conditions

474
 EFFECT OF THYROID HORMONES 475

ce0 an1
po1
an2
7 A4
6 A5 A3
VII A2
5 an3
iii
V VI
4 A1
IV
3 i ii
II III
2
1 I

Fig. 1. Development and distribution of pharyngeal teeth in Cyprinidae (Nakajima, 1991; modified): 1–7 are teeth generations;
po1, ce0, an1, an2, and an3 are positions of the teeth families; I–VII are teeth of the central (bended) part of the pharyngeal bone;
i–iii are teeth attached to the anterior part of the pharyngeal bone; A1–A5 are teeth of the main row at definitive condition.
Explanations are given in the text.

(temperature, photoperiod, stocking density, and
feeding regime) were identical in all experimental
groups. Approximately half of the water was changed
in the aquariums daily, but the concentrations of the
chemicals in the experimental groups remained the
same. The fish were fed zooplankton and Artemia for
the first 2 months, and then they were fed Biomar
compound feed for the trout.
From the age of 9 days (from the fertilization), the
exemplars (from three to six individuals) were fixed in
4% formaldehyde. During the early development, the
larvae were fixed on each second day, and then the
periodicity of fixations was gradually increased. The
total length (TL) of the fish was measured, and average
values of this parameter (n = 3–6 exemplars) are given
in the text.
Fixed exemplars were stained in alizarin dissolved
in ethanol according to the modified method (Taylor
and van Dyke, 1985). Following staining, the speci
mens were cleaned in water and cleared in alkaline
solution (KOH). For the final clearing, treatment, and
subsequent storage, the exemplars were placed in
100% glycerol. The pharyngeal bones with surround
ing tissues were removed from the cleared preparations
throughout the branchial openings, and they were
analyzed under a Motic DM143FBGG binocular
microscope. The photographs were obtained using the
Leo 1420 electron scanning microscope (Carl Zeiss).
In the three groups, the development of pharyngeal
teeth was studied to the age of 24 days. The border
between the juvenile and definitive states of the char
acter was determined based on the disappearance of
the location of the teeth in several rows and termina
tion of the addition of new anterior teeth. The defini
tive formula of pharyngeal teeth was assessed in 72,
173, and 34 specimens from the control, TH, and
THIO groups, respectively.
The terminology used in this study was developed
by Nakajima (1979, 1984, 1987), who investigated
development of pharyngeal teeth in several species of
cyprinids (Fig. 1). According to Nakajima (1984), the
replacement waves are represented by the series of
replaced teeth of a generation (horizontal dashed lines
1–7); the teeth family (the teeth of different genera
tions connected by a solid line in Fig. 1) is initial tooth
and replacement teeth, which appear at the same posi
tion on the pharyngeal bone). The positions are as fol
lows: po1, ce0, an1, an2, and an3. Capital Roman
numerals (I, II, III, IV, etc.) are used for the designa
tion of the teeth on the central (bended) part of the
pharyngeal bone. Small Roman numerals (i, ii, iii,
etc.) are used for the designation of the teeth devel
oped on the anterior part of the bone. The first tooth
(I) appears in the central position ce0. The anterior
positions of the teeth are designated as an1, an2, and
an3; the posterior positions are designated as po1, po2,
etc.
In the text, the term “pharyngeal tooth” is used
exclusively for the teeth attached to the pharyngeal
bone. The development of the pharyngeal tooth begins
from the formation of tooth anlage developing into the
tooth crown (Grib, 1930). The first structure, which is
observed visually after alizarin staining of the speci
men, is represented by the tooth crown. Thus, the
teeth not attached to the bone we call “crowns.”
The significance of differences in the frequency
distribution of teeth in different groups was assessed
based on the Kolmogorov–Smirnov test.
RESULTS
The sequence of the appearance of the crowns,
their attachment, and resorption of pharyngeal teeth
in roach larvae from the control and experimental
groups are illustrated in Fig. 2. The presence of more

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476 BOLOTOVSKIY, LEVIN

Age Control group TH group THIO group

from
fertili
zation,
days
11 TL 6.4 mm TL 6.1 mm TL 6.5 mm

13 TL 6.4 mm TL 6.3 mm TL 6.6 mm

15 TL 6.6 mm TL 6.5 mm TL 7.4 mm

17 TL 7.6 mm TL 7.3 mm TL 7.8 mm

19 TL 7.7 mm TL 7.6 mm TL 8.0 mm

22 TL 11.3 mm TL 8.8 mm TL 8.1 mm

24 TL 11.6 mm TL 10.4 mm TL 8.6 mm

Fig. 2. Scheme of development of the pharyngeal teeth in roach Rutilus rutilus in the control group and two experimental groups:
(  ) are teeth generations; (䊉) are teeth attached to the pharyngeal bone; (䊊) are teeth crowns not attached to the pharyngeal
bone (appearing recently); ( ) are teeth subjected to resorption.

than a row of pharyngeal teeth is registered in the early
ontogeny of roach in all groups.
Development of Pharyngeal Teeth in the Control Group
Development of pharyngeal teeth in the control
group begins from the appearance of the crown of the
first tooth in the position of ce0. At the same time, the
ossification of the fifth branchial arch has not begun,
which is supported previously by Grib (1930). By the
age of 9 days (6.2 mm TL), calcified crown of the first
pharyngeal tooth I (on the right and left sides) is seen
in the pharyngeal area. By the age of 11 days (6.4 mm
TL), two other crowns (an1 and po1) appear. They are
located anteriorly and posteriorly from the first tooth;
the first tooth is attached to the ossified part of the
pharyngeal bone. The crowns of teeth II and III form
the second replacement wave. By the age of 13 days
(6.4 mm TL), the crown of tooth IV of the third wave
is observed. It is located behind two teeth of the second

JOURNAL OF ICHTHYOLOGY Vol. 51 No. 6 2011

 EFFECT OF THYROID HORMONES
wave opposite tooth I (at the position ce0). At this age,
three teeth are seen on each pharyngeal bone. By the
age of 15 days (6.6 mm TL), the same number of pha
ryngeal teeth is registered, but the crowns of the teeth
V and VI at the positions of an1 and po1 (in addition
to the tooth IV not attaching to the bone) are seen in
the largest specimen (7.5 mm TL). By the age of
17 days (7.6 mm TL), the difference between the right
and left pharyngeal bones in the pattern of teeth
attachment is observed for the first time. On the left
pharyngeal bone, the crown of tooth I at position an2
appears; this tooth is attached to the bone in the largest
(8.5 mm TL) exemplar. Tooth IV is observed on the
bones of both sides. By the age of 19 days (7.7 mm TL),
pharyngeal tooth I at position ce0 is resorbed, and
tooth i is seen on the left pharyngeal bone. By the age
of 22 days (11.3 mm TL), the crowns of teeth VII,
VIII, and IX appear on both pharyngeal bones, and the
crown of tooth iii at position an2 is also seen. At the
same time, the crown of tooth iii substitutes tooth i on the
left bone, and the crown of tooth iii appears for the first
time on the right bone. By the age of 24 days (11.6 mm
TL), resorption of teeth II, III, and IV is observed at
the left side; teeth VII, IX, and iii are attached to the
pharyngeal bone; the crowns of tooth X at the position
ce0 and tooth iv at the position an3 appear. By this age,
teeth II and III are resorbed on the right bone, teeth
VII, IX, and iii are attached to the bone, and the crown
substituting tooth iii at position an2 is observed.
Development of Pharyngeal Teeth in the TH Group
The development of pharyngeal teeth in the
TH group occurs slightly faster than in the control
group (that is clearly seen at earlier developmental
stages). By the age of 9 days (5.9 mm TL), a tooth is
observed on the ossified part of the pharyngeal bone at
position ce0. In addition, two crowns are seen at posi
tions an1 and po1. By the age of 11 days (6.1 mm TL),
three teeth on the bone and the crown of tooth IV at
position ce0 are registered. By the age of 13 days (6.3
mm TL), the crown of tooth i is seen on the left bone,
and the right bone is without changes. By the age of 15
days (6.5 mm TL), tooth IV is attached to the right
bone, and two additional crowns of teeth V and VI at
positions an1 and po1 are registered. By the age of 17
days (7.3 mm TL), tooth i at position an2 is attached
to the left bone. At this stage, the first tooth (I) of the
left bone at position ce0 is resorbed. By the age of 19
days (7.6 mm TL), tooth I is resorbed on the right
bone, teeth V and VI are attached to the bone, and the
crowns of teeth VII and iii are seen. At this stage, the
pattern of teeth development of the left bone is similar
to that at the age of 17 days. By the age of 22 days
(8.8 mm TL), two additional teeth (II and III) are
resorbed on the left bone; teeth V, VI, VII, and iii are
attached to the bone; and the crowns of the teeth VIII,
IX, and iv are seen. On the right bone, tooth II is
resorbed. By the age of 24 days (10.4 mm TL), changes
JOURNAL OF ICHTHYOLOGY Vol. 51 No. 6 2011
477
are registered only on the right bone: tooth III is
resorbed, teeth VII and iii are attached to the bone,
and the crowns of teeth VIII and IX are seen.
Development of Pharyngeal Teeth in the THIO Group
The development of pharyngeal teeth in the THIO
group, in general, is similar to that in the control
group. By the age of 9 days (6.3 mm TL), the crown of
the first tooth at position ce0 is observed. In a speci
men, the crown is seen only on the left pharyngeal
bone, and the crowns are absent in another specimen.
By the age of 11 days (6.5 mm TL), the first tooth is
attached to the bone, and two crowns of teeth II and
III appear. In the larvae at the age of 13 days (6.6 mm
TL), teeth II and III at positions an1 and po1 are
attached to the bone, and the crown of tooth IV sub
stituting the first tooth at position ce0 is registered. By
the age of 15 days (7.4 mm TL), the crown of tooth i at
position an2 is seen on the left pharyngeal bone. At the
same time, tooth IV is attached to the right bone, and
the crowns of teeth V and VI at positions an1 and po1
appear. By the age of 17 days (7.8 mm TL), teeth i at
position an2 and IV are attached to the left bone, and
the crowns of teeth V and VI at positions an1 and po1
appear. At this age, the pattern of distribution of the
teeth on the right bone remains the same. By the age of
19 days (8.0 mm TL), the first tooth is resorbed on
both pharyngeal bones. On the right bone, two crowns
of teeth V and VI are attached to the bone. On the left
bone, only tooth V at the position po1 is attached to
the bone. By the age of 22 days (8.1 mm TL), tooth VI
is attached to the left bone, and the crowns of teeth VII
and iii are seen at positions ce0 and an2, respectively.
At this age, the changes in the position of the teeth on
the right bone are not registered. By the age of 24 days
(8.6 mm TL), tooth VII is attached to the left bone,
and teeth VII and iii, as well as the crowns of teeth VIII
and IX, are seen on the right bone.
Features of Definitive Teeth Distribution
Parents of the experimental fish possess the teeth
formula 65. The type of teeth distribution usual for
adult individuals is registered by the age of 45–50, 40–
45, and 50–55 days in the control, TH, and THIO
groups, respectively. Thus, the development of pha
ryngeal teeth terminates in the TH group earlier than
in the control group, and this development in the
THIO group terminates later than in the control
group.
Photographs of all variants of pharyngeal teeth dis
tribution obtained in the experiment are illustrated in
Fig. 3. Frequency distribution of the formulas of pha
ryngeal teeth in the control group is usual for the spe
cies (table). Asymmetrical teeth distribution (65) is
observed in the majority of exemplars (94.4%), and
remaining individuals possess the formulas 55 (1.4%)
or 66 (4.2%). In the TH group, the formula of pha
478 BOLOTOVSKIY, LEVIN
Distribution of the variants of the formulas of pharyngeal teeth in the control and experimental groups of roach Rutilus rutilus
Group (number of exemplars)
44
Control (n = 72) – –
TH (n = 173) 7 35
THIO (n = 34) – –
ryngeal teeth in the modal group is different from that
in the control group. The typical formula is 55
(74.6%), and the formula 65 is observed only in two
exemplars (1.2%). In addition, the formulas 54 and
44 (20.2 and 4.0%, respectively) are observed in the
TH group; such formulas are not registered either in
the control or THIO groups. Significant differences (p <
0.001) in frequency distribution of the formulas of
pharyngeal teeth are registered between the TH and
control groups. In the THIO and control groups, the
modal formula is 65 and the differences in frequency
distribution of the formulas of pharyngeal teeth are not
significant. However, a trend towards deviation of fre
quency distribution to the formula 66 (in more than
20% of individuals) is registered in the THIO group.
DISCUSSION
Thyroid hormones (TH) are included in the hypo
thalamopituitarythyroid axis (Power et al., 2001;
Blanton and Specker, 2007), and they are important
for many metabolic and morphogenetic processes as
well as for metamorphosis, regulation of reproductive
system and regeneration of different organs (Pickford
and Atz, 1957; Leatherland, 1982; Blanton and
Speckler, 2007; Sekimizu et al., 2007). As is known,
the levels of TH depend on the ontogenetic rate of the
fish (Miwa and Inui, 1987; Brown, 1997; de Jesus
et al., 1998; Okada et al., 2005; Smirnov et al., 2006).
Goitrogens (in our experiment, thiourea) are chemi
cal inhibitors of TH synthesis of the thyroid gland, and
their addition into the medium leads to the deficit of
TH in blood plasm (Brown, 1997). The deficit of thy
roid hormones leads to retardation of the transition
from the larva to juvenile and to late appearance of
scale cover in zebra danio Danio rerio (Brown, 1997),
to inhibition of metamorphosis in bastard halibut
Paralichthys olivaceus (Okada et al., 2005), and to
retardation of development of scale cover and differ
entiation of the fin rays in African barb Labeobarbus
intermedius (Smirnov et al., 2006; Smirnov and Levin,
2007). However, the addition of exogenous TH leads
to earlier differentiation of the pectoral fins and to
faster growth of the pelvic fins in zebra danio (Brown,
1997), orangespotted grouper Epinephelus coioides
(de Jesus et al., 1998), and African barb (Smirnov
et al., 2006).
In the experiment conducted in this study, the
development of the pharyngeal teeth in roach from the
Formula
54 55 65 66
1 68 3
129 2 –
2 25 7
TH group was faster (and in the THIO group, it was
lower) than in the control group (Fig. 2). Thus, the TH
and THIO groups can be designated as the groups with
accelerated and retarded development, respectively
(Levin, 2010). In the group with accelerated ontogeny,
accelerated teeth development was observed mainly at
early developmental stages (age of 9–11 days). On the
contrary, in the group with retarded ontogeny, the rate
of teeth development was similar to that in the control
group to the age of 19 days, but this rate was substan
tially slower than that in the control group afterwards.
The differences in the developmental rate, most likely,
were responsible for directed changes of the definitive
state of the character in the experimental groups. A
similar reaction to the acceleration or retardation of
the ontogenetic rate followed by the manipulation of
the thyroid status of roach was registered for the num
ber of scales in the lateral line (Levin, 2010).
The asynchronous acceleration of development of
the pharyngeal teeth under the influence of TH and
retardation of their development induced by the
goitrogen can be explained in the following way. The
egg envelope (chorion) represents the barrier for the
distribution of exogenous triiodothyronine (Raine
et al., 2004). However, the embryos of zebra fish are
capable of effective consumption of T3 from water
solution by the age of of 8 h from fertilization (Walpita
et al., 2007). The addition of thiourea (the goitrogen)
leads to the decrease or blockage of synthesis of endog
enous thyroid hormones in the thyroid gland; this gland
is developed in zebra danio by the age of 55 h after fer
tilization (Alt et al., 2006). Thus, we can suggest that
the effect of exogenous T3 begins earlier in comparison
to the effect of thyourea.
The primary pharyngeal teeth of roach are distrib
uted in two rows (Vasnetsov, 1939; Cheprakova, 1958;
Lange, 1967). The distribution of pharyngeal teeth in
several rows is found in larvae of all studied species of
the families Cyprinidae, Cobitidae, and Catostomi
dae, and the correlation between the numbers of larval
teeth rows and definitive teeth rows is often absent
(Weisel, 1967; Nakajima, 1979, 1984, 1987, 1990;
Nakajima and Yue, 1989). Distribution of the pharyn
geal teeth in several rows in the larvae is connected
with the presence of teeth from different generations
(“replacement waves”) on the pharyngeal bone. Teeth
of the earlier wave are not resorbed, but teeth of the
subsequent wave are attached to the bone. Based on
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 EFFECT OF THYROID HORMONES 479

66 65

(а) (b)

55 54

(c) (d)

44

(e)

Fig. 3. Patterns of distribution of the pharyngeal teeth with different formulas in roach Rutilus rutilus: (a) control group, age of
308 days, 60.5 mm TL; (b) control group, age of 73 days, 27.3 mm TL; (c) TH group, age of 133 days, 33.8 mm TL; (d) TH group,
age of 390 days, 66.1 mm TL; (e) TH group, age of 285 days, 51.2 mm TL. Scale bars: (a, c–e) 200 µm; (b) 100 µm.

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480 BOLOTOVSKIY, LEVIN

II

IV

I
II

IV
III

III

Fig. 4. Initial development of the pharyngeal teeth in roach Rutilus rutilus; THIO group, age of 17 days, 6.6 mm TL: tooth I is the
first replacement wave (generation); teeth II and III are the second wave; tooth IV is the third wave (in the later development, this
tooth substitutes tooth I). Scale bar: 30 µm.

the presence of several rows of larval pharyngeal teeth
(and only one definitive row of pharyngeal teeth),
some investigators consider this character as the prim
itive state. For example, according to Vasnetsov
(1939), decrease of the number of teeth rows during
development represents the anaboly. Following Hei
ncke (1892) and Regan (1911), he proposed that the
distribution of pharyngeal teeth in several rows in
Eventognathi can be regarded as a primitive state.
However, according to Nakajima (1979, 1984, 1987),
the distribution of pharyngeal teeth in several rows
during larval development is not connected with the
distribution of these teeth in several rows in adult indi
viduals. Based on his opinion, small (minor) rows of
pharyngeal teeth represent apomorphic characters for
Cypriniformes. However, the discussion on the
derived or primitive states of the distribution of pha
ryngeal teeth (in particular, on the number of pharyn
geal teeth rows), most likely, is, far from its end (Gol
ubtsov et al., 2005; Shkil’ and Levin, 2008). We can
suggest that the distribution of the pharyngeal teeth in
roach in several rows is connected with a specific pat
tern of their development, and it is unclear if this char
acter can be regarded as a recapitulation (Fig. 4).
In the majority of experimental individuals of
roach, the teeth on the left and right pharyngeal bones
developed according to types B and D of the scheme of
Nakadjima (1984) (Fig. 5), which is apparently con
nected with directed asymmetry of the formula of pha
ryngeal teeth in this species. A similar type of develop
ment of pharyngeal teeth is observed in another repre
sentative of the subfamily Leuciscinae, bigscaled

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 EFFECT OF THYROID HORMONES 481

Po1 Ce0 An1 An2 An3 Po1 Ce0 An1 An2 An3

VIII IX iv VIII IX iv
VII iii VII iii
V VI ii V VI
IV i IV i
II III II III
I I
A B

Po1 Ce0 An1 An2 Po1 Ce0 An1 An2

VIII IX VIII IX
VII iii VII iii
V VI V VI
IV i IV
II III II III

C I D I

Fig. 5. Types (A–D) of development of the pharyngeal teeth in Cyprinidae (Nakajima, 1984). See designations in Fig. 1.

redfin Tribolodon hakonensis, with asymmetrical for
mula (54) of the main teeth row (Nakajima, 1984).
However, the suggestion on the reason of the appear
ance of directed asymmetry of the formula of pharyn
geal teeth should be tested after investigation of other
species of the subfamily Leuciscinae with similar for
mulas.
According to Nakajima (1984), types B and D lead
to the development of five and four teeth at the defin
itive state, respectively. These numbers correspond
with the numbers of teeth families during teeth devel
opment on the pharyngeal bones. In the case of for
mula 65 (as in roach), the additional teeth, most
likely, appear at later developmental stages at positions
an3 and an4. Based on the appearance of the individ
uals with definitive formula 44 in the accelerated
group, the development of the teeth on the left pha
ryngeal bone according to type C (at accelerated
development of the character) can be suggested. Fol
lowing this model of teeth development, the tooth at
position an3 is not developed, and this model is differ
ent from type B by this characteristic.
Within the largest part of the natural range, formu
las 65, and, sometimes, 55, and 66 are found in
roach (Mironovskii and Kas’yanov, 1986; Kozhara
and Kas’yanov, 1993). In the socalled “industrial”
strain of roach distributed in polluted water bodies of
Moscow, substantial differences from adjacent popu
lations by several morphological characters (including
the structure of pharyngeal teeth) are registered. In
particular, the formula 55 is observed in the “indus
trial” strain in 71% of the cases (Yakovlev, 1992). A
large frequency of occurrence of the formula 55 is
registered by Mironovskii (1994) in several roach pop
ulations of the water bodies of Moscow. As far as we
know, within the genus Rutilus, the formula 55 is typ
ical only in the roach from South Europe R. rubilio
(Müller, 1987). In this study, the action of exogenous
T3 led to the reduction of the number of pharyngeal
teeth, to increased frequency of occurrence of the for
mula 55 (up to 74.6%), and to the appearance of the
formulas 54 and 44 unusual for natural populations
of roach. However, the reasons for the deviation of the
character of the “industrial” strain of roach towards
the phenotype typical for southern species of the genus
Rutilus remain unknown. It can be suggested that
industrial pollution inducing the appearance of the
“industrial” race acts indirectly through the hypo
thalamopituitarythyroid axis.
Decrease of the numbers of teeth and teeth rows is
observed in similar hormonal experiments in African
barb L. intermedius (Smirnov and Levin, 2007; Shkil
et al., 2010). Both the reduction and increase of the
teeth and teeth row numbers in this species, most
likely, are connected with change in the rate of teeth
development on the pharyngeal bones. Change of the
ontogenetic rate induced by cultivation of the fish at
different temperatures leads to change in the number

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482 BOLOTOVSKIY, LEVIN
of pharyngeal teeth in carp Cyprinus carpio (Tatarko,
1968). However, the results were different in different
years of investigation. In the experiments with African
barb L. intermedius, development of the fish at differ
ent temperature regimes did not lead to change of the
number of pharyngeal teeth (Shkil et al., 2010). An
increase in morphological variation, including the fre
quent appearance of the formula 66 in roach, is
observed recently in the experiments on the effect of
alternating electromagnetic field on early develop
ment (Chebotareva et al., 2009). The maximum effect
by this character was obtained when the action of the
factor was applied during the interval from early gas
trulation to hatching of the embryos. We can suggest
that the general mechanism of the change of the num
ber of pharyngeal teeth is connected with heterochro
nies independently of the acting factor.
In conclusion, the development of teeth on the
pharyngeal bones in roach is accelerated after the cul
tivation of the fish in the medium with exogenous tri
iodothyronine, and it is retarded after the cultivation
of the fish in the conditions of TH deficit. The hetero
chronous development of the morphological struc
tures, most likely, is responsible for the change in the
pharyngeal teeth numbers and, consequently, for the
change of the pharyngeal teeth formulas. In addition,
the accelerated development leads to stronger change
in the teeth number in comparison to retarded devel
opment. During the larval period, the teeth of several
generations can be found on the pharyngeal bones,
and, therefore, several teeth rows appear. The pharyn
geal teeth of roach on the left and right pharyngeal
bones develop according to types B and D (Fig. 5).
However, an additional tooth develops on each (left
and right) pharyngeal bone before the reaching of the
definitive state that leads to the appearance of typical
formula 65.
ACKNOWLEDGMENTS
We thank the following colleagues for the consulta
tions and help in the study: Yu.V. Gerasimov,
Yu.G. Izyumov, M.A. Levina, E.S. Smirnova, and
S.V. Smirnov. Valuable comments on the manuscript
were made by Yu.G. Izyumov, A.V. Kozhara, and
V.N. Yakovlev. This study was conducted according to
the plan of the scientific work of the Laboratory of
Fish Ecology of the Papanin Institute for Biology of
Inland Waters (Russian Academy of Sciences) and it is
supported by a grant of the president of Russian Fed
eration for the support of young scientists (project
no. MK520.2010.4) and by the Russian Foundation
for Basic Research (project no. 080400061a).
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