Iran. J. Ichthyol. (June 2020), 7(2): 136-147 Received: February 25, 2019 © 2020 Iranian Society of Ichthyology Accepted: June 27, 2020 P-ISSN: 2383-1561; E-ISSN: 2383-0964 http://www.ijichthyol.org Research Article

Histological differences in axial musculature between relative species of the genera and (: : )

Nikita O. YABLOKOV1,2, Ivan V. ZUEV*2

1Scientific Research Institute of Ecology of Fishery Reservoirs, Krasnoyarsk, Russia. 2Siberian Federal University, Krasnoyarsk, Russia. *Email: [email protected] Abstract: Comparative studies of axial musculature in Phoxinus phoxinus and three species of the genus Rhynchocypris were conducted to analyze locomotor abilities and biotopic preferences of widespread Asian . The ratio between the areas occupied by the red and white muscles, as well as the fiber diameter, was measured on histological sections. Significant between-species differences were found in the size of the both type of fibers. Mean diameter of white fibers varied from 50.7 to 69.3μm, the extreme values (min-max) of this parameter were found in typical lotic species, R. lagowskii and P. phoxinus, respectively. Mean diameter of red fibers varied from 26.4 to 33.5μm, and also does not have a clear link to the type of habitat. On the contrary, the distribution of different types of fibres was in good agreement with the data on preferred biotopes of studied species, divided into two groups. Mean proportion of red fibres in caudal peduncle in lotic P. phoxinus and R. lagowskii (9.1- 10.2%) was more than twice as high as that in lentic R. percnurus and R. czekanowskii (3.8- 4.3%). The low proportion of red muscles in R. percnurus and R. czekanowskii, in addition to poor mobility, may also indicate adaptation to wintering under hypoxic conditions. In general, the ratio of red to white muscles provides additional information for predicting the distribution of minnows in freshwaters. Keywords: Minnows, Fibre size, Red muscles, White muscles, Preferred biotopes, Northern Asia. Citation: Yablokov, N.O. & Zuev, I.V. 2020. Histological differences in axial musculature between relative species of the genera Phoxinus and Rhynchocypris (Actinopterygii: Cypriniformes: Leuciscidae). Iranian Journal of Ichthyology 7(2): 136-147.

Introduction Such uneven distribution of different species Minnows of the genera Phoxinus and Rhynchocypris from the center of the origin cannot be fully are widespread in the Palearctic, which is inhabited explained by either theory of ocean level fluctuation by 7-9 species of Phoxinus and 5-6 species of in the quaternary period (Lindberg 1972) or Rhynchocypris (Sakai et al. 2006; Kottelat & Freyhof molecular phylogenetic data (e.g. Imoto et al. 2013; 2007; Schönhuth et al. 2018). Ranges of the Eurasian Schönhuth et al. 2018). An alternative approach to minnow, P. phoxinus and most of the Rhynchocypris the analysis of the modern ranges of species, as well species overlap in the Russian Far East which is as the forecast of their modification due climate considered to be the center of speciation of change, is to study their certain ecological niches (Yu Leuciscidae (Ito et al. 2002; Sakai et al. 2006; Perea et al. 2013). et al. 2010). The area outside the region to the west is morphology is a basic and simple tool for inhabited only by R. czekanowskii and R. lagowskii determining the ecological niches in closely related (to the East ), R. percnurus (to the East species (Gatz Jr 1981; Douglas & Matthews 1992). Europe) and the Eurasian minnow (to the Iberian However, in the case of minnows from the genus Peninsula) (Berg 1948; Kottelat & Freyhof 2007). Rhynchocypris, which are very similar in 136

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appearance, the external morphology for the given purpose is not effective. At the same time, external similarity does not exclude possible differences in the structure of the axial muscles. Axial musculature of many is distinctively divided at least into two types of muscle: red oxidative and white glycolytic. Red muscles are powered by oxidative phosphorylation, whereas white muscles are largely powered by anaerobic utilization of phosphocreatine, ATP and glycogen (Svendsen et al. 2015). Although any type of swimming is a complex combination of using of different muscles, red fibres are more active during sustained swimming, whereas white fibres are used at high swimming speed (Sanger & Stoiber 2001). Thereby, the ratio between these types of muscles correlates with capable of active and long- term movements and partly can predict the ecological requirements of fishes (Boddeke et al. 1959; Slijper 1963; Langerhans 2008). By now the anatomical Fig.1. Map of sampling sites in the Northern Asia. features of different types of muscles and their proportions are not known for all taxonomic groups 2016 in different water bodies of the Yenisei, Pyasina of fishes. Relatively many data have been published and Amur River basins (Fig. 1, Table 1). All the on marine species (Greeк-Walker & Pull 1975; specimens were killed by overdose of clove oil Mosse & Hudson 1977), while freshwater fishes are following the guidelines given by the Institutional poorly studied in this respect. Ethical Committee and were preserved in 4% neutral In this paper, we compare the ratio of red and formalin. Before sectioning, total length (TL, mm) white muscles and the size of their fibers among four was determined. related species of genera Phoxinus and Muscle sectioning: Fishes were cut into four sections Rhynchocypris from Siberia and the Russian Far with a blade (Zhang et al. 1996; Drazen et al. 2013). East. The aim of the comparison was to test a link Slice A was at the back of the base of the pectoral fin, between studied parameters and the type of preferred slice B was at the beginning of the base of the dorsal biotope and other environmental factors. fin, and slice C was at the end of the base of the anal fin (Fig. 2). Each section, except for the head, was Materials and Methods mounted under stereomicroscope Micromed MC2 Sample collection: The ratio between different types Zoom 2CR; the anterior part of each section was of axial muscles and their fibre diameter were photographed by digital camera ToupCam 5.1 estimated for the four species of the genera Phoxinus (ToupTek, China). For slices exceeding the field of and Rhynchocypris: Eurasian minnow P. phoxinus view of the microscope, a series of four photographs (Linnaeus, 1758), R. percnurus (Pallas, which included layers of epaxial and hypaxial 1814), Czekanowskii minnow R. czekanowskii muscles of the right and left parts of the slice was (Dybowski, 1869) and Amur minnow R. lagowskii taken. Red musculature was considered as triangular (Dybowski, 1869). Fishes were caught by patches of dark color, located laterally between the electrofishing during the summer months of 2013- layers of epaxial and hypaxial white muscle tissue 137 Iran. J. Ichthyol. (June 2020), 7(2): 136-147

Table 1. Sample sites and samples description.

River basin Geographical Type of Sampling № Species TL ± SE, mm N coordinates water body period 57°27'08.5" N, August 73.6 ± 4.2 1 Yenisei Lake 5 Rhynchocypris 93°11'03.6" E 2016 62.0 –85.0 czekanowskii 69°22'48.1"N July 90.0 ± 5.4 2 Pyasina Lake 5 88°11'41.2"E 2013 71.0 – 108.0 Rhynchocypris 56°34'57.3"N, August 60.9 ± 0.7 3 Yenisei Lake 10 percnurus 93°43'23.6"E 2016 56.0 – 64.0 Rhynchocypris 57°27'08.5"N, June 106.5± 0.4 4 Amur Reservoir 10 lagowskii 93°11'03.6"E 2014 84.0 –122.0 Phoxinus 57°27'08.5"N, July 75.9 ± 2.1 5 Yenisei River 10 phoxinus 93°11'03.6"E 2015 63.0 –87.0

Table 2. Duration of exposure of minnow’s muscle samples at the Microm STP 120 automated station.

№ Reagent Exposure, min 1 Ethanol 70° 30 2 Ethanol 80° 30 3 Ethanol 90° 30 4 Ethanol 96° 35 5 Ethanol 96° 35 6 Ethanol 100° 35 7 Ethanol 100° 35 8 Xylol 45 9 Xylol 45 10 Paraffin 60 11 Paraffin 80

Microm STP 120 (Table 2). Paraffin was poured into samples using a filling system Thermo Scientific Fig.2. Position of slices and distribution of white (1) Microm EC 350-1; sections were prepared using a and red (2) muscles. semi-automatic microtome Microm HM 440E. (Fig. 2). The area occupied by the red and white Thickness of histological sections was 10μm. muscles was measured in photographs in the ImageJ Obtained histological sections were viewed under 1.51r graphics editor (NIH, USA). Proportion of red a Leica DMLC light microscope equipped with a muscles was estimated as a percentage of the total digital camera (Leica DC100). Photographing of area occupied by the muscles in the cross section for sections was performed under different each slice. magnification. The shortest fiber diameter, calculated Histological preparation. About 1cm3 size blocks perpendicular to the longest diameter at its mid-point, were extracted from section B-C (Fig. 2). For the was measured by digital image using ImageJ 1.51r histology examination, samples were dehydrated, software (NIH, USA). For each examined fish, the cleared with xylol, then embedded in paraffin and diameters of 50 red and 50 white fibres were stained with hematoxylin and Ehrlich’s eosin measured at random. (Mumford 2004). Dehydration was carried out by Statistical analyses. Statistical analyses were incubation in ascending concentrations of alcohols performed with PAST 3.17 (Hammer et al. 2001). (xylol and ethanol) using an automatic station Fibre diameter and red muscle proportion is

138 Yablokov and Zuev - Histological differences in axial musculature

Table 3. Red and white fibre diameter of four species of minnows.

Fibre diameter (mean ± SE / range, in μm) Fibre F-value P-value type Rhynchocypris Rhynchocypris Phoxinus Rhynchocypris czekanowskii percnurus phoxinus lagowskii 60.4 ± 1.8B 63.9 ± 2.1BC 69.3 ± 2.8C 50.7 ± 1.6A White 11.4 0.00 25–130 20–123 16–147 23 – 85 31.8 ± 0.6B 26.4 ± 2.1A 33.5 ± 0.7B 32.3 ± 0.8B Red 17.3 0.00 14 – 48 11 – 45 17 – 55 15 – 65

Table 4. Proportions of red muscles in different parts of axial muscles of four species of minnows.

Proportions of red muscles (mean ± SE / range, in %) Slice Rhynchocypris czekanowskii Rhynchocypris Phoxinus Rhynchocypris F-value P-value 1 2 percnurus phoxinus lagowskii 2.0±0.2A 2.5±0.3A 3.2±0.2AB 4.8±0.4C 4.3±0.3BC A 13.4 0.00 1.6–2.7 1.9–3.4 2.0–4.0 2.9–7.1 3.2–5.6 2.3±0.3A 2.6±0.1A 2.5±0.2A 4.1±0.2B 4.3±0.3B B 12.4 0.00 1.5–3.3 2.3–3.0 1.5–3.7 2.5–5.2 2.9–5.8 3.8±0.3A 4.1±0.4A 4.3±0.4A 9.1±0.6B 10.2±0.7B C 32.0 0.00 3.2–4.2 2.9–5.0 2.5–5.8 6.1–12.9 7.3–14.4 expressed as mean±standard error (SE). ANOVA was conducted to detect interspecies differences in the measured parameters. When F-test indicated significance, species means were compared by a Tukey’s test.

Results The white muscles of the studied minnow species are represented by angular fibres with irregular shape (Fig. 3). White fibres occupy most of the area of the axial muscles in all sections of the minnows, ranging from 90 to 98% of the total musculature in the cross Fig.3. Histological sections of the muscular tissue of section of the body. The red muscles are represented Phoxinus phoxinus: A, white fibres; B, red fibres. by fibres having a rounded shape on histological sections (Fig. 3). This type of muscle fibres is located narrowest range of fibre diameters (from 23 to laterally, between the epaxial and hypaxial parts of 85μm), while other minnow species had fibres larger myomere. Mean diameter of the white fibres was 60- than 100μm (Table 3). 69µm with a range of 16-147µm. The diameter of the Mean diameter of the red fibres was 1.6-2.4 times white fibres varied great for all species. P. phoxinus smaller than the diameter of the white fibres and was had the highest mean diameter of white fibres equal to 26-33μm. The range of diameters of red (69.3μm), while R. lagowskii had the smallest value fibres was also significantly lower than that of white of this parameter (50.7 μm). For R. percnurus and R. fibres and was represented by fibres from 11 to czekanowskii, the mean white fibre diameters were 65μm. The diameter of the red fibres varied slightly. 63.9 and 63.4μm, respectively. It should also be There were no differences in the parameter between noted that R. lagowskii was characterized by the species, except R. percnurus, characterized by the 139 Iran. J. Ichthyol. (June 2020), 7(2): 136-147

Table 5. Mean proportion of red muscles in caudal peduncle of some species from Leuciscidae subfamily. Species Mean, in % Reference chiliticus (Cope, 1870) 12.0 Gatz 1979 ardens (Cope, 1868) 11.1 Gatz 1979 leuciscus (Linnaeus, 1758) 10.8 Yablokov & Zhukova 2017 rutilus (Linnaeus, 1758) 10.3 Broughton et al. 1981 Rhynchocypris lagowskii (Dybowski, 1869) 10.2 Our data analostana Girard, 1859 9.9 Gatz 1979 atromaculatus (Mitchill, 1818) 9.2 Gatz 1979 Phoxinus phoxinus (Linnaeus, 1758) 9.1 Our data cataractae (Valenciennes, 1842) 9.0 Gill et al. 1989 hypsinotus (Cope, 1870) 6.7 Gatz 1979 (Phoxinus) oreas Cope, 1867 6.1 Gatz 1979 Notropis alborus Hubbs & Raney, 1947 5.8 Gatz 1979 robusta Baird & Girard, 1853 5.9 Moran et al. 2016 Gila elegans Baird & Girard, 1853 5.3 Moran et al. 2016 Rhynchocypris percnurus (Pallas, 1814) 4.3 Our data Rhynchocypris czekanowskii (Dybowski, 1869) 3.8-4.1 Our data notatus (Rafinesque, 1820) 3.8 Gill et al. 1982 smallest value of this parameter (Table 3). related species of minnows were observed both in The proportion of red muscles varied for different fiber size and in the ratio of red and white muscles. parts of the body. For all species, maximal proportion However, differences in one group of traits did not of red fibres was in the caudal peduncle (slice C) correspond to others. According to the ratio of red which exceeded this parameter in others slices by and white fibers, the studied species were clearly 1.5-2 times (Fig. 2, Table 4). Proportions of red divided into two groups: R. percnurus and muscles on slices A and B in the R. lagowskii and R. czekanowskii versus R. lagowskii and R. czekanowskii did not differ; in the P. phoxinus and P. phoxinus. The Amur and Eurasian minnows were R. percnurus, the proportion of red muscles in slice distinguished by the minimum and maximum mean A was 1.0% higher than in slice B. The studied diameter of white fibers; lake minnow was species can be divided into 2 groups according to the distinguished by the minimum mean diameter of red proportion of red muscles. One group consists of lake fibers. and Czekanowskii’s minnows, in which the The growth of both types of muscles is determined proportion of the muscles in the trunk region (slices by the combination of enlargement of the muscle A and B) varied between 2.0-3.2%, and about 4% in fibres already existing (hypertrophy) and the the caudal peduncle (Table 4). Eurasian and Amur recruitment of new fibres (hyperplasia) (Stoiber et al. minnows form the other group, having red muscle 2002). Hyperplasia usually take place in the early proportion in the trunk region about 4%, and in stages of development, while hypertrophic growth caudal peduncle – about 9-10%. can occur at all stages of fish development (Stickland 1983; Rowlerson et al. 1995; Zimmerman & Lowery Discussion 1999). This makes it difficult to compare fiber In the present study, histological differences between diameter in fish of different sizes or ages. The

140 Yablokov and Zuev - Histological differences in axial musculature

species-specific ratio of red and white muscles, on preferred biotopes. The Eurasian minnow lives in a the other hand, is established early and remains stable wide range of cold and well oxygenated habitats, with age (Higgins & Thorpe 1990; Cediel et al. from small, fast-flowing streams to large lowland 2008). rivers (Frost 1943; Kottelat & Freyhof 2007; Kirillov Despite the revealed differences in fiber sizes in & Knizhin 2014). It also can inhabit flowing minnows, their values are in the range for most fish. oligotrophic lakes and brackish coastal waters (Zuev Generally, diameters of white muscle fibres of fishes et al. 2012; Museth et al. 2007; Svirgsden et al. 2016). are on average about 70µm with the range of 20-300 The species is characterized by spawning and feeding µm (Bone 1978; Sanger & Stoiber 2001). Red muscle migrations, even between sea brackish water and fibres vary to a lesser extent (25-45µm on average), rivers (Frost 1940; Svirgsden et al. 2016). Holthe et possibly because there are physical limits on the size al. (2009) showed that large individuals of the of the cell above which diffusion processes are too minnow could maintain a high swimming speed of slow to permit continuous aerobic activity (Greer- 34cm s-1 for at least 25 min. Walker & Pull 1975). The Amur minnow usually inhabits biotopes It is considering that there is no regular similar to the Eurasian minnow (Nikitin & Safronov relationship between the level of swimming activity 2009; Antonov 2012; Kirillov & Knizhin 2014). In in fish and the size of muscle fibres (Greer-Walker & areas of overlapping ranges of two species they can Pull 1975). However, in an experiment with gilthead form mixed schools (Sverdlova & Knizhin 2011). sea bream was shown that sustained exercise The species can also coexist with the Chinese or enhances hypertrophic development of white muscle upstream fat minnow R. oxycephalus, which typical (Ibarz et al. 2011). It is possible, that the large size of habitats are usually restricted in cold headwaters (Yu white fibers in the mobile Eurasian minnow reflects et al. 2013; Nishida et al. 2014, 2015). Nishida et al. this pattern. However, lotic species - Amur minnow, (2015) shown, that R. oxycephalus can replace had the smallest fiber size. R. lagowskii in the upstream section of a river with In contrast to the previous one, a large number of lower temperatures. studies demonstrate a clear link between the In contrast to the previous species, the lake proportion of red muscles and various expressions of minnow lives mainly in small, shallow and often swimming activity, in terms of riverine vs. lacustrine, isolated water bodies (Kottelat & Freyhof 2007; benthic vs. pelagic, sedentary vs. mobile, migratory Kaminski et al. 2011; Kusznierz et al. 2011; Kirillov vs. non-migratory (Greer-Walker & Pull 1975; & Knizhin 2014). These biotopes are characterized Meyer-Rochow & Ingram 1993; Drazen et al. 2013). periodically by low oxygen saturation and/or low pH For the representatives of the Leuciscidae, data on values (Kamiñski et al. 2011). axial muscles distribution is known primarily for The Czekanowskii’s minnow is the least studied North American species. Among them the mean species among Asian minnows. The species can percentage of red muscle in caudal peduncle varies inhibit wide range of water bodies, although three times, from 4 to 12% (Table 5). Minnows with information on preferred biotopes is inconsistent. 9-12% of the parameter inhabit mainly flowing There is an opinion that life history traits of the waters like creeks and rivers (Froese & Pauly 2018). Czekanowskii’s minnow are close to those of the Other species, with a share of red muscles from 3 to Eurasian minnow (Atlas of Freshwater Fish of Russia 6%, are considered to be equally riverine and 2002). But in the Yenisei and Pyasina river basins we lacustrine (Froese & Pauly 2018). have never found the species in fast flowing rivers The position of the studied minnows at different with stony ground, where P. phoxinus lives. parts of the table corresponds to different types of According to our research, R. czekanowskii 141 Iran. J. Ichthyol. (June 2020), 7(2): 136-147

inhabits mainly lentic biotopes, like small lakes and (2016) on minnows in shallow, ice-covered Canada slow flowing small rivers (Zuev et al. 2006, 2019). streams show that during winter fishes migrate to On the whole, as it is based on our results and the deep pools, with low-velocities, but remain active. literature data, minnows of the genus Rhynchocypris Among them, Semotilus atromaculatus was the most can be virtually distributed across flow regimes as common species, which has 9.2% red muscle in follows: R. percnurus, R. czekanowskii, R. lagowskii caudal peduncle. The Eurasian minnow also and R. oxycephalus. This sequence does not maintains swimming activity during winter months correspond to the phylogeny of species. According to (Frost 1940). In the study of Radtke (2011) the lake Sakai et al. (2006), R. oxycephalus is closer to minnow from northern was active during the R. percnurus and R. czekanowskii than to entire ice-free period and even under full ice cover in R. lagowskii. March. However, in Northern Asia ice-cover period Despite the obvious divergence of the minnows as lasts longer, usually from October to April. Malyshev to preferred biotopes, each of the species examined (1982) reports on the findings of the Lake Minnow may inhabit both rivers and lakes. Presumably, the individuals in state of anabiosis in a small Siberian link between red muscle proportion and the types of lake in March. We also found inactive individuals of biotopes occurs most contrastingly at low water R. czekanowskii buried in the lake sediments in the temperatures. As it is indicated by Maxent model, early November (Zuev, unpubl. data). minimum temperature of the coldest month most Thus, a combination of several environmental seriously influenced the distribution of factors (flow regime, water temperature, dissolved R. oxycephalus (Yu et al. 2013). All investigated oxygen concentration) can affect certain ratio minnows also live in waters with significant annual between red and white axial muscles in minnows. temperature range (exceeding 15°C), that requires There is trade-off not only between living in biotopes compensatory responses to thermal acclimation with different flow regimes, but also between achieved in many ways. One of the ways found in wintering in biotopes with different dissolved oxygen is increasing the proportion of red levels. As shown Cech et al. (1990), who had studied muscles (Guderley & Blier 1988). Higher proportion distribution of fishes in California streams along its of red muscle should support locomotory capacity in metabolic rates, differential metabolic responses to flowing water at low temperatures (Guderley & Blier hypoxia increase the complexity of pattern of 1988; Cech et al. 1990). distribution. However, in shallow lakes, which are typical In conclusion, we have identified differences in biotopes for R. percnurus and R. czekanowskii, low the structure of the axial muscles, which can temperatures are usually combined with hypoxia or determine the ecological niche of closely related and anoxia conditions. A key adaptation for survival with similar in external morphology species of genera oxygen deficiency is strong metabolic depression (to Phoxinus and Rhynchocypris. Among the studied approximately 30% of normal), as it was shown for parameters, the greatest differences were found in the Carassius species (Bickler & Buck 2007). Even ratio of red and white muscles in the caudal peduncle under hypoxic conditions, some species can maintain of fish. Mean proportion of red fibres in caudal activity (e.g. Carassius auratus; Nilsson 2001); others peduncle in lotic P. phoxinus and R. lagowskii (9.1- become dormant in cold months (Crawshaw 1980). 10.2%) was more than twice as high as that in lentic In the dormancy state, macromolecular reserve R. percnurus and R. czekanowskii (3.8-4.3%). becomes the main role of the muscles. Despite the obvious connection between the ratio of Information on the behavior of the minnows red and white muscles of minnows with the type of during winter is insufficient. Davis observations biotope, we cannot estimate the relative contribution 142 Yablokov and Zuev - Histological differences in axial musculature

of additional factors (water temperature and fish. American Journal of Physiology-Regulatory, dissolved oxygen concentration) to the development Integrative and Comparative Physiology 246(4): 479- of different types of musculature. For clarification, 486. direct observations of the way of life in the ice- Davis, L. 2016. Overwinter habitat of minnows in small, covered period and the resistance to hypoxia of each southern Ontario streams. Guelph, Ontario, Canada. Douglas, M.E. & Matthews, W.J. 1992. Does of the species are required. morphology predict ecology? Hypothesis testing within a freshwater stream fish assemblage. Oikos Acknowledgements 65(2): 213-224. The work was supported by the project No. Drazen, J.C.; Dugan, B. & Friedman, J.R. 2013. Red 6.1504.2017/4.6 of Siberian Federal University muscle proportions and enzyme activities in deep‐sea carried out according to Federal Tasks of Ministry of demersal fishes. Journal of Fish Biology 83(6): 1592- Education and Science of Russian Federation. 1612. Froese, R. & Pauly, D. 2019. Fishbase. World Wide References WebElectronic Publication. www.fishbase.org Antonov, A.L. 2012. Diversity of fishes and structure of (accessed 15 January 2019). ichthyocenoses in mountain catchment areas of the Frost, W.E. 1943. The natural history of the minnow, Amur Basin. Journal of ichthyology 52 (2): 149-159. Phoxinus phoxinus. The Journal of Ecology Atlas of Freshwater Fish of Russia. 2002. Nauka publ, 12(2): 139-162. Moscow, Vol.1. (In Russian) Gatz Jr, A.J. 1981. Morphologically inferred niche Berg, L.S. 1940. Classification of fishes, both recent and differentiation in stream fishes. American Midland fossil. USSR Academy of Sciences Publishing, Naturalist 106(1): 10-21. Moscow. (In Russian) Gatz, J.A. 1979. Ecological morphology of freshwater Bickler, P.E. & Buck, L.T. 2007. Hypoxia tolerance in stream fishes. Tulane studies in zoology and botany reptiles, amphibians, and fishes: life with variable 21(2): 91-124. oxygen availability. Annual Review of Physiology Gill, H.S.; Weatherley, A.H. & Bhesania, T. 1982. 69: 145-170. Histochemical characterization of myotomal muscle Boddeke, R.; Slijper, E.J. & Van der Stelt, A. 1959. in the bluntnose minnow, Pimephales notatus Histological characteristics of the body musculature Rafinesque. Journal of Fish Biology 21(2): 205-214. of fishes in connection with their mode of life. Gill, H.S.; Weatherley, A.H.; Lee, R. & Legere, D. 1989. Koninklijke Nederlandse Akademie van Histochemical characterization of myotomal muscle Wetenschappen. Proceedings. Ser. C. 62: 576-588. of five teleost species. Journal of fish biology 34(3): Broughton, N.M.; Goldspink, G. & Jones, N.V. 1981. 375-386. Histological differences in the lateral musculature of Greer‐Walker, M. & Pull, G.A. 1975. A survey of red O‐group roach, Rutilus rutilus (L.) from different and white muscle in marine fish. Journal of Fish habitats. Journal of Fish Biology 18(2): 117-122. Biology 7(3): 295-300. Cech, J.J.; Mitchell, S.J.; Castleberry, D.T. & McEnroe, Guderley, H. & Blier, P. 1988. Thermal acclimation in M. 1990. Distribution of California stream fishes: fish: conservative and labile properties of swimming influence of environmental temperature and hypoxia. muscle. Canadian Journal of Zoology 66(5): 1105- Environmental biology of Fishes 29(2): 95-105. 1115. Cediel, R.A.; Blob, R.W.; Schrank, G.D.; Plourde, R.C. Hammer, Ø.; Harper, D.A.T. & Ryan, P.D. 2001. PAST: & Schoenfuss, H.L. 2008. Muscle fiber type Paleontological statistics software package for distribution in climbing Hawaiian gobioid fishes: education and data analysis. Palaeontologia ontogeny and correlations with locomotor Electronica 4(1): 9 p. performance. Zoology 111(2): 114-122. Higgins, P.J. & Thorpe, J.E. 1990. Hyperplasia and Crawshaw, L.I. 1984. Low-temperature dormancy in hypertrophy in the growth of skeletal muscle in

143 Iran. J. Ichthyol. (June 2020), 7(2): 136-147

juvenile Atlantic salmon, Salmo salar L. Journal of forest zone of Western Siberia. In: Krivoshchekov, Fish Biology 37(4): 505-519. G.M. (ed.), A complex study and exploitation of the Holthe, E.; Lund, E.; Finstad, B.; Thorstad, E.B. & Karasuk Lakes. Nauka publ., Siberian branch, McKinley, R.S. 2009. Swimming performance of the Novosibirsk. pp. 173-203. (In Russian) European minnow. Boreal Environment Research Meyer-Rochow, V.B. & Ingram, J.R. 1993. Red-white 14(2): 272-278. muscle distribution and fibre growth dynamics: a Ibarz, A.; Felip, O.; Fernández-Borràs, J.; Martín-Pérez, comparison between lacustrine and riverine M.; Blasco, J. & Torrella, J.R. 2011. Sustained populations of the Southern Retropinna swimming improves muscle growth and cellularity in retropinna Richardson. Proceedings of the Royal gilthead sea bream. Journal of Comparative Society of London 252(1334): 85-92. Physiology B 181(2): 209-217. Moran, C.J.; Ferry, L.A. & Gibb, A.C. 2016. Why does Imoto, J.M.; Saitoh, K.; Sasaki, T.; Yonezawa, T.; Gila elegans have a bony tail? A study of swimming Adachi, J.; Kartavtsev, Y.P.; Miya, M.; Nishida, M. morphology convergence. Zoology 119(3): 175-181. & Hanzawa, N. 2013. Phylogeny and biogeography Mosse, P.R.L. & Hudson, R.C.L. 1977. The functional of highly diverged freshwater fish species roles of different muscle fibre types identified in the (, Cyprinidae, Teleostei) inferred from myotomes of marine teleosts: a behavioural, mitochondrial genome analysis. Gene 514(2): 112- anatomical and histochemical study. Journal of Fish 24. Biology 11(5): 417-430. Ito, Y.; Sakai, H.; Shedko, S. & Jeon S.R. 2002. Genetic Mumford, S.L. 2009. Histology of finfish. In: Heil, N. differentiation of the northern Far East cyprinids, (ed.), National Wild Fish Health Survey – Laboratory Phoxinus and Rhynchocypris. Fisheries Science 68 Procedures Manual, 5.0 Edition. U.S. Fish and (1): 75-78. Wildlife Service. Warm Springs, GA, USA. pp. 367- Kaminski, R.; Wolnicki, J. & Sikorska, J. 2011. Physical 380. and chemical water properties in water bodies Museth, J.; Hesthagen, T.; Sandlund, O.T.; Thorstad, E. inhabited by the endangered lake minnow, B. & Ugedal, O. 2007. The history of the minnow Eupallasella percnurus (Pall.), in central Poland. Phoxinus phoxinus (L.) in Norway: from harmless Archives of Polish Fisheries 19(3): 153-159. species to pest. Journal of Fish Biology 71: 184-195. Kirillov, A.F. & Knizhin, I.B. 2014. Ichthyofauna of the Nikitin, V.D. & Safronov, S.N. 2009. History of studies, Lena River (Laptev Sea Basin): modern composition species composition, morphology and distribution of and historical formation. Journal of Ichthyology minnows from the genius Rhynchocypris 54(7): 433-445. (Cyprinidae) on Sakhalin island. The Bulletin of Kottelat, M. & Freyhof, J. 2007. Handbook of European Irkutsk State University. Series Biology. Ecology freshwater fishes. Publications Kottelat, Cornol and 2(2): 41-44. (In Russian) Freyhof, Berlin. Nilsson, G.E. 2001. Surviving anoxia with the brain Kusznierz, J.; Paśko, Ł. & Tagayev, D. 2011. On the turned on. News in Physiological Sciences 16(5): variation and distribution of the lake minnow, 217-221. Eupallasella percnurus (Pall.). Archives of Polish Nishida, K.; Koizumi, N.; Satoh, T.; Senga, Y.; Fisheries 19(3): 161-166. Takemura, T.; Watabe, K. & Mori, A. 2015. Influence Langerhans, R.B. 2008. Predictability of phenotypic of the domestic alien fish Rhynchocypris differentiation across flow regimes in fishes. oxycephalus invasion on the distribution of the Integrative and Comparative Biology 48(6): 750-768. closely related native fish R. lagowskii in the Tama Lindberg, G.U. 1972. Large Fluctuations in the Ocean River Basin, . Landscape and Ecological Level in the Quaternary Period. Nauka, Leningrad. Engineering 11(1): 169-176. (In Russian) Nishida, K.; Ohira, M. & Senga, Y. 2014. Movement and Malyshev, Y.F. 1982. Ecology of lake minnow Phoxinus assemblage of fish in an artificial wetland and canal percnurus (Pallas) of water reservoirs in the steppe- in a paddy fields area, in eastern Japan. Landscape

144 Yablokov and Zuev - Histological differences in axial musculature

and Ecological Engineering 10(2): 309-321. different temperatures: simulating natural Perea, S.; Böhme, M.; Zupančič, P.; Freyhof, J.; Šanda, environmental conditions for a temperate freshwater R.; Özuluğ, M.; Abdoli, A. & Doadrio, I. 2010. cyprinid. Journal of Experimental Biology 205(16): Phylogenetic relationships and biogeographical 2349-2364. patterns in Circum-Mediterranean subfamily Svendsen, J.C.; Tirsgaard, B.; Cordero, G.A. & Leuciscinae (Teleostei, Cyprinidae) inferred from Steffensen, J.F. 2015. Intraspecific variation in both mitochondrial and nuclear data. BMC aerobic and anaerobic locomotion: gilthead sea bream Evolutionary Biology 10(265): 1-27. (Sparus aurata) and Trinidadian guppy (Poecilia Radtke, G. 2011. Comparison of seasonal activity of the reticulata) do not exhibit a trade-off between lake minnow, Eupallasella percnurus (Pall.), and maximum sustained swimming speed and minimum crucian , Carassius carassius (L.), in small water cost of transport. Frontiers in Physiology 6(43): 1-12. bodies in northern Poland. Archives of Polish Sverdlova, T.V. & Knizhin, I.B. 2011. The biology of Fisheries 19(3): 175-182. three species minnows of the upper stream of the Rowlerson, A.; Mascarello, F.; Radaelli, G. & Veggetti Lena river. Baikal Zoological Journal 3: 42-48. (In A. 1995. Differentiation and growth of muscle in the Russian) fish Sparus aurata (L): II. Hyperplastic and Svirgsden, R.; Rohtla, M.; Albert, A.; Taal, I.; Saks, L.; hypertrophic growth of lateral muscle from hatching Verliin, A. & Vetemaa, M. 2018. Do Eurasian to adult. Journal of Muscle Research and Cell minnows (Phoxinus phoxinus L.) inhabiting brackish Motility 16(3): 223-236. water enter fresh water to reproduce: Evidence from Sakai, H.; Ito, Y.; Shedko, S.V.; Safronov, S.N.; Frolov, a study on otolith microchemistry. Ecology of S.V.; Chereshnev, I.A.; Jeon, S. & Goto, A. 2006. Freshwater Fish 27(1): 89-97. Phylogenetic and taxonomic relationships of northern Yablokov, N.O. & Zhukova, K.A. 2017. Morphological Far Eastern phoxinin minnows, Phoxinus and features of red and white musculature of some Rhynchocypris (Pisces, Cyprinidae), as inferred from representatives of ichthyofauna from the Middle allozyme and mitochondrial 16S rRNA sequence Yenisei. In: Modern problems and prospects for the analyses. Zoological Science 23(4): 323-331. development of the fisheries complex: materials of Sanger, A.M. & Stoiber, W. 2001. Muscle fiber diversity the Vth Scientific and Practical Conference of Young and plasticity. In: Johnston, I.A. (ed.), Fish Scientists with International Participation, Moskow, Physiology: Muscle Development and Growth. Russia, 17-18 April 2017, pp. 305-309. (In Russian) Academic Press, San Diego, San Francisco, New Yu, D.; Chen, M.; Zhou, Z.; Eric, R.; Tang, Q. & Liu, H. York, Boston, London, Sydney, Tokyo. pp. 187-250. 2013. Global climate change will severely decrease Schönhuth, S.; Vukic, J.; Sanda, R.; Yang, L. & Mayden, potential distribution of the East Asian coldwater fish R.L. 2018. Phylogenetic relationships and Rhynchocypris oxycephalus (Actinopterygii, classification of the Holarctic family Leuciscidae Cyprinidae). Hydrobiologia 700(1): 23-32. (Cypriniformes: Cyprinoidei). Molecular Zhang, G.; Swank, D.M. & Rome, L.C. 1996. Phylogenetics and Evolution 127: 781-799. Quantitative distribution of muscle fiber types in the Slijper, E.J. 1963. Functional analysis of contractive scup Stenotomus chrysops. Journal of Morphology tissues. In: Proceedings of the 16th International 229(1): 71-81. Congress of Zoology 3, Washington, DC, USA, 20-27 Zimmerman, A.M. & Lowery, M.S. 1999. Hyperplastic August 1963, pp. 257-262. development and hypertrophic growth of muscle Stickland, N.C. 1983. Growth and development of fibers in the white seabass (Atractoscion nobilis). muscle fibres in the rainbow trout (Salmo gairdneri). Journal of Experimental Zoology 284(3): 299-308. Journal of anatomy 137(2): 323-333. Zuev, I.V.; Chuprov, S.M. & Zueva, A.V. 2019. Stoiber, W.; Haslett, J.R.; Wenk, R.; Steinbacher, P.; Expanding the geographical distribution of Gollmann, H.P. & Sanger, A.M. 2002. Cellularity Rhynchocypris czekanowskii (Dybowski, 1869) changes in developing red and white fish muscle at (Cypriniformes, Cyprinidae) in the basin of the

145 Iran. J. Ichthyol. (June 2020), 7(2): 136-147

Yenisei river, Eastern Siberia, Russia. Check List 15(3): 369-374. Zuev, I.V.; Dubovskaya, O.P.; Ivanova, E.A.; Gluschenko, L.A.; Shulepina, S.P. & Ageev, A.V. 2012. Evaluation of the potential fish productivity of lake Oiskoe (Ergaky Mountain Range, West Sayan) basing on food supply. Contemporary Problems of Ecology 5(4): 470-479. Zuev, I.V.; Vyshegorodtsev, A.A. & Diterle, A.V. 2006. The morphological and ecological characterization of Chekanovsky minnow Phoxinus czekanowskii, Dybowski (Cyprinidae: Cypriniformes) in water bodies of the basin of the Enisey and Pyasina Rivers (East Siberia). Contemporary Problems of Ecology 4: 511-520. (In Russian)

146 Iran. J. Ichthyol. (June 2020), 7(2): 136–147 Received: February 25, 2019 © 2020 Iranian Society of Ichthyology Accepted: June 27, 2020 P-ISSN: 2383-1561; E-ISSN: 2383-0964 http://www.ijichthyol.org

مقاله پژوهشی تفاوتهای بافتشناسی در عضله محوری بین گونههای خویشاوند جنسهای Phoxinus و Rhynchocypris )شعاع بالگان: کپورماهی شکالن: لوسیسیده(

نیکیتا او. یابلوکو1،2، ایوان وی. زوئف2* 1پژوهشکده علمی پژوهشی بوم شناسی ذخایر شیالتی، کراسنویارسک، روسیه. 2دانشگاه فدرال سیبری، کراسنویارسک، روسیه.

چکیده: مطالعات مقایسهای عضله محوری در Phoxinus phoxinus و سه گونه از جنس Rhynchocypris برای تجزیه و تحلیل قابلیتهای حرکتی و ترجیحات بیوتوپیک آنها انجام شد. نسبت بین نواحی اشغال شده توسط ماهیچههای قرمز و سفید و همچنین قطر فیبرها در برشهای بافتی اندازهگیری شد. اختالفات معنیداری در اندازه هر دو نوع فیبر در بین گونهها مشاهده شد. متوسط قطر فیبرهای سفید از 7/50 تا 3/69 میکرومتر متغیر بود، و بیشینه و کمینه این پارامتر به ترتیب در گونههای لوتیک R. lagowskii و P. phoxinus یافت شد. قطر متوسط فیبرهای قرمز از 4/26 تا 5/33 میکرومتر متغیر است، و همچنین ارتباط مستقیمی با نوع زیستگاه ندارد. در مقابل، توزیع انواع مختلف فیبر مطابق با دادههای موجود در مورد بیوتوپهای مورد ترجیح گونههای مورد مطالعه مطابقت خوبی داشت و به دو گروه تقسیم میشدند. میانگین نسبت فیبرهای قرمز در ساقه دمی در گونه های لوتیک P. phoxinus و R. lagowskii )2/1-10/9 درصد( بیش از دو برابر آن در گونههای لنتیک R. percnurus و R. czekanowskii )3/8-4/3 درصد( بود. نسبت پایین عضالت قرمز در R. percnurus و R. czekanowskii عالوه بر تحرك ضعیف، ممکن است نشان دهنده سازگاری با زمستان در شرایط هیپوکسی نیز باشد. به طور کلی، نسبت ماهیچه های قرمز به سفید اطالعاتی تکمیلی جهت پیش بینی توزیع مینوها در آبهای شیرین را فراهم میسازد. کلماتکلیدی: کپورها، اندازه فیبر، عضالت قرمز، عضالت سفید، بیوتوپهای ترجیحی، آسیای شمالی.

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