Boule gene expression underpins the meiotic arrest in spermatogenesis in male rainbow trout (Oncorhynchus mykiss) exposed to DEHP and butachlor.

Boule, the ancestor of the DAZ (Deleted in AZoospermia) gene family, in most organisms is mainly involved in male meiosis. The present study investigates the effects of the plasticizer DEHP (50mg/kg body weight) and herbicide butachlor (0.39mg/L) on male rainbow trout (Oncorhynchus mykiss) for a 10-day period in two independent experiments. The results showed that plasma testosterone (T) concentrations were significantly lower in fish exposed to either DEHP or butachlor compared to the control fish (P<0.05). Fish showed a significantly elevated hepatosomatic index (HSI) in the butachlor treatment (P<0.05). However, no significant difference was observed in HSI values in the DEHP treatment (P>0.05). In addition, no significant differences were found in the gonadosomatic index (GSI) in both DEHP and butachlor treatments (P>0.05). Histologically, testes of male trout in the control groups were well differentiated and filled with large numbers of cystic structures containing spermatozoa. In contrast, the testes of male trout contained mostly spermatocytes with few spermatozoa in both treated group, suggesting that DEHP and butachlor may inhibit the progression of meiosis. Also, boule gene expression was significantly lower in the testes of male trout affected by DEHP and butachlor in comparison with their control groups (P<0.05), which confirmed the meiotic arrest in affected trout. Based on the results, the present study demonstrated that DEHP and butachlor can inhibit the progression of spermatogenesis in male trout, potentially by causing an arrest of meiosis, maybe due to down-regulation of boule gene expression through T and/or IGF1 via ERK1/2 signaling in T-independent pathways. In addition, these results confirmed that boule can be considered as a predictive marker to assess meiotic efficiency.

Using the whole body as a sucker: combining respiration and feeding with an attached lifestyle in hill stream loaches (Balitoridae, Cypriniformes).

Small fishes living in fast-flowing rivers face a harsh environment as they can easily be swept away by the rapid currents. To survive such circumstances, teleosts evolved a wide variety of attachment mechanisms, based on friction, negative pressure or both. Balitorinae (Balitoridae, Cypriniformes) are exceptional in using their whole body as an adhesive apparatus. We investigated the morphological adaptations of Balitorinae by studying the osteology and myology of four species (Beaufortia leveretti, Sewellia lineolata, Pseudogastromyzon myersi, and Gastromyzon punctulatus) using clearing and staining, serial cross-sections and CT-scanning. A kinematic analysis was performed to study the respiration and feeding mechanisms and to identify key structures in these mechanisms. Our research showed that the whole body of Balitorinae acts as a suction disc, with friction-enhancing structures (unculi) on the thickened anterior rays of the paired fins. The abruptly rising head profile, supported by the extremely enlarged lacrimal bone and the flat ventral body surface facilitate effective substrate attachment. During attachment, the pelvic girdle is pulled anterodorsally, suggesting the formation of a negative pressure underneath the body. Detachment by water inflow underneath the body is prevented by three mechanisms. 1) Barbels control the water inflow by detachment and reattachment to the substrate. 2) Most water present underneath the body is removed during inspiration. 3) Excess water is regularly removed by movements of the posterior pectoral fin rays. The balitorine body is thus modified as such that it allows effective attachment, while not impairing respiration. Comparison with other teleosts living in similar environments shows that most species use more locally concentrated modifications of the paired fins and/or the mouth for attachment. The high diversity in teleostean adhesive apparatuses and associated myological modifications suggest a substantial functional convergent evolution, without necessarily highly convergent anatomical adaptations.

Functional and evolutionary anatomy of the African suckermouth catfishes (Siluriformes: Mochokidae): convergent evolution in Afrotropical and Neotropical faunas.

Of those fishes scraping food off substrates and using head parts in substrate attachment for station-holding, the catfish families Loricariidae, Astroblepidae and Mochokidae display the most dramatically adapted morphologies. Loricariidae and Astroblepidae, living in the Neotropical freshwaters, exclusively contain suckermouth catfish species, and their anatomy and head kinematics have already been studied into detail. Among Mochokidae, living in the tropical freshwaters of Africa, only the chiloglanidine subfamily has a sucker mouth, and occupies similar niches in Africa as both Neotropical families do in South America. Having derived from relatively unrelated catfish ancestors, their anatomy is poorly known, and the nature of their scraping and station-holding capabilities is not known at all. This paper provides details on the chiloglanidine head anatomy and function (relating their anatomy to that of the non-suckermouth Mochokidae), and compares this Afrotropical suckermouth taxon with both Neotropical suckermouth families. It identifies both convergences and differing anatomical and kinematic solutions to the same key needs of food-scraping and station-holding suckermouth fishes. Chiloglanidine mochokids differ from both Neotropical families in having less mobile jaws, with an upper jaw assisting more in station-holding than in feeding. They share the highly mobile lower lip with both Neotropical taxa, although the configuration of the intermandibular/protractor hyoidei muscle system, changing the volume of the sucker-disc cavity, differs in all three taxa. Chiloglanidines have a single, posterior inflow opening into this cavity, whereas Loricariidae have two lateral openings, and Astroblepidae have none, using an opercular incurrent opening instead. The chiloglanidine buccal valve system consists of two passive valves, as in Astroblepidae. Although less diverse in number of genera and species, this Afrotropical suckermouth taxon possesses the anatomical and kinematic key elements allowing a successful occupation of a niche similar to the one found in the Loricariidae + Astroblepidae clade.

Functional morphology of the Andean climbing catfishes (Astroblepidae, Siluriformes): alternative ways of respiration, adhesion, and locomotion using the mouth.

Astroblepidae or "climbing catfishes" encompass a single genus of species living in high altitude rivers in the Andes of South America. They are characterized by a specialized head morphology closely resembling their better known, widely radiated sister family Loricariidae, or armored suckermouth catfishes. Existent data show that even though both families share important traits, there are some striking differences as well. Albeit poorly known, Astroblepus species possess a duplicated gill opening, and have the ability to climb vertical rocks or waterfalls. In this study, morphological and kinematic data are combined to yield insights into the functions of the mobile elements of the astroblepid head, and to compare head morphology and biomechanics with those of Loricariidae. We found that, even though there is substantial similarity in head structure of both families, there are major differences in functionally important structures. These include a different lower lip muscle configuration, an alternative oral valve system, and an incurrent gill opening only found in astroblepids. Kinematic analyses confirm that the astroblepid suckermouth, freed from its inhalatory function, offers advantages for climbing in the high-altitude environment, and is used alternately with the extremely mobile pelvic girdle, in a crawling, nonundulatory motion.

Description of Notoglanidium pembetadi new species (Siluriformes: Claroteidae) from the Kouilou-Niari River, Republic of the Congo.

A new claroteid catfish, Notoglanidium peinbetadi, is described from the Kouilou-Niari River basin (Republic of the Congo). This species can be distinguished from all other currently known Notoglanidium species as well as from the very similar Anaspidoglanis boutchangai, to which two specimens have erroneously been attributed in the past, by the following combination of characters: II, 10-13 dorsal-fin rays; long dorsal-fin base (21.1-24.4% SL); short predorsal distance (30.1-33.5% SL); and short prepelvic distance (40.1-45.4% SL).

Revision of Notoglanidium and related genera (Siluriformes: Claroteidae) based on morphology and osteology.

Apart from the well-demarcated genera Auchenoglanis and Parauchenoglanis, Auchenoglanidinae, one of the two sub-families of the African catfish family Claroteidae, suffers from poor resolution at the generic level. For the remaining genera, Notoglanidium, Liauchenoglanis, Platyglanis and Anaspidoglanis, generic discriminations are rudimentary. In addition, several included species are poorly defined and barely represented in scientific collections. Until now, no study has included morphological data for all currently known species, and for many species osteological data were non-existent. Molecular data for most species are lacking as well. Here, a comprehensive account of the morphology and osteology is given of all species included in these four genera. Using computed tomography (CT scanning) as well as clearing and staining, osteological characters were combined with biometric, meristic and other morphological data to revise the status of these genera and included species. Morphological and osteological data, submitted to a phylogenetic analysis, agree with metric and meristic data that all Liauchenoglanis, Platyglanis and Anaspidoglanis cannot be discerned from Notoglanidium; their genus and type species descriptions fail to be distinguished from Notoglanidium. Here their synonymy is proposed, resulting in a single valid genus, including nine species of which the validity is confirmed. A key to the genus and included species, as well as a diagnosis and description for each of them, are provided.

Soft dentin results in unique flexible teeth in scraping catfishes.

Teeth are generally used for actions in which they experience mainly compressive forces acting toward the base. The ordered tooth enamel(oid) and dentin structures contribute to the high compressive strength but also to the minor shear and tensile strengths. Some vertebrates, however, use their teeth for scraping, with teeth experiencing forces directed mostly normal to their long axis. Some scraping suckermouth catfishes (Loricariidae) even appear to have flexible teeth, which have not been found in any other vertebrate taxon. Considering the mineralized nature of tooth tissues, the notion of flexible teeth seems paradoxical. We studied teeth of five species, testing and measuring tooth flexibility, and investigating tooth (micro)structure using transmission electron microscopy, staining, computed tomography scanning, and scanning electron microscopy-energy-dispersive spectrometry. We quantified the extreme bending capacity of single teeth (up to 180°) and show that reorganizations of the tooth (micro)structure and extreme hypomineralization of the dentin are adaptations preventing breaking by allowing flexibility. Tooth shape and internal structure appear to be optimized for bending in one direction, which is expected to occur frequently when feeding (scraping) under natural conditions. Not all loricariid catfishes possess flexible teeth, with the trait potentially having evolved more than once. Flexible teeth surely rank among the most extreme evolutionary novelties in known mineralized biological materials and might yield a better understanding of the processes of dentin formation and (hypo)mineralization in vertebrates, including humans.

Suckermouth armored catfish resolve the paradox of simultaneous respiration and suction attachment: a kinematic study of Pterygoplichthys disjunctivus.

Suckermouth armored catfishes (Loricariidae) use their suckermouth for inspiration, feeding, and attachment to substrates. The sucker consists of a pre-valvular cavity, formed by a modified lip disc, and is separated from the larger post-valvular buccal cavity by a muscular oral valve. The combination of respiration and suction attachment seems paradoxal, as a properly functioning suction device would need a sucker without leakage (yet inspiration must occur via the sucker), and continuous subambient pressure in the sucker cavity (even during expiratory mouth floor elevation). In the loricariid Pterygoplichthys disjunctivus, the anatomy of the suckermouth structures was examined, and a kinematic analysis was performed to acquire insights into how respiration and attachment are combined. High-speed external and X-ray recordings show that suckermouth attachment influences respiratory parameters such as decreasing excursion amplitudes of mouth floor elements, and the way water enters the mouth via furrows in the lip disc. Respiration, however, continues during attachment and is not blocked. Our data show that the muscular oral valve actively separates the post-valvular buccal cavity from the pre-valvular sucker cavity. Volume changes of this pre-valvular cavity are opposite to those of the post-valvular cavity and assure sucker function even during expiration. These volume changes are caused by movements of the lower lip, the lower jaws, and the oral valve. The lateral inflow furrow openings, controlled by the maxillary barbels, can occur unilaterally. Morphological and kinematic data also show that the opercle is anatomically and functionally decoupled from the gill opening.

Development of the osteocranium in Corydoras aeneus (Gill, 1858) Callichthyidae, Siluriformes.

Development in the osteocranium of Corydoras aeneus was studied based on 48 cleared and stained specimens and 10 series of serial sections. Development overall follows the general trends observed in siluriform development, with ossifications appearing as a response to functional demands. Early development of the skull occurs in two distinct phases. In a first phase, several new bony elements, all of dermal origin and related to feeding, appear shortly after yolk depletion (4.4 mm SL). Between 5 and 8 mm SL, developmental priorities seem to shift to size increase of the cartilaginous skull and no new bony elements appear. Finally, a second phase of osteogenesis occurs from 8 to 18 mm SL, in which all remaining dermal and perichondral bones appear.

Kinematics of benthic suction feeding in Callichthyidae and Mochokidae, with functional implications for the evolution of food scraping in catfishes.

Food scraping has independently evolved twice from suction feeding in the evolution of catfishes: within neotropical Loricarioidea and paleotropical Mochokidae. To gain insight in the evolutionary transitions associated with the evolution towards scraping, we analyzed prey capture kinematics in two species of benthic suction feeders which belong to taxa that are closely related to the scraper lineages (respectively, Corydoras splendens and Synodontis multipunctatus), and compared it to prey capture in a more distantly related, generalist suction feeder (Clarias gariepinus). Simultaneous ventral and lateral view high-speed videos were recorded to quantify the movements of the lower jaw, hyoid, pectoral girdle and neurocranium. Additionally, ellipse modeling was applied to relate head shape differences to buccal expansion kinematics. Similarly to what has been observed in scrapers, rotations of the neurocranium are minimal in the benthic suction feeders, and may consequently have facilitated the evolution of a scraping feeding mechanism. The hypothesis that fish with a more laterally compressed head rely more heavily on lateral expansion of the buccal cavity to generate suction, was confirmed in our sample of catfish species. Since an important contribution of lateral expansion of the head to suction may avoid the need for a strong, ventral depression of the mouth floor during feeding, we hypothesized that this may have allowed a closer association with the substrate in the ancestors of scrapers. However, our hypothesis was not supported by an ancestral state reconstruction, which suggests that scraping probably evolved from sub-terminal mouthed ancestors with dorsoventrally flattened heads.

Extensive jaw mobility in suckermouth armored catfishes (Loricariidae): a morphological and kinematic analysis of substrate scraping mode of feeding.

Loricariidae, or suckermouth armored catfishes, possess upper and lower jaws that are ventrally oriented and that bear teeth that touch the substrate from which algae and other food items are scraped. The ventral orientation and the highly specialized morphology of the jaws, characterized by protrusible upper jaws and left-right decoupled lower jaws, are observed in Pterygoplichthys disjunctivus, the species investigated here. Kinematic data of the scraping feeding movements, obtained by external high-speed and x-ray recordings, are used to quantify jaw movement, especially to test for upper jaw mobility and versatility during substrate scraping. Our results show that the mobility of the jaws is indeed high compared with what is standard for catfishes. The upper jaw's ability to perform a substantial degree of rostrocaudal movement is quite unique for catfishes. The ventromedially oriented lower jaws, with the teeth and the coronoid process at opposite sides, display an extensive mobility: they rotate around the suspensorial articulation and around their longitudinal axis, resulting in an extended scraping movement and thereby covering a large surface area. The lower jaws also show a left-right asymmetry in their movements during scraping. Thus, our results suggest that the extreme morphological specializations of the jaws in loricariid catfishes are linked to an increased mobility and functional versatility, allowing these animals to efficiently scrape algae from substrates with irregular surfaces.

Morphology and development of teeth and epidermal brushes in loricariid catfishes.

Loricariidae or suckermouth armored catfishes are one of several aquatic taxa feeding on epilithic and epiphytic algae. Their upper and lower jaws bear exquisitely curved teeth, which usually are asymmetrically bicuspid. The enlarged lower lip carries papillae with keratinous unicellular epidermal brushes or unculi. Teeth, and probably unculi too, assist in scraping food off substrates. Their morphology, growth, and replacement is examined and compared among several loricariid species, using cleared and stained specimens, serial sections, and SEM. Apart from the general tooth form and crown shape, the anterior layer of soft tissue on the lower shaft region, present in several species, appears to be a specialization for enhancing the mobility of individual teeth when scraping on uneven surfaces. During early ontogeny, a transition from simple conical to mature tooth occurs. The first unculi appear together with the first teeth carrying a bicuspid crown, 2 days after the first exogenous feeding, but synchronous with the complete resorption of the yolk sac.

Development of the osteocranium in the suckermouth armored catfish Ancistrus cf. triradiatus (Loricariidae, siluriformes).

The development of the osteocranium of the suckermouth armored catfish Ancistrus cf. triradiatus is described based on specimens ranging from prehatching stages to juvenile stages where the osteocranium is more or less fully formed. The first bony elements that arise are the opercle, jaws, and lateralmost branchiostegal rays, as well as the basioccipital and parasphenoid in the skull floor. The supracleithrum and the membranous and perichondral pterotic components form one large, double-layered skull bone during ontogeny, without clear evidence of the involvement of a supratemporal. The Baudelot's ligament ossifies from two sides, i.e., from the basioccipital medially and the supracleithrum laterally. The lower jaw consists of a dentary, mentomeckelian, and angulo-articular, which all soon fuse. The parurohyal, formed by the fusion of a ventral sesamoid bone and a dorsal cartilage element associated with the first basibranchial, is pierced by a vein, unlike in some other siluriforms. The interhyal cartilage disappears during ontogeny; medially of it, a small sesamoid bone appears in a ligament. The largest, canal-bearing cheek plate is not homologous to the interopercle. The results of the present research, with emphasis on bone formations and homologies, are compared with studies on related catfishes.

The erectile cheek-spine apparatus in the bristlenose catfish Ancistrus (Loricariidae, Siluriformes), and its relation to the formation of a secondary skull roof.

In the South American catfish family Loricariidae, the opercle has been decoupled from the lower jaw, and has also lost its function in expiration. While many loricariid species have a small and slightly mobile opercle with reduced opercular musculature, within the hypostomine subfamily a novel opercular mechanism has developed that erects a tuft of enlarged odontodes anterior to the opercle. This defensive mechanism is examined in Ancistrus cf. triradiatus. The opercle has a prominent anterior process and the orientation of the reinforced articulation hinge to the hyomandibular bone has shifted. The opercular musculature is well developed, with a hypertrophied dilatator operculi that extends deep inside the skull roof bones and toward the midline, over the brain, but below the superficial skull roof. Hence the frontal, sphenotic, parieto-supraoccipital and compound pterotic bones consist of a dorsal, superficial part and a deeper part separating the brain from the muscle: two functional skull roofs are thus formed. The impact on the path of the cranial sensory canals is substantial, moving canals away from the skull surface. Hypertrophy of cranial muscles is known from many teleosts, but the invasion of such large muscles into the skull, which is drastically modified and literally hollowed out, has never been described before. These cranial modifications are greater in males than in females, related to the territorial behavior of the former, in which the erectile spines are usually used.

Development of the chondrocranium in the suckermouth armored catfish Ancistrus cf. triradiatus (Loricariidae, Siluriformes).

The chondrocranium of the suckermouth armored catfish Ancistrus cf. triradiatus was studied. Its development is described based on specimens ranging from small prehatching stages with no cartilage visible, to larger posthatching stages where the chondrocranium is reducing. Cleared and stained specimens, as well as serial sections, revealed a cartilaginous skeleton with many features common for Siluriformes, yet several aspects of A. cf. triradiatus are not seen as such in other catfishes, or to a lesser extent. The skull is platybasic, but the acrochordal cartilage is very small and variably present, leaving the notochord protruding into the hypophyseal fenestra in the earlier stages. The ethmoid region is slender, with a rudimentary solum nasi. A lateral commissure and myodomes are present. The larger posterior myodome is roofed by a prootic bridge. The maxillary barbel is supported by a conspicuous cartilaginous rod from early prehatching stages. The ceratohyal has four prominent lateral processes. Infrapharyngobranchials I-II do not develop. During ontogeny, the skull lengthens, with an elongated ethmoid, pointing ventrally, and a long and bar-shaped hyosymplectic-pterygoquadrate plate. Meckel's cartilages point medially instead of rostrally.