Measurement of jaw viscoelasticity in newborn and adult lesser spotted dogfish Scyliorhinus canicula (L., 1758)

Jaw viscoelasticity of newborn and adult lesser spotted dogfish Scyliorhinus canicula was measured using dynamic mechanical analysis. Jaws of newborn animals were significantly more viscoelastic than those of adults. As a result, newborn lesser spotted dogfish might be unable to consume hard prey items, unlike adults.     

The polarization of inner ear ciliary bundles from a scorpaeniform fish

The polarization of ultrastructural ciliary bundles from hair cells in the inner ear of the sea scorpion Taurulus bubalis was studied using a scanning electron microscope, revealing arrays of ciliary bundles with diverse orientations on each of the sensory epithelia. Members of this order are known to produce sound, though results of this study show no significant variation from the standard receptor patterns found in the hearing system of many silent marine teleosts. This is the first time that the ultrastructure of T. bubalis has been studied, and this work presents a new set of polarization patterns, which provide anatomical information important in understanding electrophysiological aspects of fish hearing from an ecological perspective.     

Fatty acid composition and lipoxygenase metabolism in blood cells of the lesser spotted dogfish, Scyliorhinus canicula.

1. The fatty acid composition of erythrocytes and leucocytes of the elasmobranch, Scyliorhinus canicula, was determined so as to indicate substrate availability for eicosanoid formation. 2. Leucocytes showed a greater degree of fatty acid unsaturation than the erythrocytes, with particularly high levels of docosahexaenoic acid (22:6,n-3). 3. The major eicosanoid precursors, arachidonic acid (20:4,n-6) and eicosapentaenoic acid (20:5,n-3), represented 13.9% and 5.2% of the total fatty acid, respectively, in erythrocytes compared with 10.7% and 6% in leucocytes. 4. Whole blood and isolated leucocytes were stimulated with calcium ionophore, A23187 and the resulting lipoxygenase products separated by reverse phase high performance liquid chromatography. 5. The main lipoxygenase products formed were 6-trans-leukotriene B4, 6-trans-12-epi-leukotriene B4, 5(S),6(R) dihydroxyeicosatetraenoic acid and 5- and 15-hydroxyeicosatetraenoic acid. 6. No leukotriene B4, leukotriene B5, or lipoxins were detected.     

On the diel rhythms in metabolism and activity of post-hatching lesser spotted dogfish, Scyliorhinus canicula

The diel rhythms in metabolic rate ( M_R ) and activity level ( A_L ) were measured for single post-hatching dogfish (weight range, 2.76–10.61 g) at 15° C by the indirect calorimetric method of rate of oxygen consumption ( V O₂) and by video-observation respectively, over a period of 72 b. The mean VO ₂ increased from 62.0 (s.e. 2.9) mg O₂ kg⁻¹ h⁻¹ in the daylight hours to 85.5 (s.e. 3.1) mg O₂ kg⁻¹ h⁻¹ during the dark (light regíme, 12 h L: 12 h D). The simultaneous measurement of A _L also showed mean night elevation from 0.6 (s.e. 0.2) min h⁻¹ in the light phase to 14.5 (s.e. 1.6) min h⁻¹ during the darkness. Bimodal nocturnal activity (BNA) was exhibited by the post-hatching dogfish within the 12 h dark period, with V O₂ increasing from 71.4 (s.e. 2.8) mg O₂ kg⁻¹ h⁻¹ before 01.00 hours to 99.5 (s.e. 4.2) mg O₂ kg⁻¹ h⁻¹ after 01.00 hours. Similarly, A _L also increased from 8.9 (s.e. I.7)min h⁻¹ before 01.00 hours to 21.1 (s.e. 2.8) min h⁻¹ after 01.00 hours. The importance of the results presented to the natural behavioural ecology of the hatching dogfish are discussed.     

Ciliated cells on the surface of embryos of Scyliorhinus canicula

Monociliated surface cells were found for the first time in an elasmobranch and were common on embryos of Scyliorhinus canicula . They occurred mainly near the anterior end, until Ballard stage 30, but disappeared soon thereafter. The functions of surface ciliated cells suggested in other embryos ( e.g. surface currents in lungfishes and amphibians, and determination of left–right patterns in amniotes) seem unlikely in elasmobranchs.     

Evidence for multiple,developmentally regulated isoforms of Ptprq on hair cells of the inner ear

Ptprq is a receptor‐like inositol lipid phosphatase associated with the shaft connectors of hair bundles. Three lines of evidence suggest Ptprq is a chondroitin sulfate proteoglycan: (1) chondroitinase ABC treatment causes a loss of the ruthenium‐red reactive, electron‐dense particles associated with shaft connectors, (2) chondroitinase ABC causes an increase in the electrophoretic mobility of Ptprq, and (3) hair bundles in the developing inner ear of wild‐type mice, but not those of Ptprq^?/? mice, react with monoclonal antibody (mAb) 473‐HD, an IgM that recognizes the dermatan‐sulfate‐dependent epitope DSD1. Two lines of evidence indicate that there may be multiple isoforms of Ptprq expressed in hair bundles. First, although Ptprq is expressed throughout the lifetime of most hair cells, hair bundles in the mouse and chick inner ear only express the DSD1 epitope transiently during development. Second, mAb H10, a novel mAb that recognizes an epitope common to several avian inner‐ear proteins including Ptprq, only stains mature hair bundles in the extrastriolar regions of the vestibular maculae. MAb H10 does not stain mature hair bundles in the striolar regions of the maculae or in the basilar papilla, nor does it stain immature hair bundles in any organ. Three distinct, developmentally regulated isoforms of Ptprq may therefore be expressed on hair bundles of the chick inner ear. Hair bundles in the mature chick ear that do not express the H10 epitope have longer shaft connectors than those that do, indicating the presence or absence of the H10 epitope on Ptprq may modulate the spacing of stereocilia. © 2010 Wiley Periodicals, Inc. Develop Neurobiol 71: 129‐141, 2011     

The annual reproductive cycle of the female lesser spotted dogfish, Scyliorhinus canicula L., and its endocrine control

Various aspects of the reproductive physiology of the female lesser spotted dogfish, Scyliorhinus canicula L., demonstrate marked annual cycles. Pituitary ventral lobe gonadotrophin content and plasma oestradiol and testosterone levels rise through the autumn as the ovary recrudesces and the percentage of fish bearing eggs in the oviducts increases. The pituitary gonadotrophin and plasma sex steroid levels remain elevated throughout the winter but decrease during the spring, two months prior to a fall in both the gonadosomatic index (GSI) and the percentage of fish with eggs in their oviducts. The lowest gonadotrophin and sex steroid levels are found in early summer, again preceding the minimum GSI and rate of egg-laying by two or three months. Some of these cycles are pronounced, the pituitary gonadotrophin levels varying by more than 100-fold, and the oestradiol levels by more than 10-fold, throughout the year. It is concluded that the female dogfish has a very extended breeding season, though the peak frequency of egg-laying occurs in the winter and spring.     

An ultrastructural and immunocytochemical study on the lymphomyeloid tissue in the embryonic kidney of the dogfish, Scyliorhinus canicula

The central role of the kidney during early development in the dogfish, Scyliorhinus canicula , is haemopoietic. The kidney is the first tissue to become lymphomyeloid and lymphocytes and developing granulocytes are found in the interstitial tissue surrounding the renal tubules. The lymphomyeloid role of the kidney is transient and does not persist after hatching. The kidney is a major immunoglobulin (Ig) containing tissue during early development as evidenced by increasing numbers of Ig–positive cells.     

Food and feeding habits of the lesser spotted dogfish, Scyliorhinus canicula (L.), in Isle of Man waters

Stomach contents of the lesser spotted dogfish, Scyliorhinus canicula , from Isle of Man waters were investigated. Dogfish fed opportunistically on a wide range of macrobenthic fauna with hermit crabs, cockles and whelks the dominant prey. Items of secondary importance included various crabs, callianassid shrimps, bivalve molluscs, holothurians, polychaetes and, when locally abundant, herring, Clupea harengus . Dogfish predated more heavily on molluscs than reported in previous studies, reflecting the abundance of molluscs in the associated benthic community.
Composition of the diet altered gradually with dogfish size, reliance on small crustaceans declined and consumption of hermit crabs and molluscs increased with growth. Seasonal changes in the composition of the diet were observed. Feeding intensity was greatest during summer, related in part to increased prey availability, and was least during autumn.     

The effect of body size on the standard metabolic rate of the lesser spotted dogfish

Standard metabolic rates were measured as the rate of oxygen consumption in 33 Scyliorhinus canicula , ranging in weight from 3–929 g. The amount of oxygen consumed per hour (Vo₂) changed predictably with body size according to the relationship Vo₂=0–104 ^W0.855, where W represents fish weight in g. On a weight specific basis, the level of standard metabolism in juvenile dogfish (5 g) was nearly double that measured in adults (500 g).     

Dark hair cells in the inner ear of the rainbow trout. A study of the influence of different fixation methods

The occurrence of dark staining cells in different tissues has been suggested to be artefactual and caused during the fixation process. In inner ear sensory epithelia, dark hair cells (DHC) have been suggested to be apoptotic cells. We have examined whether dark cells represent dying cells or whether they are the results of fixation artefacts. The effects of buffer osmolarity and different fixation methods on the incidence of dark hair cells in the inner ear macula sacculi of the rainbow trout (Oncorhynchus mykiss) were investigated by light and electron microscopy. Glutaraldehyde in phosphate buffer with osmolarities of 0, 135, 225, 425, and 560 mosmol were used for fixation by immersion. For comparison, fixation by vascular perfusion as well as the effects of mechanical injury and delayed fixation were studied. DHC were found in all examined saccular maculae except for the delayed fixation protocol where almost all the sensory cells were lost. The number of DHC accounted for 2.5–12.9‰ of the sensory cells. Neither the buffer osmolarity nor the fixation method had significant effects on the frequencies of DHC. Mitotic cell division events were seen exclusively in the apical cell strata of the sensory epithelium. The DHC are suggested to be associated with apoptosis rather than fixation artefacts.     

FGF信号通路在内耳发育调控和毛细胞再生中的作用

内耳是感受听觉和平衡觉的复杂器官。在内耳发育过程中,成纤维生长因子(fibroblast growth factor, FGF)信号通路参与了听基板的诱导、螺旋神经节(statoacoustic ganglion, SAG)的发育以及Corti器感觉上皮的分化。FGF信号开启了内耳早期发育的基因调控网络,诱导前基板区域以及听基板的形成。正常表达的FGF信号分子可促进听囊腹侧成神经细胞的特化,但成熟SAG神经元释放的过量FGF5可抑制此过程,形成负反馈环路使SAG在稳定状态下发育。FGF20在Notch信号通路的调控下参与了前感觉上皮区域向毛细胞和支持细胞的分化过程,而内毛细胞分泌的FGF8可调控局部支持细胞分化为柱细胞。人类FGF信号通路异常可导致多种耳聋相关遗传病。此外,FGF信号通路在低等脊椎动物毛细胞自发再生以及干细胞向内耳毛细胞诱导过程中都起到了关键作用。本文综述了FGF信号通路在内耳发育调控以及毛细胞再生中的作用及其相关研究进展,以期为毛细胞再生中FGF信号通路调控机制的阐明奠定理论基础。     

Epithelial-mesenchymal transitions in the developing heart of the dogfish (Scyliorhinus canicula). A scanning electron microscopic study

An epithelial-mesenchymal transition is involved in two main morphogenetic events of cardiac morphogenesis, namely the differentiation of the valvuloseptal tissue from the endocardial endothelium, and the formation of subepicardial mesenchyme from the epicardial mesothelium. We have proposed that the dogfish ( Scyliorhinus canicula ) is a suitable model for the study of basic processes of cardiac morphogenesis in vertebrates, since the heart of this primitive fish probably outlines the original bauplan of the vertebrate heart. In order to study in this model the endocardial and epicardial epithelial-mesenchymal transition under scanning electron microscopy, we have used a technique of paraffin-embedding, partial sectioning, dewaxing and critical-point drying. Our results showed: 1) A centrifugal pattern of epicardial development from the atrioventricular groove to the sinus venosus and conus arteriosus; 2) A close spatial and temporal relationship between the endocardial and epicardial epithelial-mesenchymal transition, although the transformation of the endocardium starts earlier and ends later the epicardial transformation; 3) A complex arrangement of the fibrous extracellular matrix which is established prior to the migration of the mesenchymal cells. Subepicardial, but not subendothelial mesenchymal cells, coalesce in unicellular or pluricellular ring-like structures that probably are related to the origin of the cardiac vessels.     

Characterization of a chymosin-like pepsin from the dogfish Scyliorhinus canicula

1. Pepsin II extracted from the gastric mucosa of Scyliorhinus canicula has been characterized and compared to calf chymosin. 2. The kcat and Km of the dogfish enzyme for the synthetic hexapeptide Leu-Ser-Phe(NO2)-Nle-Ala-Leu-OMe have been determined. The kcat/Km ratio is close to that of calf chymosin. Its milk-clotting efficiency is however 21-fold lower than that of calf chymosin. 3. The proteolytic activity against haemoglobin is optimal at pH 2.5. It clots the milk up to pH 6.8. 4. The dogfish pepsin II shows relatively better activity at low temperatures than calf chymosin.     

The developmental expression of foxl2 in the dogfish Scyliorhinus canicula

The FoxL2 genes are a subfamily of the Fox (forkhead box) gene family. FOXL2 is mutated in the disorder Blepharophimosis, Ptosis, and Epicanthus Inversus Syndrome (BPES), which is characterized by eyelid malformations, and Premature Ovarian Failure (POF). In the mouse expression is seen in the perioptic mesenchyme, developing eyelids, ovary and pituitary. We have isolated a foxl2 cDNA from the dogfish Scyliorhinus canicula (also known as the lesser spotted catshark), allowing the characterisation of this gene's sequence and expression from a lineage that diverged early in the evolution of gnathostomes. Molecular phylogenetic analysis strongly grouped this sequence with the gnathostomes within the FoxL2 subfamily. We demonstrate the early expression of Scyliorhinus canicula foxl2 in the mandibular head mesoderm and later in continuous populations of mandibular arch cells and mandibular head mesenchyme cells around the developing pituitary. As development proceeds expression decreases in the mesenchyme of the head but is seen in the mesenchyme around the eye and later in the developing eyelids. Additionally expression is seen in regions of pharyngeal arch mesoderm and in ectoderm from which gill buds will form. This expression is maintained in the developing and elongating gill buds. Thus, S. canicula foxl2 is a marker for the mandibular mesoderm and gill buds and its expression is conserved in the perioptic mesenchyme, developing eyelids and pituitary.     

Structure and expression of three Emx genes in the dogfish Scyliorhinus canicula: functional and evolutionary implications

We report the characterization of three Emx genes in a chondrichthyan, the dogfish Scyliorhinus canicula. Comparisons of these genes with their osteichthyan counterparts indicate that the gnathostome Emx genes belong to three distinct orthology classes, each containing one of the dogfish genes and either the tetrapod Emx1 genes (Emx1 class), the osteichthyan Emx2 genes (Emx2 class) or the zebrafish Emx1 gene (Emx3 class). While the three classes could be retrieved from the pufferfish genome data, no indication of an Emx3-related gene in tetrapods could be found in the databases, suggesting that this class may have been lost in this taxon. Expression pattern comparisons of the three dogfish Emx genes and their osteichthyan counterparts indicate that not only telencephalic, but also diencephalic Emx expression territories are highly conserved among gnathostomes. In particular, all gnathostomes share an early, dynamic phase of Emx expression, spanning presumptive dorsal diencephalic territories, which involves Emx3 in the dogfish, but another orthology class, Emx2, in tetrapods. In addition, the dogfish Emx2 gene shows a highly specific expression domain in the cephalic paraxial mesoderm from the end of gastrulation and throughout neurulation, which suggests a role in the segmentation of the cephalic mesoderm.     

In vitro differentiation of mouse embryonic stem cells into inner ear hair cell-like cells using stromal cell conditioned medium

Hearing loss is mainly caused by loss of sensory hair cells (HCs) in the organ of Corti or cochlea. Although embryonic stem (ES) cells are a promising source for cell therapy, little is known about the efficient generation of HC-like cells from ES cells. In the present study, we developed a single-medium culture method for growing embryoid bodies (EBs), in which conditioned medium (CM) from cultures of ST2 stromal cells (ST2-CM) was used for 14-day cultures of 4-day EBs. At the end of the 14-day cultures, up to 20% of the cells in EB outgrowths expressed HC-related markers, including Math1 (also known as Atoh1), myosin6, myosin7a, calretinin, α9AchR and Brn3c (also known as Pou4f3), and also showed formation of stereocilia-like structures. Further, we found that these cells were incorporated into the developing inner ear after transplantation into chick embryos. The present inner ear HC induction method using ST2-CM (HIST2 method) is quite simple and highly efficient to obtain ES-derived HC-like cells with a relatively short cultivation time.     

Sox2在内耳发育中的作用

转录因子Sox2是Sox基因家族的成员之一,由于它在早期胚胎发生、神经分化和内耳发育等多种重要的发育事件中都起着关键的作用,从而引起了越来越广泛的关注。哺乳动物的内耳主要由6个形态上和功能上不同的感觉区组成,这些区域对声音和前庭信息的传导是必需的,在这些区域的发育过程中,Sox2基因是内耳细胞早期发育所必需的基因。该文就Sox2在内耳发育中的作用作一综述。     

Dissociation and identification of intact seminiferous lobules from the testis of the dogfish (Scyliorhinus canicula)

Summary An enzymatic method was developed to obtain intact seminiferous lobules from the testis of the dogfish (Scyliorhinus canicula L.). The freshly isolated lobules were then identified by use of a transillumination technique. Testes from mature dogfish were collected and transverse sections incubated with a mixture of collagenase (0.025%) + pronase (0.08%) at 4° C overnight. Dissociation of the tissue was achieved by mechanical agitation in a calcium and magnesium-free buffer (pH 7.8; 870 mOsm) at room temperature, for 30 min. Based on differences in light absorption and size as well as on comparison with the corresponding histological and ultrastructural cell composition, the seminiferous lobules were identified and classified according to the stages of spermatogenesis of Mellinger (1965). The characteristic changes take place in the size and surface of the lobules and are mainly due to differences in the arrangement of the germ cells, in their number, and in the light absorption characteristics of their nuclei. The combination of the procedures of isolation and transillumination of the dogfish seminiferous lobules, in native condition, offers an original method for the study of germ cell-Sertoli cell interactions in the non-mammalian vertebrate.