Differentiation and growth of kype skeletal tissues in anadromous male Atlantic salmon (Salmo salar)

The re-initiation of bone development in adult starving Atlantic salmon (Salmo salar) during their energetically expensive upstream migration is remarkable and deserves closer examination. Dramatic alterations of the skull bones and teeth, most prominently, the development of a kype in males, are widely known but little studied or understood. We describe the microstructure and the cellular processes involved in the formation of the skeletal tissues of the kype. Fresh bone material, obtained from animals migrating upstream was subjected to radiological, histological or histochemical analysis. We show that the kype is, in part, composed of rapidly growing skeletal needles arising at the tip of the dentary. Proximally, the needles anastomose into a spongiosa-like meshwork which retains connective tissue inside bone marrow spaces. Ventrally, the needles blend into Sharpey fiber bone. Skeletal needles and Sharpey fiber bone can be distinguished from the compact bone of the dentary by radiography. Rapid formation of the skeleton of the kype is demonstrated by the presence of numerous osteoblasts, a broad distal osteoid zone, and the appearance of proteoglycans at the growth zone. The mode of bone formation in anadromous males can be described as 'making bone as fast as possible and with as little material as possible'. Unlike the normal compact bone of the dentary, the new skeletal tissue contains chondrocytes and cartilaginous extracellular matrix. Formation of the skeleton of the kype resembles antler development in deer (a form of regeneration), or hyperostotic bone formation in other teleost fishes, rather than periosteal bone growth. The type of boneformation may be understandable in the light of the animals' starvation and the energetic costs of upstream migration. However, the structured and regulated mode of bone formation suggests that the skeleton of the kype has functional relevance and is not a by-product of hormonal alterations or change of habitat

Periderm invasion contributes to epithelial formation in the teleost pharynx

The gnathostome pharyngeal cavity functions in food transport and respiration. In amniotes the mouth and nares are the only channels allowing direct contact between internal and external epithelia. In teleost fish, gill slits arise through opening of endodermal pouches and connect the pharynx to the exterior. Using transgenic zebrafish lines, cell tracing, live imaging and different markers, we investigated if pharyngeal openings enable epithelial invasion and how this modifies the pharyngeal epithelium. We conclude that in zebrafish the pharyngeal endoderm becomes overlain by cells with a peridermal phenotype. In a wave starting from pouch 2, peridermal cells from the outer skin layer invade the successive pouches until halfway their depth. Here the peridermal cells connect to a population of cells inside the pharyngeal cavity that express periderm markers, yet do not invade from outside. The latter population expands along the midline from anterior to posterior until the esophagus-gut boundary. Together, our results show a novel role for the periderm as an internal epithelium becomes adapted to function as an external surface.Agência financiadora Ghent University Research Fund - BOF24J2015001401 Cancer Prevention Research Institute of Texas - RR140077info:eu-repo/semantics/publishedVersio

Analysis of a short tail type in farmed Atlantic salmon (Salmo salar)

The short tail phenotype represents one of the main causes for downgrading of farmed Atlantic salmon (Salmo salar) at slaughterhouses. Prevalence of short tail is variable and the aetiology is suspected to be multi-factorial. Risk factors have been identified but descriptions of the aetiology and the pathology of the condition are still rare. In the current study, a radiological and histological analysis of short tails has been performed, examining six normal and six downgraded individuals from a slaughterhouse in southern Norway. In the short tail phenotype, vertebral bodies were shifted and bent at the contact zone of adjacent vertebral bodies. Changes either affected the entire spine or were located at the medial caudal-spine. While the internal bone structure of the vertebrae was similar in deformed and non-deformed animals, a lack of intervertebral space apparently caused a shortening of the vertebral column and corresponded to an elevated condition factor in deformed individuals. Histological analysis revealed different degrees of proliferation of cartilaginous tissues, which replaced the intervertebral notochord tissue. The displacement of adjacent vertebral bodies and the development of cartilage in between vertebral bodies suggest mechanical forces as a possible cause for the observed deformations, since mechanically-induced overload and a subsequent direct contact of bones are factors that can stimulate heterotopic cartilage development and pseudoarthrosi

Patterns of tooth replacement in osteichthyans: variations on a theme

Nonmammalian tooth-bearing vertebrates usually replace their teeth throughout life. Much about how a replacement pattern is generated has been learned from zebrafish. However, to understand general mechanisms of tooth replacement, advantage can be taken from studying other, “nonmodel” species. We have mapped the patterns of tooth replacement in widely divergent aquatic osteichthyans using 2D charts, in which one axis is time, the other linear spacing along the tooth row. New teeth that are generated simultaneously are considered part of the same odontogenic wave. Using this approach, it appears that a similar, general pattern underlies very distinctive dentitions in distantly related species. A simple shift in spacing of odontogenic waves, or in distance between subsequent tooth positions along a row (or both), can produce dramatically different dentitions between life stages within a species, or between closely related species. Examples will be presented from salmonids, cyprinids, and cichlids. Our observations suggest that lines linking subsequent positions may have more biological significance than replacement waves (usually linking alternate positions), often used to explain the generation of patterns. The presence of a general pattern raises questions about common control mechanisms. There is now increasing evidence, at least for the zebrafish, to support a role for stem cells in continuous tooth renewal and control of replacement patterns

Vertebral column adaptations in juvenile Atlantic salmon Salmo salar, L. as a response to dietary phosphorus

Deficiency in dietary phosphorus (P) is considered as a nutritional risk factor for the development of vertebral column deformities in farmed Atlantic salmon Salmo salar, L. This mono-factorial study examines how 11-week deficiency and excess of dietary P influence the structure and microstructure of the vertebral bodies in juvenile, freshwater stages of Atlantic salmon. Animals were fed continuously with three diets containing different levels of total P (tP) and soluble P (sP), respectively: low P (LP) = 6.8 g/kg tP, 3.5 g/kg sP, regular P (RP) = 10.0 g/kg tP, 5.6 g/kg sP, and high P (HP) = 13.0 g/kg tP, 9.3 g/kg sP. Animals were analysed for plasma and bone mineral content, vertebral column deformities (x-ray), vertebral centra stiffness, bone mineralisation pattern and vertebral body microanatomy (cells and connective tissue structures). A low (background) level of deformities was observed on a gross morphological level but no increase and no specific type of vertebral column deformity was associated with either of the three groups. While feed intake was comparable among all diet groups animals fed LP showed a 50% reduction in total calcium (Ca) and P content in abdominal vertebrae and opercula. Regular P and HP animals showed similar levels of total Ca and P in abdominal vertebrae and opercula. Animals in all diet groups showed well-developed vertebral bodies. Low P animals had vertebral centra, neural and haemal arches with large areas of non-mineralised bone. Vertebral centra stiffness in LP animals was reduced accordingly. Regular P and HP animals showed comparable values for vertebral centra stiffness. Non-mineralised vertebral body end plates of LP animals developed a slight inward bending and intervertebral ligaments increased in length and thickness. The cellular and extracellular components of the intervertebral joints remained intact without structural alterations that would indicate the development of vertebral centra compression or fusion. Animals from all three diet groups showed active osteoblasts at the vertebral body growth zone. Despite the three-fold decline in plasma inorganic P in LP animals growth continued at the same rate as in RP and HP animals. It is discussed whether the use of a P-reduced diet under a continuous feeding regime can maintain growth without adverse effects for animal health and welfare. This study further discusses that a HP diet relative to an RP diet has no beneficial effect concerning bone formation, bone mineralisation, growth and prevention of vertebral centra deformities in Atlantic salmon parr.publishedVersio

Phosphorus nutrition in farmed Atlantic salmon (Salmo salar): life stage and temperature effects on bone pathologies

Bone health is important for a viable and ethically sound Atlantic salmon aquaculture industry. Two important risk factors for vertebral deformities are dietary phosphorus and water temperature. Here, we explore the interplay between these two factors during a full production of Atlantic salmon. Salmon were fed one of three diets (low 4.4–5.0 g kg−1, medium 7.1–7.6 g kg−1, or high 9.0–9.7 g kg−1 soluble phosphorus) from 3 to 500 g body weight, followed by a common diet of 7.3 g kg−1 soluble phosphorus until harvest size at 4 kg. Additional groups were included to investigate the effects of water temperatures of 10 vs 16 °C (low and high diets only) and the switching of dietary phosphorus levels (from low to medium or high, from medium to low or high, from high to low or medium), starting at seawater transfer (~100 g body weight) and lasting for 4 months (~500 g body weight). During the experimental feeding period, the low phosphorus diet caused reduced bone mineralization and stiffness and a greater prevalence of vertebral deformities, compared to the medium and high phosphorus diets. However, the prevalence of severely deformed fish at harvest was reduced by switching from the low to either the medium or high phosphorus diets for 4 months after seawater transfer, followed by rearing on the standard commercial feed. Concurrently, switching from either the medium or high to a low phosphorus diet for the same period following seawater transfer had no effect on vertebral deformities at harvest. The higher water temperature for 4 months following seawater transfer increased the severity of deformities at harvest, irrespective of dietary phosphorus. Finally, low dietary phosphorus was associated with increased fillet damage, due to ectopic connective tissue around the spine, at harvest. In conclusion, dietary phosphorus levels of 5 g kg−1 for the initial 4 months in seawater are more of a risk factor for vertebral pathologies if preceded by low, but not medium or high, dietary phosphorus in freshwater. However, dietary phosphorus levels may not play a role in temperature induced radiologically detectable vertebral pathologies. Under the reported growing conditions and diet compositions, a combination of 7.5–7.6 g kg−1 soluble phosphorus during freshwater and 5.0 g kg−1

Zebrafish Tric-b is required for skeletal development and bone cells differentiation

IntroductionTrimeric intracellular potassium channels TRIC-A and -B are endoplasmic reticulum (ER) integral membrane proteins, involved in the regulation of calcium release mediated by ryanodine (RyRs) and inositol 1,4,5-trisphosphate (IP3Rs) receptors, respectively. While TRIC-A is mainly expressed in excitable cells, TRIC-B is ubiquitously distributed at moderate level. TRIC-B deficiency causes a dysregulation of calcium flux from the ER, which impacts on multiple collagen specific chaperones and modifying enzymatic activity, leading to a rare form of osteogenesis imperfecta (OI Type XIV). The relevance of TRIC-B on cell homeostasis and the molecular mechanism behind the disease are still unknown.ResultsIn this study, we exploited zebrafish to elucidate the role of TRIC-B in skeletal tissue. We demonstrated, for the first time, that tmem38a and tmem38b genes encoding Tric-a and -b, respectively are expressed at early developmental stages in zebrafish, but only the latter has a maternal expression. Two zebrafish mutants for tmem38b were generated by CRISPR/Cas9, one carrying an out of frame mutation introducing a premature stop codon (tmem38b-/-) and one with an in frame deletion that removes the highly conserved KEV domain (tmem38bΔ120-7/Δ120-7). In both models collagen type I is under-modified and partially intracellularly retained in the endoplasmic reticulum, as described in individuals affected by OI type XIV. Tmem38b-/- showed a mild skeletal phenotype at the late larval and juvenile stages of development whereas tmem38bΔ120-7/Δ120-7 bone outcome was limited to a reduced vertebral length at 21 dpf. A caudal fin regeneration study pointed towards impaired activity of osteoblasts and osteoclasts associated with mineralization impairment.DiscussionOur data support the requirement of Tric-b during early development and for bone cell differentiation