Ontogeny and functional histochemistry of the digestive and visual systems and other organs during the larval development of the thick-lipped grey mullet, Chelon labrosus

  • Sarasquete C
  • Ortiz-Delgado J
  • Martos-Sitcha J
  • et al.
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Abstract

The main ontogenetic and functional changes of the digestive and visual systems of Chelon labrosus during the first three months from fertilization until 99 days post-hatch (dph) are described. An extended slow growth pattern (3.93%) during the first three weeks of larval life was discernible. A long transitional period of mixed endo-exogenous feeding was noticeable until 13 dph. The majority of organ systems and structures (eye, digestive system, liver, pancreas, thyroid and others) were developed and functional before first feeding (mouth opening at 4 dph). First differentiation of gastric glands was detected at 16 dph, with the appearance of a noticeable pyloric muscular portion or typical gizzard and first developed pyloric caeca from 23 dph. The presence of pinocytotic supranuclear vesicles (5 dph) was still visible in the posterior intestine until the third month of life. The eyes exhibited early differentiation and functionality (pigmentation, first cone-type photoreceptors) during the lecithotrophic phase (2 dph), and development of all typical retinal layers containing different retinal cell types was progressively discernible from 16 dph onwards. From this time, rod photoreceptor precursor cells were incorporated into the basal region of the outer nuclear layer, and increases of two types of photoreceptors (cones and rods) were detected from 23 dph.

Figures

  • Fig. 1. – Presence and utilization of oil globules in eggs (A to C) and larvae (D) of the Chelon labrosus. A, 24 hours after fertilization (haf); B, 32 haf; C, 48 haf; D, 2 hours post-hatching (hph). Scale bars 1.0 mm.
  • Fig. 2. – Oil globule and yolk sac reabsorption in Chelon labrosus larvae. Values are represented as mean ±S.E.M (n=25-30). Thin lines represent exponential regression plots. The equations were estimated taking into account individual larvae. Bar represents the time of mouth opening (4 days post-hatching, dph).
  • Table 1. – Schematic synthesis of the main ontogenetic events occurring in Chelon labrosus larvae during the lecitotrophic (Stage 1) and endoexogenous (Stage 2) phases. Numbers in brackets indicate the age in days post-hatching (dph) Ai, anterior intestine; Bc, blood cells; Bphc, bucopharyngeal cavity; CC, chloride cells; Ep, exocrine pancreas; GaG, gas gland; Hc, hepatocytes; I, intestine; Li, liver; Mc, mucous cells; Md, mandibular; Oe, oesophagus; OG, oil globule; P, periblast; Ph, pharyngeal; Pi, posterior intestine; RtM, rete mirabile; Si, supranuclear inclusions; St, stomach; TB, taste buds; YS, yolk sac; YSL, yolk syncythial layer; Zg, zymogen granules.
  • Table 2. – Schematic synthesis of the main ontogenetic events occurring in Chelon labrosus larvae during exogenous (Stage 3) and metamorphic (Stage 4) phases. Numbers in brackets indicate the age (dph) Bphc, bucopharyngeal cavity; Ca, cardias; Ep, exocrine pancreas; Fu, fundus; GG, gastric glands; ILM, inner limiting membrane; Li, liver; Mc, mucous cells; Oe, oesophagus; OLM, outer limiting membrane; ONL, outer nuclear layer; Pi, posterior intestine; Py, pyloric region; Rc, rod cells; Si, supranuclear inclusions; St, stomach.
  • Table 3. – Equation and coefficient of determination (R2) of each regression period for mean dry weight and mean total length values (see Figs 3 and 4). DW, dry weight; T, age (dph); TL, total length.
  • Fig. 3. – Mean weight values of growth in larvae and juveniles of Chelon labrosus as a function of age (1 to 99 dph). Data are plotted on a logarithmic ordinate grid and values are represented as mean ±S.E.M (n=10-30). Linear regression equations for each growth stage (bold lines) were estimated by the least-squares regression method using log-transformed data. Vertical dashed lines indicate inflexion periods (i.e. change in regression). Bar represents the moment of the mouth opening (4 days post-hatching, dph). * Represents statistically significant differences between the slope of periods A and B (t=6.532, P<0.05) and periods B and C (t=6.398, P<0.05).
  • Fig. 4. – A, mean total length data of growth in larvae and juveniles of Chelon labrosus as a function of age from 1 to 99 dph. Data are plotted on a logarithmic ordinate grid, and values are represented as mean ±S.E.M (n=10-30). Linear regression equations for each distinct growth stage (bold lines) were estimated by the least-squares regression method using log-transformed data. Bar represents the moment of mouth opening (4 dph).
  • Fig. 5. – Histological sections of the Chelon labrosus during the endo-exotrophic stages. A, 0 days post-hatching (dph) larvae showing an acidophilic homogeneous yolk matrix and adhered lipid droplet close to a straight digestive duct. A tubular heart anlage is seen within pericardiac cavity. B, larvae at 2-3 dph showing a yolk mass with a reduced volume. Note mouth opened. Incipient gills are also discernible at this stage. C, progressive differentiation of the digestive system into different segments (buccopharynx, oesophagus, stomach anlage, anterior and posterior intestine). D, larvae at 3-4 dph showing a slight yolk sac and a still-present oil globule. Exocrine with acidophilic zymogen granules and endocrine (Langerhans islets) pancreatic portions are visible. Swim bladder anlage is also detected at this stage. E, periblast enclosing residual yolk and oil globule is visible in larvae by 7 dph. F and G, larvae at 4-5 dph showing the bucopahryngeal cavity (F) with pharyngeal teeth and a developing oesophagus with a pseudo-stratified epithelium and first mucous cells and an incipient stomach (G) as a small pocket covered by a thickened mucosal layer which lacks gastric glands. Renal tubules are proliferating at this stage. H, first mucous cells detected in buccopharyngeal cavity and oesophagus at 5 dph. Thickening of mucous and muscular layer are also detected at this stage. I, a more developed and folded stomach with a thickening of mucosal layer is also seen at 5 dph. J, 5-6 dph larvae showing pyloric sphincter separating the anterior intestinal portion from the developing anlage stomach. Noticeable increase in size of liver is also seen at this stage. K, first mucous cells present on anterior intestine at 5-6 dph. Note the presence of ileorectal valve separating anterior intestine from the bending posterior intestinal portion. Exocrine pancreas increased in size at this stage. L, acidophilic supranuclear inclusions first detected in the posterior intestine at 6 dph. M to O, larvae at 8-9 dph showing osmiophilic lipid inclusions in enterocytes from the anterior and median intestine and within cytoplasm of hepatocytes. Scale bar represents (A, B) 500 μm; (C-E) 300 μm; (F-O) 200 μm. Haematoxylin-VOF (A-E, H, I, L). Haematoxylin-eosin (F, G, J, K, M). Osmium tetroxide posfixation (N, O). AI, anterior intestine; BpC, bucopharyngeal cavity; DT, digestive tube; ExP, exocrine pancreas; G, gut; GA, gills anlage; IRV, ileorectal valve; L, liver; LI, Langerhans islet; Lin, lipid inclusions; MC, mucous cells; Oe, oesophagus; OG, oil globule; P, periblast; PC, pericardic cavity; PI, posterior intestine; PS, pyloric sphincter; PT, pharyngeal teeth; RT, renal tubules; SB, swim bladder; SNI, supranuclear inclusions; Sp, spleen; St, stomach; YS, yolk sac; ZG, zymogen granules.

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Sarasquete, C., Ortiz-Delgado, J. B., Martos-Sitcha, J. A., De las Heras, V., Yúfera, M., & Martínez-Rodríguez, G. (2014). Ontogeny and functional histochemistry of the digestive and visual systems and other organs during the larval development of the thick-lipped grey mullet, Chelon labrosus. Scientia Marina, 78(4), 473–491. https://doi.org/10.3989/scimar.04091.27b

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