ALBERT

Notes: Several polyacrylamide gel electrophoresis techniques were used to study developmental changes in myofibrillar protein composition and parvalbumin distribution in the myotomal muscle of Brycon moorei. Two myosin LC2 chains and two troponin I isoforms were successively detected. Up to four troponin T isoforms were synthesized. Slow red-muscle myofibrils from adult fish showed no common component (except actin) with larval, juvenile or adult fast white-muscle myofibrils. During growth of B. moorei, two classes of parvalbumin isoforms were sequentially expressed: larval PA I, PA IIa, and PA IIb and adult PA III. In adult fish, the content in Tn T-2 isoform decreased from the anterior to the posterior myomeres, in favour of Tn T-1 and Tn T-4. The parvalbumin content also diminished from the rostral to the caudal muscle. The fast rate of transition from larval to adult isoforms appeared to parallel the extremely fast growth of B. moorei. Sequential expression of these isoforms presumably reflected variations in the contractile properties of the muscle fibres, required by changes in physiological demands of the propulsive musculature.

Notes: The inception and development of the cartilaginous cephalis skeleton of Chrysichthys auratus is described from hatching to about 18 days post-hatching. At hatching, no skeletal structure is present. Not until day 3 do clearly delimited cranial primordia become apparent. As in many siluriforms, the neurocranium is platybasic from the start, the suspensorium constitutes, with Meckel's cartilage and the hyoid bar, a single cartilaginous element, and the junction between the front and rear of the neurocranium is complete on day 4. By day 8 the quadratomandibular joint has formed and the tectum posterius has appeared. Cartilage reduction first affects the trabecular bars, then, markedly, the visceral arches. By day 18 the braincase floor has almost disappeared.

Notes: Parvalbumin isotypes PA II, PA III, PA IVa, and PA IVb were isolated by chromatography from trunk white muscle of barbel and physicochemically characterized. Electrospray ionization mass spectroscopy revealed that PA II has a lower molecular weight than the other isotypes and that PA IVa and PA IVb each consist of two subforms. Isotype distribution was studied by polyacrylamide gel electrophoresis. In adult fish, the total parvalbumin titre decreased and the isotype distribution varied from the anterior to the posterior myotomes. In the course of barbel development, the total parvalbumin titre increased rapidly as fish standard length increased from 1·3 to 5 cm; then sloped down gently as the length increased to 60 cm. At least six parvalbumin isotypes were identified, three of which are different forms (a, b, and c) of PA II. These three forms were present together at the larval stage, but PA IIc and chiefly PA IIb appeared as early isotypes, contrary to PA IIa which was present until the adult period. Later PA IVb accounted for up to 90% of the total parvalbumin content; PA III and PA IVa are minor adult isotypes. Temporal and spatial variations in the total parvalbumin titre and in the differential expression of barbel parvalbumin isotypes very likely reflected the functional requirements of the fish axial musculature according to fish size and myotome location. Physiologically, the larval isotypes could promote faster relaxation of fast fibres than the adult isotypes, and hence favour shorter contraction times.

Notes: At hatching, Heterobranchus longifilis does not display any primordia of the cephalic skeleton. The latter appears 12 h post–hatching and develops in three stages up to day 16. The first stage (12 h to 2 days) involves almost exclusively the development of the chondrocranium. During the second period (days 3–8), dermal elements of the splanchnocranium appear. The final stage is marked by resorption of the cartilages, progressively replaced by ossifications (days 10–16). At their appearance the elements of the splanchnocranium are fused together, as are the first neurocranial elements. Later, the splanchnocranium splits up. By the time the yolk sac is completely resorbed, the buccal and pharyngeal jaws are present, the suspensoria and hyoid bars are partially developed, and the parasphenoid partially closes the hypophyseal fenestra. These structures delimit a buccal cavity that is probably functional, i.e. capable of participating in the intake of exogenous food. Next to continue its development is principally the splanchnocranium, completing the walls of the buccal cavity. Cartilage resorption parallels the appearance of endochondral ossifications (except for the trabecular bars). Braincase closure begins to accelerate once the buccal system is complete.

Notes: The inception, and development of the cephalic skeleton of Barbus barbus from hatching to 24 days passes through periods of fast and slow growth; these rates are not the same in different parts of the skull. Trabeculae, parachordal plates, Meckelian cartilages and hyposymplectics are present at hatching. Then the cartilaginous floor of the neurocranium develops, the pars quadrata, the hyoid bars and branchial arches elements appear shortly before the first movable dermal bones, the dentaries, maxillae and opercles. The first bone of the braincase to appear is the parasphenoid; other bones develop subsequently and at the same time: the angular, quadrate, interopercle and fifth ceratobranchial. Later the splanchnocranium continues to develop at a relatively fast rate while the neurocranium shows little growth. The braincase does not begin to close before the 24th day, nor do the first bones of the skull roof appear, while the bucco-pharyngeal apparatus is complete, having the adult shape. The early constitution of the latter structures seems to be linked with the mechanical demands of biological functions such as breathing and feeding.

Notes: At hatching Scophthalmus maximus shows no cartilaginous and no bony structure. Meckeľs cartilages appear when the fry are 1 day old, followed on day 2, by formation of the trabecular bars, fused at the outset to form a trabecula communis. Concurrently, the palatoquadrates complete the mandibular arch, and the first two pairs of ceratobranchials, associated with a pair of hyoid bars, form the beginnings of the hyobranchial system. By day 3, the parachordals have fused with the trabecular bars, the hyosymplectics have linked to the hyoid bars by interhyals, and the first four pairs of ceratobranchials have appeared. The first bony structures appear: the preoperculars. On day 8, the frontals develop above the orbits and the maxillaries and dentaries appear. On day 10, the primordia of the taeniae marginales appear, the palatoquadrates bear a pterygoid process, and to the branchial basket have been added the fifth pair of ceratobranchials and the four pairs of epibranchials. On day 12, both pairs of posterior pharyngobranchials are present. The premaxillaries develop in front of the maxillaires, and retroarticulars and the angulars complete the lower jaws. On day 13, a thin parasphenoid contributes to the floor of the neurocranium, and ectopterygoids and entopterygoids to the splanchnocranium. The set of opercular bones is complete. On day 15, the tectum synoticum closes the braincase posteriorly. The splanchnocranium possesses a basihyal and the pharyngobranchials of the first epibranchials. On day 18, the tectum posterius completes the dome of the braincase. The rear end and lateral walls of the skull are formed by the basioccipital, the exoccipitals, the pterotics, and the parietals. The suspensorium is nearly complete. From day 10, the first resorptions begin in parallel with the construction of the chondrocranium. Meckeľs cartilages each split in two, then the posterior part of the trabecular bars disappears. On day 23, the right taenia marginalis separates from the lamina orbitonasalis and curves towards the centre. Simultaneously, the right eye begins its migration to the left. This is the only metamorphosis-linked asymmetry to appear during the development of the chondrocranium. On day 25, many more bony structures appear, a characteristic of this stage: the nasals, lateral ethmoids, mesethmoid, sphenotics, prootics, pleurosphenoids, epiotics, and supraoccipital. From this stage on, the bony structures continue to develop, while the front of the neurocranium and the jaws undergo a deep remodelling due to metamorphosis. The left taenia marginalis does not appear reduced until day 29. By day 45, there remain only a few small elements of the cartilaginous skull.

Notes: Muscle proteins were investigated in two large European barbels, Barbus barbus and B. meridionalis, and in four small tropical barbels native to SE Asia: B. conchonius, B. tetrazona, B. sachsi and B. titteya. Polyacrylamide gel electrophoresis was used to analyse myosin heavy and light chains and parvalbumin isotypes from white trunk muscle. Each species could be biochemically identified. The myosin subunit and parvalbumin isotype patterns obtained for the two European barbels were similar. The Asian barbels, on the other hand, not only differed from the European species but displayed a greater diversity within their group. These biochemical results are largely in agreement with morphological and genetic data, but fail to substantiate suggested close relationships between Asian barbel species.

Notes: The white-muscle parvalbumin isoforms of Clarias gariepinus, Heterobranchus longifilis and Chrysichthys auratus were purified and their physicochemical parameters determined. The three catfish isoforms are distinct but those of C. gariepinus and H. longifilis are more similar. In the course of development, the successive appearance of larval and adult parvalbumins was observed. Larval isoforms (PA I, PA IIa, PA IIb) displayed a lower isoelectric point (pI) and molecular mass than adult ones (PA IIc, PA IIIa, PA IIIb, PA III, PA IV). The PA IIa isoform appeared as an omnipresent typical larval isoform. PA IIb appeared mostly larval, being insignificant in adult specimens; its physicochemical features were the same in the three catfish species. In Chrysichthys auratus, there were three PA II isoforms, one appearing as an adult isoform (PA IIc). The fact that the two types of parvalbumin isoforms appear at different times should reflect specific physiological needs (mobility, feeding) of different developmental stages.