Early stages of test formation in larva of Ascidia malaca (Tunicata, Ascidiacea): ultrastructural and cytochemical investigations.

The swimming larvae of ascidians are entirely covered by a hyalin coat called tunic, or test. This covering consists of two cuticular layers, C1 and C2, which surround an inner compartment composed of an amorphous hyalin matrix with numerous fibrils dispersed inside. Data from the literature agree on the key role played by the cells of the larval ectodermic layer in the synthesis and secretion of larval test components. In the present article are reported ultrastructural and cytochemical investigations made during test formation in the swimming larva of Ascidia malaca. Besides confirming the role played by ectodermic cells during the early stages of test formation, the investigations highlight the way in which the fibrillar component of the test is synthetized and secreted. At the ultrastructural level it has been evidenced that the C1 and C2 cuticular layers originate from the tight packing of fibrils. Based on the data reported in the present study, it is hypothesized that while a relevant part of the fibrils, once secreted, remains dispersed inside the matrix of the inner compartment of the test, quite likely in order to increase its consistency, packing of the remaining fibrils leads to the formation of the C1 and C2 cuticular layers. Packing of the fibrils in C1 and C2 could be favoured by their chemically adhesive nature. This hypothesis is strongly supported by the herewith reported results of the cytochemical investigations carried out on the test of the swimming larva of A. malaca. The cytochemical PA-TCH-SP reaction has in fact evidenced that both fibril types, i.e. those dispersed inside the inner compartment and those packed in the C1 and C2 cuticular layers, are constituted by glycoproteins and/or proteoglycans substances whose adhesive properties are well documented in the literature.