The ovary of the bignose fanskate Sympterygia acuta Garman, 1877 (Chondrichthyes, Rajidae) in the Bahía Blanca estuary, Argentina: morphology and reproductive features Díaz-Andrade, MC., Galíndez, E.* and Estecondo, S. Laboratorio de Histología Animal, Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur – UNS, San Juan 670, 8000, Bahía Blanca, Argentina *e-mail: galí[email protected] Received July 31, 2007 – Accepted September 10, 2007 – Distributed May 31, 2009 (With 13 figures) Abstract Chondrichthyes have become an important economic resource in recent years. In spite of this importance, there is little knowledge about their reproductive biology, especially of species from the south-western Atlantic. In this work, we study the morphology and histology of the ovary of Sympterygia acuta. The results show that oogonia were present only in the immature females (stage I of maturity scale), whereas specimens in stages II and III of maturity displayed oocytes in all developmental steps. The most important histological features that vary throughout oocyte development are the presence or absence of yolk, the number of types and layers of follicular cells and the degree of development of the thecae. Follicular cells are, at least, of two different types. Finally, a new point of view for the determination of maturity stages in the field based on the size of follicles is discussed. Keywords: Chondrichthyes, folliculogenesis, maturity stage, reproductive biology, Sympterygia acuta. O ovário da bignose fanskate Sympterygia acuta Garman, 1877 (Chondrichthyes, Rajidae) no estuário de Bahía Blanca, Argentina: morfologia e características reprodutivas Resumo Os Chondrichthyes tornaram-se num importante recurso econômico nos últimos anos. Apesar da sua importância, é muito pouco o que se conhece da sua biologia reprodutiva, especialmente das espécies do Atlântico sul-ocidental. Neste trabalho, foi estudada a morfologia e histologia do ovário de Sympterygia acuta. Os resultados mostraram que as oogonias somente encontraram-se presentes nas fêmeas imaturas (fase I de escala de maturidade) e que os animais em estágio II e III de maturidade exibiram oocitos em todas as etapas de desenvolvimento. As características histológicas mais importantes que variam ao longo de desenvolvimento dos oocitos são a presença da suarda, o tipo e quantidade de celas foliculares e o grau do desenvolvimento do thecae. Existem pelo menos dois tipos de células foliculares. Finalmente se discute um novo ponto de vista para a determinação dos estágios de maturidade no campo, baseado no tamanho dos folículos. Palavras-chave: Chondrichthyes, folliculogenesis, fase de maturidade, biologia reprodutiva, Sympterygia acuta. 1. Introduction Chondrichthyes appeared on earth 400 million years diversity in reproductive strategies, the group is charac- ago. They constitute a diverse group in all aspects of terized by low growth, an extensive period of gestation lifestyle, particularly in their mode of reproduction and a scarce number of large offspring, resulting in low (McEachran and Dunn, 1998). All modern chondrichthy- reproductive potential (Hoenig and Gruber, 1990). ans exhibit internal fertilization and specialized genital At least half of the Chondrichthyes belong to the or- tracts adapted for uterine gestation (Tricas, 1980). They der Rajiformes, being the Rajidae family the most im- are either oviparous or viviparous with four modes of portant. All skates are oviparous producing large oocytes viviparity based on the source of nutrition for the devel- with considerable yolk supplies. They encapsulate the oping embryo (Koob and Callard, 1991). In spite of this eggs in complex egg cases produced by the oviducal Braz. J. Biol., 69(2): 405-413, 2009 405 Díaz-Andrade, MC., Galíndez, E. and Estecondo, S. gland and lay them in muddy or sandy areas where they For each specimen, at least one ovary was cut in small adhere to small pieces of rock, shell or seaweed. This egg pieces for histology and in six specimens one ovary was case constitutes the only protection against pathogens or dissected in toto for follicular countdown. This mate- predators (Kormanik, 1993). rial and small pieces of the liver were fixed in Bouin’s Sympterygia acuta is an endemic species of the fixative or formaldehyde 10% in seawater, dehydrated Argentinean Province with a latitudinal distribution be- through a graded series of alcohols and embedded in tween 22° and 43° 20’ S (De-Queiroz, 1995). It can be paraffin wax. Then, 5-10μ m-thick sections were stained found from coastal waters to 188 m of depth (Meneses by Masson’s trichromic stain, haematoxylin-eosin, al- and Paesch, 2003), although it is caught mainly at 50 m cian blue technique at 2.5 pH level, periodic acid Schiff of depth (Massa et al., 2004). The Bahía Blanca estu- reaction, Gomori technique for reticulum and orceine. ary is known for its use by fishes to mate and give birth Selected sections were photographed using an Olympus (López-Cazorla, 1987). S. acuta is present in this estuary BX51 light microscope equipped with an Olympus throughout the year, with an abundance peak from the C-7070 digital camera. middle spring to summer (Mabragaña et al., 2002) con- The ovaries fixed in toto were carefully opened and stituting an important economic resource (Massa et al., each oocyte inside was measured in millimetres using a 2003). The UICN Red List for threatened species (2006) SOMET calibre with a 1/20 mm precision. The data ob- has included this species since 2004 considering it “vul- tained was grouped at intervals of increasing amplitude nerable” (A2bd). Nevertheless, S. acuta, along with to compensate the asymmetry of the variate. other skates and demersal sharks, is a common by-catch A classification limit was identified macroscopically in the coastal and continental shelf bottom-trawl fisher- at 3.5 mm corresponding to the smaller size at which ies in the Argentine-Uruguayan Common Fishing Zone oocytes were yellow prior to the fixation. On the other (Meneses and Paesch, 2003). hand, another limit was identified microscopically at In spite of the evolutionary and economic impor- 1 mm, corresponding to the smaller size at which yolk tance of chondrichthyans, there are a scarce number of was present in the ooplasm. investigations with regards to the ovary structure and they have been conducted mainly in species from in- 3. Results tensive fishing zones (Pratt, 1988, Hamlett et al., 1999, Hamlett and Koob, 1999, Prisco et al., 2001, 2002a,b; Morphometric data of exemplars are shown in Stehmann, 2002, Storrie, 2004, Oddone et al., 2005). Table 1. About the genus Sympterygia there are very few investi- 3.1. Macroscopic description of the ovary gations and they treat mainly ecological features (Pires, Both ovaries were approximately of the same size, 1987, De-Queiroz, 1984, 1995), parasitology (Romera, elongated and slightly compressed dorsoventrally. They 1993, Tanzola et al., 1998) and some reproductive fea- were associated with the epigonal organ that extended tures (Mabragaña et al., 2002, Oddone and Vooren, 2002, beyond the length of the ovary. In a maturing or mature Oddone and Velasco, 2004, Galíndez and Estecondo, ovary (stages II and III of maturation scale) follicles of 2008). various sizes were present at any one time supported by The present work represents a contribution to the loose connective tissue. Even though follicles were inter- knowledge of the structure of the ovary of S. acuta, the features that characterize the different stages of follicu- logenesis and some remarks on the reproductive biology Table 1. Morphometric data of exemplars. of this commercially important and vulnerable species. Maturity Tot. Disc Disc 2. Materials and Methods stage length length width (macroscopic) (cm) (cm) (cm) A total of 12 females of S. acuta were collected Sa 1 III 50.0 28.0 28.0 monthly by line fishing in the inner and medium zones Sa 2 III 54.0 28.0 29.0 of the Bahía Blanca estuary (61° 30’-62° 30’ W and Sa 5 III 54.5 31.0 31.5 38° 45’-39° 30’ S). This number of exemplars was con- sidered appropriate due to the vulnerable status and the Sa 6 III 53.5 28.0 29.0 decline of its biomass in the stocks. The total length and Sa 7 III 50.0 28.0 31.0 disc width and length of each specimen were measured. Sa 8 III 58.0 30.0 33.0 Each animal was humanely terminated by blunt trauma Sa 10 III 56.0 30.0 31.0 to the cranium and then dissected. Once the ventral cav- Sa 11 III 56.6 30.0 31.0 ity was opened, the sexual maturity of the females was Sa 3 II 49.0 32.0 28.5 determined following the Braccini and Chiaramonte Sa 4 II 50.0 32.0 29.0 (2002) criteria. This standard technique, common on board, is more than simple but in agreement, in general Sa 12 II 48.0 27.5 28.0 terms, with that proposed by Stehmann (2002). Sa 9 I 31.0 17.0 22.0 406 Braz. J. Biol., 69(2): 405-413, 2009 The ovary of S. acuta mingled in different degrees of maturation, an increase mately 55 μm, some enlarged globed-shaped cells ap- in size from cranial to caudal and from dorsal to ventral pear between follicular cells. could be recognized externally. At ~180 μm (primary follicles) follicular cells be- come cuboids and arrayed in one layer (Figure 4), be- 3.2. Microscopic description of the ovary tween them there are a great number of the globed-shaped In the immature specimen, the ovary appears as a cells (Figure 5). The zona pellucida is a homogeneous small structure at the medial cranial side of the epigo- hyaline smooth line of 2 μm thick. The oocyte plasmale- nal organ. As ovary development goes on, the epigonal ma folds densely against the zona pellucida and appears organ withdraws and becomes restricted to a small pos- terior outline and the space between oocytes.