Sperm Competition and Mode of Fertilization in the Grass Goby Zosterisessor Ophiocephalus (Teleostei: Gobiidae) 1 1 2 M

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Sperm Competition and Mode of Fertilization in the Grass Goby Zosterisessor Ophiocephalus (Teleostei: Gobiidae) 1 1 2 M JOURNAL OF EXPERIMENTAL ZOOLOGY 283:81–90 (1999) Sperm Competition and Mode of Fertilization in the Grass Goby Zosterisessor ophiocephalus (Teleostei: Gobiidae) 1 1 2 M. SCAGGIANTE, C. MAZZOLDI, C.W. PETERSEN, AND M.B. RASOTTO1* 1Dipartimento di Biologia, Università di Padova, 35121 Padova, Italy 2College of the Atlantic, Bar Harbor, Maine 04609 ABSTRACT Accessory structures to the male reproductive system are known in several fami- lies of teleost, but their role in sperm production patterns and fertilization dynamics is still un- clear. The intraspecific variability in seminal vesicles, shown by the grass goby Zosterisessor ophiocephalus, presents an opportunity to examine both its possible correlation to alternative male mating tactics and the function of these accessory structures. In this species, males are known to release sperm in the form of sperm trails, bands of mucosubstances in which sperm are embedded. Surgical removal of seminal vesicles and histochemistry demonstrate that mucins, involved in the production of sperm trails, are secreted by the seminal vesicles. Gametes show a high longev- ity; sperm motility lasts on average 80 minutes. Eggs can be fertilized for several hours and do not need to be laid over trails because sperm are able to reach them via the surrounding water. Gonosomatic and seminal vesicle somatic indices, histology and histochemistry of gonads and seminal vesicles, sperm counts, and sperm trail longevity, suggest the presence of alternative male mating tactics in this species. Larger males have smaller testes and larger seminal vesicles compared to those of smaller males. The major role of seminal vesicles is mucin secretion in larger males, sperm storage in smaller ones. Trails of larger males last longer and release sperm more constantly over time than those of smaller males. Overall differences between males support the presence of differences in the intensity of sperm competition, with larger males performing nesting behaviour while smaller ones sneak spawnings. J. Exp. Zool. 283:81 90, 1999. © 1999 Wiley-Liss, Inc. Sperm competition, defined as the competition them. It is unclear how these structures are between the sperm of two or more males to fertil- related to sperm production patterns, both evolu- ize the eggs of a female (Parker, ’70), is well known tionarily and functionally. Current sperm compe- to strongly influence not only the behaviour of tition models that use gonosomatic indices to males but also their reproductive anatomy and estimate sperm production within or between spe- physiology (Smith, ’84). In fish, it has been dem- cies assume that functions other than sperm pro- onstrated that males exposed to higher levels of duction are either not important or are constant sperm competition have relatively larger testes across comparisons. and release an absolute higher number of sperm The family Gobiidae is a large family of small, per spawn or ejaculate (Robertson and Warner, freshwater and marine species with demersal ’78; Warner and Robertson, ’78; Shapiro et al., ’94; spawning and male parental care. Male sperm Stockley et al., ’97; Petersen and Warner, ’98). duct glands, usually called seminal vesicles, are Testes and the associated structures of the male accessory structures of the reproductive appara- reproductive system have roles that go beyond tus of all species of the suborder Gobioidea (Miller, sperm production. The production of substances ’92). However, neither the significance of their that may serve hormonal or pheromonal roles, fa- cilitating sperm delivery and sperm storage, are important roles of the testis and its accessory Grant sponsors: Italian Ministero dell’Università e della Ricerca structures (Fishelson, ’91; Van den Hurk and Scientifica e Tecnologica; U.S. National Science Foundation. *Correspondence to: M.B. Rasotto, Dip. Biologia, Viale G. Colombo Resink, ’92). These accessory structures often are 3, 35121 Padova, Italy. E-mail: [email protected] embedded within the testes, or lay adjacent to Received 1 December 1997; Accepted 28 April 1998 © 1999 WILEY-LISS, INC. 82 M. SCAGGIANTE ET AL. high variability in the development and secretory seminal vesicles varies intraspecifically are not activity, both across and within species (Miller, known. The second question regards the evolu- ’84; Fishelson, ’91; Joyeux et al., ’91), nor their tionary significance of sperm trail production and roles in reproduction, are well understood. Semi- the asynchronous gamete release between males nal vesicles are known to secrete mucosubstances and females. If sperm trails or similar sperm re- in many gobiid species (Miller, ’84; Lahnsteiner lease mechanisms are a function of accessory et al., ’90; Fishelson, ’91). In several other teleost structures, then by studying intraspecific and in- families of demersal spawners with male pa- terspecific variation in its development we may rental care, such as Blenniidae, Labrisomidae, gain insight into the selective forces that shaped Tripterygiidae, Cottidae, Gasterosteidae, Heter- the evolution of these structures and their prod- opneustidae, etc., males also release mucins ucts. Marconato et al. (’96) suggested one pos- produced either by structures accessory to the sible advantage of sperm trails: by releasing gonads (Lahnsteiner et al., ’90; Rasotto, ’95) or sperm in trails, territorial males could defend by an hypertrophied kidney (Bucher and Hofer, territories while active sperm were still being ’93). One hypothesized role of these mucosub- released in a nest. stances is the production of sperm trails: bands The specific aims of this study were (1) to evalu- of viscous materials in which sperm are embed- ate the investment in both gonads and seminal ded when released (Marconato et al., ’96; Ota vesicles in males of different size; (2) to clarify et al., ’96). Many fishes, including some with the function of seminal vesicles; (3) to compare these accessory structures, are known to have sperm trail longevity and sperm viability of males alternative male mating tactics that often re- of different size; and (4) to study fertilization sult in different levels of sperm competition and dynamics. mating success for different individuals in a population (Taborsky, ’94). Intraspecific vari- MATERIALS AND METHODS ability in seminal vesicles has been described The grass goby, Zosterisessor ophiocephalus in some gobiid species, but its possible correla- (Pallas, 1822), is a large coastal gobiid inhabiting tion with different male mating opportunities the seagrass meadows of Zostera (Z. marina and or different reproductive tactics has never been Z. nolti) in shallow brackish water. During the investigated. reproductive season, lasting from March to May, To examine the hypotheses of seminal vesicles territorial males dig a burrow under the seagrass function(s) and their possible relationship to male rhizomes. Females lay the eggs on the ceiling of mating tactics, we studied the Mediterranean the burrow, attaching them to overhanging Zostera grass goby, Zosterisessor ophiocephalus. The spe- rhizomes. The eggs diameters are 0.88–1.21 mm, cies has seminal vesicles known to produce mu- and hatching occurs after 10 days at a water tem- cins (Lahnsteiner et al., ’90) and presents a perature of 15°C. Males defend the nest and take conspicuous variability in the size of mature males care of the eggs laid by one to several females, (De Girolamo, ’94). A recent study demonstrated cleaning and fanning them until hatching (Gan- that in Z. ophiocephalus and two other goby spe- dolfi et al., ’91; De Girolamo, ’94). In the Venetian cies, sperm trails are released by males before fe- Lagoon (Northern Adriatic Sea, Italy), the grass males start to lay eggs, and the trails continue to goby reaches sexual maturity after one year and release active sperm over prolonged periods of lives for up to two and a half years, reaching a time (Marconato et al., ’96). In these species, maximum total length of 25 cm (De Girolamo, ’94). sperm are released directly onto the nest surface. Mature males and females of Z. ophiocephalus This asynchronous mechanism of gamete release were collected during the reproductive seasons is atypical for teleosts, where successful external 1995 and 1996, in the Venetian Lagoon using per- fertilization is assumed to require the synchro- manent fish traps. Males and females were rec- nous release of gametes by both partners (Breder ognized by the sexually dimorphic genital papillae and Rosen, ’66; Jamieson, ’91). (Gandolfi et al., ’91) and were kept separate in Two questions arise from these observations in large stock tanks. Each aquarium was provided Z. ophiocephalus. The first concerns the glandu- with a sandy bottom and with artificial shelters. lar production of sperm trails. It has been sug- Seawater was renewed daily, and temperature gested that seminal vesicles play a major role in range was kept at 17–18°C. Light regime followed sperm trail production, but whether this is their natural conditions, and fishes were fed once a day primary function and whether the function of with fresh black mussel meat. SPERM COMPETITION AND FERTILIZATION IN A GOBY 83 Gonosomatic and seminal vesicle sperm trail, laid on a clean slide, was stained by somatic indices PAS; then the males were deeply anaesthetized in an aerated seawater solution of MS222 for 6–8 In 1995 and 1996, respectively, 129 (size range: min. A 1.5 cm ventral incision was made close to 8.5–22.0 cm total length) and 84 (size range: 8.1– the genital papilla, the seminal vesicles were re- 22.6 cm total length) freshly caught males were moved, and the incision was closed with 2.0 sur- sampled. If not already dead, they were sacrificed gical silk. The gills periodically were irrigated with with an excess of anaesthetic (MS222). Total fresh seawater during surgery. Males recovered length (TL) and weight of body, gonads and semi- rapidly, were immediately returned to their home nal vesicles of every specimen were measured (to tanks, and were fed 12 hr later. Following six days the nearest mm and 0.01 g).
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