Morphology, fine structure, biochemistry, and function of the spermatic ducts in marine fish

The spermatic ducts and the testicular efferent ducts were investigated in different marine teleost fish species (Diplodus sargus, Mullus barbatus, Thalassoma pavo, Trachinus draco, Uranuscopus scaber, Sparisoma cretense, Synodon saurus). From the morphological, histological, fine structural and biochemical investigations it appeared that the testicular main ducts and spermatic ducts of the investigated marine fish have the following functions: storage of spermatozoa, monosacharide synthesis for nutrition of spermatozoa, synthesis of steroid glucuronides, synthesis of seminal plasma proteins, formation of a ionic gradient in the seminal fluid and phagocytotic activity. Species-specific differences were only found in the morphology of the gonads and in the histology of the spermatic duct epithelium.     

Comparative morphology of male reproductive systems in Mediterranean blennies (Blenniidae)

The male reproductive organs of 16 species of Mediterranean Blenniidae ( Aidablennius sphynx, Blennius ocellaris, Coryphoblennius galerita, Lipophrys adriaticus, L. canevae, L. dalmatinus, L. nigriceps, Parablennius gattorugine, P. incognitus, P. sanguinolentus, P. rouxi, P. tentacularis, P. zvonimiri, Paralipophrys trigloides, Salaria pavo and Scartella cristata ) consist of pairs of testes, testicular glands, spermatic ducts, and blind pouches. Three main types of accessory sex organs were found by comparing the external morphology of the male gonads. Differences between species were observed in the volume of the testicular gland in relation to the volume of the testis and in the size and length of the spermatic ducts, and blind pouches. The anatomy of the testicular glands of all species investigated do not differ. Each gland consists of ducts that appear to be tubules which terminate at the testis periphery on one side and at the spermatic duct on the other side. Contrary to previous claims, A. sphynx has no fat body in place of the testicular gland; the gland of this species was not distinguishable from that of the other species investigated. In the Lipophrys species, in P. trigloides , and in B. ocellaris , a transition zone between testis and testicular gland is present. The testicular blind pouches empty into the spermatic ducts, into the ureter, or separately on the genital papilla. In most species, the epithelium has no or low folds, while in S. pavo it possesses high folds that nearly fill the lumen of the blind pouches. The morphological results are discussed in connection with taxonomy, ecology, and behaviour of the fishes.     

Testicular main ducts and spermatic ducts in some cyprinid fishes I. Morphology, fine structure and histochemistry

Alburnus alburnus, Leuciscus cephalus and Vimba vimba efferent duct systems of the male gonads consist of testicular main ducts and spermatic ducts. These have similar histological, fine structural and (enzyme–) histochemical characteristics and function in (1) storage and (2) nutrition of spermatozoa, (3) synthesis of steroid glucuronides, (4) secretion of proteins and enzymes (5) formation of an ionic gradient in the seminal fluid and (6) they have auto– and heterophagocytotic activities. Therefore testicular main ducts and spermatic ducts are important in the formation of the seminal fluid.     

The spermatic ducts of salmonid fishes (Salmonidae, Teleostei). Morphology, histochemistry and composition of the secretion

Using light and electron microscopy, enzyme-histochemistry thin-layer chromatrography and photometry the functions of the spermatic ducts in salmonid fishes (rainbow trout, Oncorhynchus mykiss , Arctic charr, Salvelinus alpinus ; grayling, Thymallus thymallus ; whitefish, Coregonus sp.) were investigated. During spawning the spermatic duct of the Salmonidae has a high secretory activity: it synthesizes steroids, lipids (triglycerides, fatty acids, cholesterol esters, phosphatidylcholine), monosaccharides (glucose, fructose), proteins and enzymes (acid and alkaline phosphatase, glucuronidase and proteases). It is important for storage and resorption of spermatozoa. Interspecific differences exist in the storage capacity for semen.     

Morphology of Male Reproductive Systems in Ephemeroptera: Intrinsic Musculature

Although the Ephemeroptera have been studied over a long period of time, there are still few studies on the morphology of male reproductive system. The spermatic ducts are considered conserved among different Ephemeroptera groups. However, previous studies distinguished different organizational patterns of the spermatic duct intrinsic musculature. This study describes the morphology of the spermatic ducts, especially their musculature, in six species of Ephemeroptera, representing five families. We have observed variations in the organizational pattern of the spermatic ducts, even between species from the same family. Moreover, all species studied had intrinsic musculature in the spermatic ducts although with different organizational patterns. Thus, we believe that this musculature is important to move the spermatozoa along the ducts of all Ephemeroptera and not only of those with aflagellated spermatozoa (Leptophlebiidae). The variations in musculature organization must be related to differences in reproductive physiology (i.e., copula duration) and not only with spermatozoa characteristics.     

Histochemical analysis of sperm storage in Helicolenus dactylopterus dactylopterus (Teleostei: Scorpaenidae).

The bluemouth rockfish, Helicolenus dactylopterus dactylopterus (De la Roche, 1809), is a zygoparous species with internal fertilization. The male urogenital papilla acts as the copulating organ, and the females retain the spermatozoa in their ovaries for up to 10 months. The objective of this study is to extend our knowledge of the mechanisms that allow the sperm to be retained in the ovaries for prolonged periods. To this end, we analyze the histochemical properties of: 1) the epithelium of the testicular sperm duct, 2) the sperm of the males, 3) the internal epithelium of the ovary wall, 4) the ovarian fluid, and 5) the spermatozoa storage crypts of females. The PAS (Periodic acid-Schiff) and bright Coomassie blue positive reactions of the epithelium of the spermatic duct point to the secretion of polysaccharides and proteins that could promote the bundling of the spermatozoa. The internal epithelium of the ovarian wall secretes polysaccharides, protein, and lipid compounds throughout the storage and spawning period. The acid nature of the ovarian fluid during the storage period may maintain the bundling of spermatozoa when they enter the ovary and may also inhibit sperm motility until the moment of fertilization. The polysaccharide granules that come from the cryptal epithelium into the cavity where spermatozoa are maintained may supply them with nutrients for the storage period. The presence of glucosaminoglycans on the surface of the sperm is probably related to the inhibition of spermatic motility produced by the acidic environment. They are absent in the spermatozoa located in the testicular ducts, relatively scarce in those of the duct of the copulating organ, and abundant in those within the intraovarian cryptal structures.     

Seminal vesicle secretion of African catfish, its composition, its behaviour in water and saline solutions and its influence on gamete fertilizability

The seminal vesicle secretion (SVS) of the African catfish, Clarias gariepinus, was investigated by analytical and experimental methods. SVS consists mainly of proteins and glycoproteins which are responsible for its viscous and sticky nature. The secretion contains also high activities of acid phosphatase, alkaline phosphatase, and proteases. These catabolic enzymes do not have functions in autolysis or liquefaction of SVS but are considered to eliminate aging spermatozoa from the proximal portions of seminal vesicle and from the spermatic duct. SVS of the African catfish is unstable in the environment relevant for natural spawning. When SVS was mixed with water, seminal plasma or different types of saline solutions its protein coagulated forming fibrous or granular particles of variable size within a few seconds. Pure SVS completely inhibited the motility as the sticky secretion hindered spermatozoa in free swimming. SVS had also a negative effect on sperm fertility, egg fertility, and sperm egg contact, as the fertilization was drastically suppressed in the presence of SVS. Basing on our analytical and experimental results we exclude that SVS has functions in stabilizing the viability of spermatozoa stored in the spermatic ducts or is an energy resource of spermatozoa. It also does not improve or stabilize the fertilization process and has no functions in adhering the eggs to substrates or in covering the eggs for mechanical protection or antibacterial defense. A function of SVS in the male and female communication during the prenuptial spawning behaviour is discussed.     

Ultrastructure du canal des glandes agrégées et flagelliformes d'Araneus diadematus Clerck (Araneae,Araneidae)

Summary The excretory ducts of the silk glands which produce the viscid spiral of the webs ofAraneus diadematus show a complex structure. The duct of aggregate glands consists of three superposed types of cells. Several connective layers cover large and irregular nodule-forming cells which are rich in glycogen and mitochondria surrounded by invaginations of the plasma membranes. The internal cells, whose apical poles are lined by a cuticular intima, would be quite ordinary if not for the fact that they often carry large vacuoles which seem to empty themselves by exocytosis. Activity in the nodule cells is perceived from variations in the glycogen level and from the appearance of the mitochondria. Internal cells of the duct, when within the posterior spinneret, gradually acquire the characteristics of absorbing cells.The duct of flagelliform glands consists of two types of cells. The external cells, bounded by a simple basal lamina, are rich in mitochondria, glycogen, and invaginations of the plasma membranes; their activity is shown by variations in glycogen level and the extent of the extracellular spaces. The internal cells show numerous mitochondria either at the apical or basal poles, variable glycogen levels, long microvilli, and signs of apical absorption by pinocytosis; the sub-cuticular layer of the intima is particularly thick.We propose a functional interpretation of the aspects described above, and discuss it in terms of recent data on the chemical composition of silks. The excretory ducts are held to modify, by their activity, the secretory products of both types of glands. Solutes, especially phosphate ions, cross both cells and intima and would enter the glue of the aggregate glands which then undergoes partial dehydration in the posterior spinnerets. The product of the flagelliform glands seems to all appearance dehydrated during its passage in the duct and up to about the half-way through the posterior spinnerets. The liquid would flow through an extracellular path below the apical septate junctions of the internal cells. This study therefore favours attributing important role to the excretory ducts of silk glands in the final phase of the formation of silk fibres by spiders.     

Ultrastructure and histochemistry of the testicular efferent duct system and spermiogenesis in Opistognathus whitehurstii (Teleostei, Trachinoidei)

 Testis organization and spermatogenesis, with the emphasis on spermiogenesis, in Opistognathus whitehurstii are described by ultrastructural and histochemical methods. The germinal epithelium is extremely reduced and restricted to the periphery of the testis, while most of the organ is occupied by a highly developed system of testicular efferent ducts. A semicystic type of spermatogenesis is observed and in the germinal epithelium spermatogenesis occurs only until the spermatidal stage. Young spermatids are released into the lumen of the testicular lobules and mature to sperm within the efferent duct system. The epithelial cells of these ducts are involved in protein and glycogen secretion and in phagocytosis of degenerating germ cells and residual bodies cast off by developing spermatids. On the basis of these functions, the testicular efferent duct system cells are considered to be homologous to the Sertoli cells. A correlation between a highly developed testicular efferent duct system and semicystic spermatogenesis is examined and a possible functional meaning of this apparently unusual mode of sperm production is proposed. Accepted: 18 March 1997     

Sperm Storage Structures in the Ovary of Helicolenus dactylopterus dactylopterus (Teleostei: Scorpaenidae): an Ultrastructural Study

The ultrastructure of ovarian sperm storage of Helicolenus dactylopterus dactylopterus is described, before and after the spawning period. The spermatozoa remain inside cryptal structures that are situated in the interlamellar gaps and are connected to the ovarian lumen by a duct. This complex forms a highly specialised structure. During the long storage period, crypts are richly vascularised. Their surrounding simple epithelia have intercellular junctions that may serve to protect the spermatozoa from the female immune system. At the moment during which insemination of mature oocytes occurs, the sperm may be expelled from cryptal structures by means of a spasmodic contraction. During the post spawning period, residual spermatozoa that remain in the crypts are eliminated by cryptal phagocytes. At the end of the process the crypts contain only an amorphous material.     

Ultrastructure of the testicular gland of Blennius pavo (Pisces,Teleostei)

Summary The testicular gland of Blennius pavo Risso, 1810 is an accessory sex organ situated ventrally, adjacent to the testis. Sperm have to cross it to reach the vas deferens. Changes in the fine structure of the gland were monitored monthly over a period of 1 year. The annual cycle is closely correlated to the reproductive cycle. Maximum synthetic activity and cell size are reached during late prespawning and early spawning. At that time the number of lipid vacuoles increases markedly. Vesicles of smooth endoplasmic reticulum ER, numerous Golgi cisternae and tubulovesicular mitochondria which are characteristics of steroid producing cells can be observed. Secretory vesicles are synthesized in the apical region of the cells and then released into the ducts of the gland. The content of lipids is maintained until the resting period. The supranuclear region of the cells contains accumulations of lamellar bodies that probably derive from lysosomal vacuoles. Fine structural characteristics suggesting the function of the testicular gland are discussed.     

The reproduction ofBlennius pavo (Teleostei,Blenniidae). I. Ovarial cycle,environmental factors and feeding

Blennius (Salaria) pavo is a very common coastal fish in the Mediterranean Sea. The development of the ovary was studied over a period of one year; seven stages of oogenesis are described. Ripe eggs are carried during the spawning season from Juni to August. The ovary shows a resting period of more than half a year (September to March). Environmental factors of the study area such as surface water temperature and daylength are described as well as the condition factors of male and female fish. Observations on food and behaviour during the winter months are also included.     

The karyotype of Axarus festivus (Say) and a comparison with species of Lipiniella (Chironomidae: Diptera)

Larvae of Axarus andLipiniella share several apparantly apomorphic features of the mouth-part structure, however adults ofLipiniella show possible relationships toDemeijerea andChironomus. In order to assist in refining the relationships ofAxarus toLipiniella the karyotype ofAxarus festivus was determined and compared to the karyotypes ofLipiniella arenicola andL. moderata. Larvae ofA. festivus from a population in Kansas were monomorphous, with 2n=8, the Ist, IInd and IIIrd chromosomes metacentric, and IVth acrocentric.Axarus festivus therefore differs fromL. arenicola in chromosome number (2n=6), however homologous sections of all chromosomes were identified. Inversions were detected in the Ist, IInd and IIIrd chromosomes ofA. festivus relative toL. arenicola. It was determined that both species have high functional activity, as indicated by the presence of three Balbiani rings, and more than one nucleolus per genome. Differing degrees of polyteny, a feature previously described forL. arenicola, were observed in the salivary glands, with highest degrees of polyteny in cells near the salivary duct. These similarities of chromosome structure indicate close genetic relationships betweenA. festivus andL. arenicola. However, we did not find evidence for similarity ofA. festivus toL. moderata, which supports the previous conclusions byKiknadze et al. (1989) regardingL. arenicola andL. moderata.     

Histomorphological studies of the testis and male genital ducts of Supachai's caecilian,Ichthyophis supachaii Taylor, 1960 (Amphibia: Gymnophiona)

We investigated the structure of the male reproductive system in Ichthyophis supachaii. The testis comprises a series of mulberry‐like lobes, each of which contains testis lobules occupied by germ cysts. A single cyst consists of synchronously developing germ cells. Six spermatogenic cell types, viz. primary spermatogonia, secondary spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids and spermatozoa, have been identified and described. Notably, the testis of I. supachaii encompasses specific organization patterns of spermatids and spermatozoa during spermiogenesis. Spermiating cysts rupture and release spermatozoa to the collecting ducts, which are subsequently transported to the sperm duct, Wolffian duct and cloaca. We report for the first time ciliated cells in the epithelium of the caecilian Wolffian duct. The cloaca is divided into the urodeum and phallodeum. The urodeum has ciliated and glandular epithelia at its dorsolateral and ventral regions, respectively, as the lining of its internal surface. The muscular phallodeum is lined by ciliated epithelium. Paired Mullerian ducts lie parallel to the intestine and join the cloaca. The posterior portion of the duct is modified as the Mullerian gland. The most posterior region is non‐glandular and lined by ciliated epithelium. Our findings contribute further to information on the reproductive biology of caecilians in Thailand.     

Seasonal cell differentiation and olfactory potency of secretions by the anal glands of male peacock blenny Salaria Pavo

The current study shows cytological differences between the anal glands of pre‐spawning and spawning peacock blennies Salaria pavo. The cytological differences in the anal gland coincide with higher olfactory potency of their secretions in spawning males, suggesting that seasonal differentiation of cell clusters secreting neutral mucins is related to the production and release of the putative sex pheromone.     

Length–weight relationships for 22 fish species of the Mar Menor coastal lagoon (western Mediterranean Sea)

Length–weight relationships were estimated for 22 species of the Mar Menor coastal lagoon (south‐eastern Spain): Lipophrys dalmatinus, Salaria pavo, Callionymus pusillus, Aphanius iberus, Engraulis encrasicolus, Gobius cobitis, G. niger, G. paganellus, Pomatoschistus marmoratus, Symphodus cinereus, Chelon labrosus, Liza aurata, L. ramado, L. saliens, Mugil cephalus, Diplodus puntazzo, D. sargus sargus, Lithognathus mormyrus, Sarpa salpa, Sparus aurata, Hippocampus guttulatus and Syngnathus abaster. The captures were made in shallow areas of the lagoon during 2002–2004. Significant length–weight relationships were found for all species.     

Observations on the reproductive and larval biology ofBlennius pavo (Pisces: Teleostei)

Social behaviour and spawning of adultBlennius pavo kept in the laboratory are described. Eggs are deposited in batches on the walls of artificial spawning places (PVC pipes). One male guards and tends the eggs of different females in one spawning place. Larval hatching occurs in groups according to oviposition. Minimum incubation temperature is around 14–15°C. Larval survival in 1-1 rearing jars is not related to larval total length but to density of larval stock. An experimental population of laboratory reared juvenile and adolescentB. pavo displays a male to female ratio of 1:1.4. Factors possibly influencing the sex ratio of this littoral fish are discussed in view of the situation in its natural environment.     

Morphological and ecological investigations on sympatricLipophrys species (Blenniidae,Pisces)

The three syntopic blenniidsLipophrys canevai, L. adriaticus, andL. dalmatinus were investigated off Katarina Island near Rovinj, Yugoslavia, in regard to their habitats and food organisms. The shallow, sheltered rocky sea shore turned out to be the characteristic habitat forL. dalmatinus andL. adriaticus, whereasL. canevai also inhabits surf-exposed biotopes. The overlap of the microhabitats was below 50 %. Algae are the main food of all three species with respect to biomass, but harpacticoids dominated inL. adriaticus and especiallyL. dalmatinus when considering abundance of food organisms. Overlap of trophic niches was high (80 %) betweenL. canevai andL. adriaticus but lower (<45 %) betweenL. dalmatinus and the other two species. Morphological studies considered external morphology, pigmentation, and anatomy of eyes, including the closely related, cave-dwellingL. nigriceps. This species reveals special adaptations to life in dimly lit biotopes: reduction of the basic melanophore pigmentation, development of clinging organs, and enlargement of eyes and lenses combined with an effective ratio of retinal elements and modes of accommodation. It is concluded that the four species are members of one Lebensorttyp (Riedl, 1966) of whichL. nigriceps has superimposed the basic characteristics with specializations to a greater and the other three species to a lesser degree.     

Resorption of unemitted gametes in Lithognathus mormyrus (Sparidae,Teleostei): a possible synergic action of somatic and immune cells

The resorption of unemitted gametes during the post-spawning period of the male and female reproductive cycles in Lithognathus mormyrus was studied by histochemical, histological and cytological methods. The resorption of residual spermatozoa involved the phagocytotic activity of Sertoli cells bounding the seminiferous cysts of spermatozoa, and those associated with spermatogonia lining the lobular lumen. Spermatozoa remaining in the sperm duct were phagocytozed by the lining epithelial cells. Eosinophilic granulocytes and macrophages were identified in the vicinity of residual spermatozoa. The remnants of oocytes underwent an atretic phenomenon in which follicle cells were firstly involved, inducing a progressive fragmentation of the oocyte cytoplasm. Subsequently, eosinophilic granulocytes invaded oocyte degenerative areas and clung to the remaining vitelline inclusions ensuring their biotransformation into waste products (brown bodies). The analogy of the resorption processes of both male and female unemitted gametes during the post-spawning period of natural reproductive cycle, involving first the enveloping somatic cells and then immune cells, is emphasized.     

Sperm storage and degradation in the ovary of a marine copulating sculpin,Alcichthys alcicornis (Teleostei: Scorpaeniformes): Role of intercellular junctions between inner ovarian epithelial cells

The morphology of female sperm storage during the spawning period and the morphology of sperm degradation after the spawning period were investigated by electron microscopy in a copulating teleost, Alcichthys alcicornis. The spermatozoa were maintained in the ovarian cavity, floating in the ovarian fluid during the spawning period. The spermatozoa then degenerated and were phagocytized by macrophages invading the ovarian cavity after the spawning period. In the ovary during the spawning period, horseradish peroxidase used as a tracer revealed tight junctional complexes connecting adjacent cells of the inner ovarian epithelia (ovarian wall epithelium and ovigerous lamella epithelium). This indicates that a compartmentalization of the ovarian cavity occurs during the spawning period. The junctional complexes were breached after the spawning period, as shown by the fact that horseradish peroxidase penetrated the ovarian cavity via the intercellular space between the adjoining ovigerous lamella epithelia. These results suggest that the spermatozoa in the ovarian cavity are isolated from the maternal immune system by the tight junctional complexes between the adjoining inner ovarian epithelia during the spawning period, and then are eliminated by immune cells following the breakdown of the junctional complexes after the spawning period. J. Morphol. 233:153–163, 1997. © 1997 Wiley-Liss, Inc.