Differential regulation of msx genes in the development of the gonopodium,an intromittent organ,and of the "sword," a sexually selected trait of swordtail fishes (Xiphophorus)

The possession of a conspicuous extension of colored ventral rays of the caudal fin in male fish of swordtails (genus Xiphophorus) is a prominent example for a trait that evolved by sexual selection. To understand the evolutionary history of this so-called sword molecularly, it is of interest to unravel the developmental pathways responsible for extended growth of sword rays during development of swordtail males. We isolated two msx genes and showed that they are differentially regulated during sword outgrowth. During sword growth in juvenile males, as well as during testosterone-induced sword development and fin ray regeneration in the sword after amputation, expression of msxC is markedly up-regulated in the sword forming fin rays. In contrast, msxE/1 is not differentially expressed in ventral and dorsal male fin rays, suggesting a link between the development of male secondary sexual characters in fins and up-regulation of msxC expression. In addition, we showed that msx gene expression patterns differ significantly between Xiphophorus and zebrafish. We also included in our study the gonopodium, a testosterone-dependent anal fin modification that serves as a fertilization organ in males of live-bearing fishes. Our finding that increased levels of msxC expression are associated with the testosterone-induced outgrowth of the gonopodium might suggest either that at least parts of the signaling pathways that pattern the evolutionary older gonopodium have been coopted to evolve a sexually selected innovation such as the sword or that increased msxC expression may be inherent to the growth process of long fin rays in general.     

Testis and gonopodium development in Anableps Dowi (Pisces: Anablepidae) correlated with pituitary gonadotropic zone area

Testicular development in the viviparous fish, Anableps dowi, is described from embryonic stages through sexual maturation and is correlated with development of the gonadotropic zone of the pituitary. Only isolated spermatogonia or cysts of early spermatogonia are found in embryos 1.8 to 4.4 cm in length and postnatal specimens 5.4 to 8.8 cm in length. Cysts with more advanced spermatogonia are seen in specimens from 7.9 to 10.8 cm in length. Spermatogenetic meioses are first observed in a 10.2 cm male. Specimens greater than 12.0 cm have mature spermatozoa (partial spermatozeugmata) within the main testicular ducts, indicating gonadal maturation. The morphological transformation of the anal fin into the intromittent gonopodium is also described, using both whole and cleared specimens. Even in the smallest postnatal male available (5.4 cm), the anal fin can be distinguished from that of an equivalent sized female. Anal fin changes proceed until the mature gonopodium is developed, when body lengths greater than 16 cm are reached. Testicular maturation, defined as the presence of partial spermatozeugmata in the main testicular ducts, precedes the completion of gonopodial development. Measurements of ventral gonadotropic zone area in pituitary mid-sagittal sections show a continuous increase from neonatal through sexually mature specimens. The results point to a role for testicular androgens in early testis development.     

Gonopodial system review and a new fish record of Poeciliopsis infans (Cyprinodontiformes: Poeciliidae) for Lake Patzcuaro,Michoacan, central Mexico

Since 1997, Poeciliopsis infans Woolman 1894 has been recognized as a new inhabitant of Lake Patzcuaro, Michoacan in Central Mexico. Between February 1997 and October 1998, nine fish samples were collected at Lake Patzcuaro. Morphometric and meristic counts were conducted on a random selection of 40 organisms of both sexes of Poeciliopsis infans. Males of these viviparous fish posses a modified anal fin called gonopodium. The characteristic hemal spine on the 18th caudal vertebra for this species is described herein and the bony components of the gonopodial structure and suspensoria that together comprise the gonopodial system, which is important for taxonomic studies at various levels of classification were reviewed. Poeciliopsis infans displays a high degree of sexual dimorphism in body shape and anal fin anatomy with the most conspicuous difference observed in anal fin height, which averages 40% of SI in males and 17% in females. Comparisons between male and female anal fins are described herein as well as the possible impacts of this species on Lake Patzcuaro fish fauna.     

Androgen dependent development of a modified anal fin, gonopodium, as a model to understand the mechanism of secondary sexual character expression in vertebrates

Male external genitalia show structural variations among species. Androgenic hormones are essential for the morphological specification of male type copulatory organs, while little is known about the developmental mechanisms of such secondary sexual characters. Western mosquitofish Gambusia affinis may offer a clue to the sexual differentiation researches, because they show a prominent masculine sexual character for appendage development, anal fin to gonopodium (GP) transition, and its formation could be induced in early juvenile fry by exogenously supplied androgens. We show that GP development is promoted by androgen dependent augmentation of sonic hedgehog (Shh) expression. Two AR cDNAs were cloned and identified as ARalpha and ARbeta from western mosquitofish. Both ARs were predominantly expressed in the distal region of outgrowing anal fin rays. Exposure of fry to androgen caused anal fin outgrowth concomitant with the Shh induction in the distal anal fin ray epithelium. When AR signaling was inhibited by its antagonist flutamide in fry, the initial induction of the Shh was suppressed accompanying retarded anal fin outgrowth. Similar suppression of anal fin outgrowth was induced by treatment with cyclopamine, an inhibitor of Shh signaling. These observations indicate that androgen dependent Shh expression is required for anal fin outgrowth leading to the formation of a genital appendage, the GP in teleost fishes. Androgen-induced GP formation may provide insights into the expression mechanism regulating the specification of sexual features in vertebrates.     

Retinoic acid regulation by CYP26 in vertebrate lens regeneration

Xenopus laevis is among the few species that are capable of fully regenerating a lost lens de novo. This occurs upon removal of the lens, when secreted factors from the retina are permitted to reach the cornea epithelium and trigger it to form a new lens. Although many studies have investigated the retinal factors that initiate lens regeneration, relatively little is known about what factors support this process and make the cornea competent to form a lens. We presently investigate the role of Retinoic acid (RA) signaling in lens regeneration in Xenopus. RA is a highly important morphogen during vertebrate development, including the development of various eye tissues, and has been previously implicated in several regenerative processes as well. For instance, Wolffian lens regeneration in the newt requires active RA signaling. In contrast, we provide evidence here that lens regeneration in Xenopus actually depends on the attenuation of RA signaling, which is regulated by the RA-degrading enzyme CYP26. Using RT-PCR we examined the expression of RA synthesis and metabolism related genes within ocular tissues. We found expression of aldh1a1, aldh1a2, and aldh1a3, as well as cyp26a1 and cyp26b1 in both normal and regenerating corneal tissue. On the other hand, cyp26c1 does not appear to be expressed in either control or regenerating corneas, but it is expressed in the lens. Additionally in the lens, we found expression of aldh1a1 and aldh1a2, but not aldh1a3. Using an inhibitor of CYP26, and separately using exogenous retinoids, as well as RA signaling inhibitors, we demonstrate that CYP26 activity is necessary for lens regeneration to occur. We also find using phosphorylated Histone H3 labeling that CYP26 antagonism reduces cell proliferation in the cornea, and using qPCR we find that exogenous retinoids alter the expression of putative corneal stem cell markers. Furthermore, the Xenopus cornea is composed of an outer layer and inner basal epithelium, as well as a deeper fibrillar layer sparsely populated with cells. We employed antibody staining to visualize the localization of CYP26A, CYP26B, and RALDH1 within these corneal layers. Immunohistochemical staining of these enzymes revealed that all 3 proteins are expressed in both the outer and basal layers. CYP26A appears to be unique in also being present in the deeper fibrillar layer, which may contain cornea stem cells. This study reveals a clear molecular difference between newt and Xenopus lens regeneration, and it implicates CYP26 in the latter regenerative process.     

Gonopodial morphogenesis in female mosquitofish,Gambusia affinis affinis,masculinized by exposure to degradation products from plant sterols

Synopsis Female mosquitofish,Gambusia affinis affinis, were masculinized by exposure to degradation products (presumably steroids) of the plant sterol, stigmastanol. Masculinization was indicated by the induction of a male-like gonopodium in each specimen. The morphogenetic stages of gonopodial development are discussed as are the anatomical specializations produced in the mature gonopodial tip.     

Fibroblast growth factor pathway component expression in the regenerating zebrafish fin

Adult zebrafish regenerate their appendages (fins) after amputation including the regeneration of bone structures (fin rays). Fibroblast growth factor (Fgf) signaling, which is involved in morphogenetic processes during development, has been shown to be essential for the process of fin regeneration. Moreover, mutations in Fgf pathway component genes lead to abnormal skeletal growth in teleosts and mammals, including humans, illustrating the importance of Fgf signaling in the growth control of tissues. Here, we revisited Fgf signaling pathway component expression by RNA in situ hybridization to test for the expression of about half of the ligands and all receptors of the pathway in the regenerating zebrafish fin. Expression patterns of fgf7, fgf10b, fgf12b, fgf17b and fgfr1b have not been reported in the literature before. We summarize and discuss known and novel localization of expression and find that all five Fgf receptors (fgfr1a, fgfr1b, fgfr2, fgfr3 and fgfr4) and most of the tested ligands are expressed in specific regions of the regenerate. Our work provides a basis to study domain specific functions of Fgf signaling in the regenerating teleost appendage.     

Two new genera and species of the subfamily Brosmophycinae (Bythitidae,Ophidiiformes) from Northern Australia

Two new bythitid genera and species of the subfamily Brosmophycinae are described from Northern Territory, Australia.Brosmolus longicaudus, described from a single male specimen, is unique in the tribe Brosmophycini in having the anal fin origin well anterior to the midpoint of the body and thin, transparent skin on the head and body.Beaglichthys macrophthalmus, described from a single female specimen, differs from all other genera in the subfamily by the following combination of characters: eight branchiostegal rays, eye diameter longer than snout length, cheek scaly, anal fin origin at midpoint of body, three developed rakers on the first gill arch, 12 caudal fin rays, and 14 precaudal vertebrae.     

Combinatorial roles for zebrafish retinoic acid receptors in the hindbrain, limbs and pharyngeal arches

Retinoic acid (RA) signaling regulates multiple aspects of vertebrate embryonic development and tissue patterning, in part through the local availability of nuclear hormone receptors called retinoic acid receptors (RARs) and retinoid receptors (RXRs). RAR/RXR heterodimers transduce the RA signal, and loss-of-function studies in mice have demonstrated requirements for distinct receptor combinations at different stages of embryogenesis. However, the tissue-specific functions of each receptor and their individual contributions to RA signaling in vivo are only partially understood. Here we use morpholino oligonucleotides to deplete the four known zebrafish RARs (raraa, rarab, rarga, and rargb). We show that while all four are required for anterior-posterior patterning of rhombomeres in the hindbrain, there are unique requirements for rarga in the cranial mesoderm for hindbrain patterning, and rarab in lateral plate mesoderm for specification of the pectoral fins. In addition, the alpha subclass (raraa, rarab) is RA inducible, and of these only raraa expression is RA-dependent, suggesting that these receptors establish a region of particularly high RA signaling through positive-feedback. These studies reveal novel tissue-specific roles for RARs in controlling the competence and sensitivity of cells to respond to RA.     

A new horned sculpin (Pisces: Cottidae) from the Miocene of Sakhalin Island,Russia

A new species of the Recent genus Enophrys, E. hoplites sp. nov., is described from the Miocene (Serravalian?Tortonian) Agnevo Formation of Sakhalin Island, Russia. This Miocene species differs from two Recent congeners in the greater number of anal fin rays (14 versus 9?13), the smaller head, less raised margins of the orbit, flatter upper contour of the head, longer anal fin base, short predorsal distance, higher first dorsal fin, and in the right angle (versus acute angle) between two upper preopercular spines. An archaic feature of the new species is the presence of two rays on the posterior pterygiophore of the anal fin, a character recorded only in the most primitive Recent members of the family. The occurrence of this new fossil species shows that extant subfamilies, tribes, and genera existed as a part of the family Cottidae as early as the Miocene time.     

A new creediid fishCreedia bilineatus from the Yaeyama Islands,Japan

A new creediid fish,Creedia bilineatus, is described on the basis of 3 specimens from the Yaeyama Islands, Okinawa Pref., Japan. This new species is distinguished from all other species of the genusCreedia by having fewer anal fin rays (17–18 vs. 24–28).     

Systematic observations on tropical Asian medakas or ricefishes of the genusOryzias, with descriptions of four new species

The so-calledOryzias melastigma (McClelland, 1839), reported from India, Bangladesh, Myanmar, and Malaysia by numerous authors beginning with Day (1877), is based mainly or entirely onAplocheilus panchax (Hamilton, 1822). India and Bangladesh have two species ofOryzias, both large. The deeper-bodied of these two species is reported for the first time asO. dancena (Hamilton, 1822). The other is identified asO. carnaticus (Jerdon, 1849). Myanmar has one large species,O. dancena, and one tiny species,O. uwai new species.Oryzias minutillus Smith, 1945 andO. uwai differ from all otherOryzias in having 4/5 instead of 5/6 principal caudal fin rays.Oryzias uwai differs fromO. minutillus in being more conspicuously pigmented and having large, 6-rayed pelvic fins often extending to anal fin origin instead of much smaller and shorter 5-rayed pelvic fins. In Thailand (including its part of the Mekong basin) three species are known: a large estuarine species tentatively identified asO. javanicus (Bleeker, 1854) and two tiny inland species,O. mekongensis Magtoon & Uwa, 1986, andO. minutillus. Oryzias minutillus from many localities are more or less melanoproctic, i.e. have a darkly pigmented genital or vent area not seen in other species. The Mekong basin in Laos has two large species,O. latipes (Temminck & Schlegel, 1846) andO. pectoralis new species, distinguished by a prominent black blotch on the pectoral fin base, both recently collected in the Nam Theun watershed in central Laos; and two tiny species,O. mekongensis andO. minutillus. Only one species ofOryzias is known from the Mekong delta in Vietnam, the small moderately deep-bodiedO. haugiangensis new species, with 19–22 anal and 9–10 pectoral fin rays. The Indonesian island of Java has one large species,O. javanicus (Bleeker, 1852) with 21–25 anal and 11 pectoral fin rays, and one small species,O. hubbsi new species, with only 17–21 anal and 9 pectoral fin rays.     

New genus and species of the family stichaeidae from Hokkaido,Japan

Specimens of a new genus and species of the stichaeid fish,Leptostichaeus pumilus, were collected from the Okhotsk Sea off Hokkaido in Japan. The present new genus and species clearly differs from all the other genera and species of the stichaeid fishes in the following characters: 3 or 4 pectoral fin rays; 10 or fewer caudal principal rays; 79–82 dorsal spines; no pelvic fin; last interneural spine supporting a single dorsal spine; infraorbital, occipital and lateral line canals absent; moderate size of dorsal spine shorter than eye diameter; membranes of dorsal and anal fins widely connected with caudal fin; a large black spot divided by a yellow band present just above gill cover.     

Schindleria elongata,a new species of paedomorphic gobioid from the Red Sea (Teleostei: Schindleriidae)

A new species of paedomorphic gobioid, Schindleria elongata, from the Red Sea, is described on the basis of five specimens. The new species is characterized by its lack of body pigmentation; the body depth at pectoral‐fin origin 4–5% of standard length (L_S) and at anal‐fin origin 5–7% L_S; the predorsal length 66–70% L_S; pre‐anal length 66–71% L_S; dorsal‐fin rays 13 or 14; anal‐fin rays 10 or 11; first dorsal‐fin ray at myomere 20 or 21; first anal‐fin ray below second to fourth dorsal‐fin rays; myomeres 19 or 20 + 13 or 14 = 33 or 34 total; premaxillae and dentaries with small teeth; gas bladder located posteriorly at 56–60% L_S; males with a rod‐like, flexible urogenital papilla lacking lobes, projections or accessory papillae, with distal half tapering to a broad, angular point and usually posteriorly directed. A key to the species of Schindleriidae is presented.     

The median‐fin skeleton of the Eastern Atlantic and Mediterranean clingfishes Lepadogaster lepadogaster (Bonnaterre) and Gouania wildenowi (Risso) (Teleostei: Gobiesocidae)

Previous research on the osteology of the Gobiesocidae focused mostly on the neurocranium and the thoracic sucking disc (formed by the paired‐fin girdles). Little attention has been paid to the skeleton of the median fins. The dorsal‐ and anal‐fin skeleton of Lepadogaster lepadogaster and other gobiesocids (excluding Alabes, which lacks these fins) are characterized by the absence of spines, branched fin‐rays, and middle radials. In gobiesocids, the distal radials never ossify and consist of elastic hyaline‐cell cartilage. Gouania wildenowi is unique among gobiesocids in having further reductions of the dorsal‐ and anal‐fin skeleton, including a notable decrease in the size of the proximal‐middle radials in an anterior–posterior direction. Unlike L. lepadogaster, which exhibits a one‐to‐one relationship between the dorsal‐ and anal‐fin rays and proximal‐middle radials, G. wildenowi has a higher number of proximal‐middle radials than distal radial cartilages and fin rays in the dorsal and anal fins. In G. wildenowi, the dorsal‐ and anal‐fin rays do not articulate with the distal tip of the proximal‐middle radials but are instead positioned between proximal‐middle radials, which is unusual for teleosts. Previously unrecognized dorsal and ventral pads of elastic hyaline‐cell cartilage are also present in the caudal skeleton of L. lepadogaster, G. wildenowi, and all other gobiesocids examined. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

A new loach, Cobitis shikokuensis (Teleostei: Cobitidae), from Shikoku Island, Japan

A new species of spinous loach, Cobitis shikokuensis, is described based on 297 specimens from Shikoku Island, Japan. The new species was formerly known as the Shikoku group of Cobitis takatsuensis. It can be distinguished from other species of Cobitis and closely related genera by a combination of the following characters: dorsal fin with 6 branched soft rays; anal fin with 5 branched soft rays; one brownish streak across eye from the tip of nose, no streak on cheek; a black spot smaller than eye diameter near the dorsal corner of the caudal fin base; 3–5 small brownish speckles on ventral side of caudal peduncle; high caudal peduncle with well-developed fleshy keels on dorsal and ventral side; a lamina circularis at base of dorsal part of pectoral fin absent; first branched soft ray of pectoral fin broad in males; pectoral soft rays widely branched from the approximate midpoint; last anal fin ray with 2 elements; interorbital width 11.2–17.1% of head length.     

Neolumpenus unocellatus,a new genus and species of stichaeid fish from Japan

Neolumpenus unocellatus gen. et sp. nov., a stichaeid fish (subfamily Lumpeninae,sensu Makushok, 1958) is described on the basis of a single specimen found in the stomach of the Pacific cod,Gadus macrocephalus Tilesius, caught off Akkeshi, Hokkaido, Japan. The new genus and species is distinguished from all other lumpenines in having the following combination of characters: 1) 51 dorsal spines, 33 anal fin rays, 57 total vertebrae; 2) broad pelvic fin with deeply-branched soft rays; 3) lower rays of pectoral fin branched and not prolonged backward; 4) prevomerine and palatine teeth present; 5) pungent spines present in pelvic and anal fins; 6) upper lip fused to snout anteriorly; 7) gill openings not extending forward beyond a vertical through posterior margin of eye; 8) minimal (fifth) hypural present; 9) first interneural spine inserted between first and second neural spines; 10) extremely large cephalic sensory pores present; 11) high, steep snout; 12) ocellus on dorsal base of caudal fin.