Phylogeny, expression and enzyme activity of zebrafish cyp19 (P450 aromatase) genes

The cyp19 encodes P450 aromatase, the enzyme catalyzing the conversion of estrogens from androgens. Estrogens affect the dimorphic, anatomical, functional and behavioral aspects of development of both males and females. In zebrafish, two cyp19 genes, cyp19a and cyp19b were found. They are expressed in ovary and brain, respectively. Expression of cyp19b can be detected by 11 days post-fertilization (dpf) by in situ hybridization in the olfactory bulbs, ventral telencephalic region and the hypothalamus of the brain in both male and female, where it is generally known to be affecting the reproductive function and sexual behavior. COS-1 clones permanently expressing the enzymes have been isolated. Both aromatase enzymes encoded by these two genes are functional in COS-1 cells and they can use androstenedione and testosterone equally efficiently. The presence of two functional cyp19 in zebrafish has its evolutionary and physiological importance.     

Sexually dimorphic and ontogenetic expression of dmrt1, cyp19a1a and cyp19a1b in Gobiocypris rarus

Fish have diverse sex determination and differentiation. DMRT1 and aromatase are conserved in the phyla and play pivotal roles in sex development. Gobiocypris rarus is a small fish used as a model in aquatic toxicology in China and has been used to study the effects of environmental endocrine disruptors on gene expression, but its sexual development remains elusive. Here, we report the full-length cDNA of G. rarus dmrt1 and its expression along with the expression of cyp19a1a and cyp19a1b, two genes encoding gonad and brain type aromatases, in adults and during ontogenesis. Both cyp19a1a and dmrt1 are expressed in the ovary and testis but show sexual dimorphism. Expression of cyp19a1a in the ovary is higher than in testes and dmrt1 follows the opposite pattern. Juvenile gonad histology changes at 15 days after hatching. The dimorphic expression of dmrt1 and cyp19a1a appears from 5 days after hatching, which is earlier than histological change. cyp19a1b is expressed coordinately with cyp19a1a until 15 days after hatching. These results show that dmrt1 and cyp19a1a play important roles in sex determination and sex differentiation in G. rarus.     

Isolation and characterization of 12 novel DNA microsatellites in the western rainbowfish, Melanotaenia australis

Microsatellites were isolated from the western rainbowfish, Melanotaenia australis, for use as molecular markers. Twelve polymorphic loci were found. When characterized using 32 individuals, these had between two and 24 alleles each, and observed heterozygosities ranged from 0.00 to 0.97. Ten loci were in Hardy-Weinberg equilibrium, and all of them showed independent inheritance. These loci will be useful for the study of molecular subdivision in this species.     

Regional differences in sexually dimorphic protein expression in the spontaneously hypertensive rat (SHR)

Hypertension is sexually dimorphic and modified by removal of endogenous sex steroids. This study tested the hypothesis that endogenous gonadal hormones exert differential effects on protein expression in the kidney and mesentery of SHR. At ~5 weeks of age male and female SHR underwent sham operation, orchidectomy, or ovariectomy (OVX). At 20-23 weeks of age, mean arterial pressure (MAP) was measured in conscious rats. The mesenteric arterial tree and kidneys were collected, processed for Western blots, and probed for Cu Zn superoxide dismutase (SOD1), soluble epoxide hydrolase (sEH), and Alpha 2A adrenergic receptor (A2AR) expression. MAP was unaffected by ovariectomy (Sham 164 ± 4: Ovariecttomy 159 ± 3 mm Hg). MAP was reduced by orchidectomy (Sham 189 ± 5:Orchidectomy 167 ± 2 mm Hg). In mesenteric artery, SOD1 expression was greater in male versus female SHR. Orchidectomy increased while ovariectomy decreased SOD1 expression. The kidney exhibited a different pattern of response. SOD1 expression was reduced in male compared to female SHR but gonadectomy had no effect. sEH expression was not significantly different among the groups in mesenteric artery. In kidney, sEH expression was greater in males compared to females. Ovariectomy but not orchidectomy increased sEH expression. A2AR expression was greater in female than male SHR in mesentery artery and kidney. Gonadectomy had no effect in either tissue. We conclude that sexually dimorphic hypertension is associated with regionally specific changes in expression of three key proteins involved in blood pressure control. These data suggest that broad spectrum inhibition or stimulation of these systems may not be the best approach for hypertension treatment. Instead regionally targeted manipulation of these systems should be investigated.     

Melanotaenia iris,a new freshwater rainbowfish (Melanotaeniidae) from Papua New Guinea with notes on the fish fauna in Head Waters

A new melanotaeniid rainbowfish,Melanotaenia iris, is described from mountain tributaries of the Strickland River in west-central Papua New Guinea. It is most closely related toM. goldiei of southern New Guinea, but differs in colour pattern, a distinctive high number of scales on the suborbital-preopercle bones, and a greater number of soft dorsal fin rays. Notes are also included on the poorly developed fish fauna of head waters in the Fly-Strickland and Sepik river systems. Most streams thus far investigated have less than 4 species. The plotosid catfishTandanus equinus appears to be the most successful colonizer of fast-flowing highland streams in the Fly-Strickland system and is sometimes the only species present.     

Sex- and tissue-specific expression of P450 aromatase (cyp19a1a) in the yellowtail clownfish,Amphiprion clarkii

To investigate the role of estrogen in the gonad of yellowtail clownfish Amphiprion clarkii, we isolated cDNA encoding cytochrome P450 aromatase (Cyp19a1a) from the adult ovary. The full-length cDNA of clownfish cyp19a1a is 1928-bp long and encodes 520 amino acids. Real-time quantitative RT-PCR analysis showed that cyp19a1a was expressed mainly in the ovary of female-phase fish. In situ hybridization and immunohistochemical observations showed that positive signals were restricted to the ovarian follicle of the female-phase fish. In contrast, Cyp19a1a signal was not detected in the ambisexual gonad of the male-phase fish. These findings suggest that Cyp19a1a is involved in oogenesis in the female-phase fish, but not in the ambisexual gonad of male-phase fish.     

Ultrastructure of the sexually dimorphic tarsal glands and tegumental glands in gonyleptoid harvestmen (Opiliones,Laniatores)

In at least four closely related families of the diverse harvestmen lineage Gonyleptoidea, males may possess sexually dimorphic tarsal glands in the swollen tarsomeres of the basitarsus and/or metatarsus of leg I. The first histological and ultrastructural examination of the sexually dimorphic tarsal glands in leg I focused only on Manaosbiidae. In this study, we examine the morphology and ultrastructure of the sexually dimorphic glands, and their associated glandular openings, found in the basitarsus and/or metatarsus of leg I of males representing Cosmetidae, Gonyleptidae, and Cranaidae (glandular openings only). In cosmetids and gonyleptids, the tarsal glands are made up of 20–60 glandular units that form distinct groups within the prolateral and retrolateral half of the tarsomere. Each glandular unit consists of a pair of terminal secretory cells, an intercalary cell wrapped around the receiving canal, and a canal cell tightly wrapped around the length of the conducting canal. Cosmetidae, Gonyleptidae, and Cranaidae exhibit remarkably similar tarsal glands and gland openings although the location of the glands in the leg differs slightly among them. Males of these three families exhibit markedly different glands and glandular openings compared to males of the family Manaosbiidae. The sexually dimorphic tarsal glands may provide an important morphological character for determining phylogenetic relationships among gonyleptoid families. Finally, we provide morphological and ultrastructural data for the common tegumental glands. These data indicate that the sexually dimorphic tarsal glands are strikingly similar to, and may possibly be derived from, the tegumental glands. J. Morphol. 274:1203–1215, 2013. © 2013 Wiley Periodicals, Inc.     

Anti-bacterial function in the sexually dimorphic pollinator rewards of Clusia grandiflora (Clusiaceae)

Many species of the dioecious, neo-tropical plant genus Clusia secrete a viscous, hydrophobic resin from glandular tissues in both male and female flowers. This substance is readily gathered by meliponine and euglossine bees for whom it most often serves as the sole pollinator reward. Bees use Clusia resin as a nest-building material. As such, resin clearly serves an indispensable mechanical function. However, resins with antimicrobial properties may also serve to reduce the risk of pathogenesis in the nest. If resin-gathering apids benefit from antimicrobial properties in nesting materials and are able to discern these characteristics in the forage they gather, one might predict that the resin reward presented in Clusia could have evolved under selection for both mechanical and antimicrobial properties. In dioecious species, where females and males each present a resin reward, selection regimes may differ between the sexes with the result that resin form and function diverge. We investigated both the form and function of the male and female pollinator reward resins of Clusia grandiflora. Using thin-layer chromatography (TLC), we compared the chemical compositions of floral resins from five widely separated populations of this species growing in southeastern Venezuela. We found that male and female resins exhibited a marked chemical dimorphism, with females having two major TLC-resolvable fractions and males having seven. This dimorphism was stable: there were no component differences between populations in either sex. Using a disk-diffusion technique, we surveyed the same resins for antimicrobial activity using assay microorganisms isolated from eusocial meliponine bees. Both male and female Clusia grandiflora resins had pronounced but relatively directed antimicrobial activity: both were toxic to 10 of 11 Gram-positive bacteria, 7 of 15 Gram-negative or variably-staining bacteria, 0 of 3 yeasts, and 0 of 3 filamentous fungi. Again with the disk-diffusion technique, we performed more detailed tests of resin bioactivity using two Gram-positive honeybee associates, Paenibacillus larvae and P. alvei, as model pathogens. Both male and female C. grandiflora resins were highly toxic to these honeybee pathogens. Female resin, however, produced zones of inhibition with more than twice the mean diameter of those produced by the male resin. This divergence in form and function of the C. grandiflora pollinator reward resins could be in response to different selective regimes as mediated by the pollinating insects. Received: 19 October 1998 / Accepted: 9 February 1999     

Evaluation of the toxicity of eggs,hatchlings and tadpoles of the introduced toad Bufo marinus (Anura: Bufonidae) to native Australian aquatic predators

Abstract The early life history stages of anurans in the Family Bufonidae often possess chemicals that are noxious or toxic to predators. Predators with no evolutionary history of exposure to bufomds may be particularly susceptible to these toxins. We conducted a series of laboratory experiments to investigate the toxic effects of eggs, hatchlings and tadpoles of the introduced toad, Bufo marinus (Linnaeus), on native Australian aquatic predators. There was considerable interspecific and intraspecific variation in these effects. Bufo marinus were highly toxic to some predator species, but were readily consumed by other species without apparent ill effect. Interspecific variation in toxic effects was not related to predator feeding mode or the number of B. marinus ingested by predators, and there was no clear pattern of distribution of vulnerability among species within higher taxa. Intraspecific variation in responses to toxins may result from individual variation in the resistance of predators to B. marinus toxins, or from individual variation in toxicity among B. marinus. Some native species adversely affected by B. marinus appeared unable to detect and avoid B. marinus toxins. This may result from a general inability to assess the toxicity of food items or from a lack of evolutionary exposure to B. marinus toxins.     

Molecular characterization and expression of cyp19a gene in Carassius auratus

A cyp19a gene that contains nine exons and eight introns was identified from Carassius auratus and was mainly expressed in the ovary. The cyp19a mRNA level after hatching was initially low, but began to increase from 25 days after hatching. A number of cis‐acting elements, such as the oestrogen receptor, steroidogenic factor 1 and SOX‐5 recognition sites, were found in the promoter of the cyp19 gene, which possesses a promoter function confirmed by a recombination green fluorescent protein checking system in vitro.     

Body size variation in the sexually dimorphic scaphopod Rhabdus rectius (Carpenter, 1864) (Dentaliida: Rhabdidae)

ABSTRACT

Male-biased sexual size dimorphism typically evolves via sexual selection for larger males that are favoured by choosy females or are more successful in mate competition with other males. Among marine invertebrates that broadcast their gametes into the ocean for fertilisation, this form of sexual size dimorphism is rare because such species lack direct interactions among males or between the sexes. However, the broadcast-spawning tusk shell Rhabdus rectius was recently reported to show strong male-biased sexual size dimorphism. That pattern might imply interesting and undiscovered sexual selection in this species. We found instead that the distribution of body size variation (weight, shell length) was similar between males and females of R. rectius, and mean sizes were not different between the sexes. However, we noted a male-biased sex ratio (～1:1.3) in our large sample of individuals. Many live scaphopods (and several dead shells) showed partial or complete boreholes drilled by predatory gastropods. Boreholes were observed on males and females in similar proportions. We collected scaphopods along with multiple individuals of one likely scaphopod predator, the small moon snail Euspira pallida, and in the lab we observed successful attacks by moon snails on tusk shells.