Identification of a novel C1q family member in color crucian carp (Carassius auratus) ovary

Potential roles of Clq/tumor necrosis factor (TNF) superfamily proteins have been observed in vertebrate oogenesis and oocyte maturation, but no ovary-specific member has been identified so far. In this study, we have cloned and identified a novel member of Clq family with a Clq domain in the C-terminal from fully grown oocyte cDNA library of color crucian carp and demonstrated that the gene might be specifically expressed in ovary and therefore designated as Carassius auratus ovary-specific Clq-like factor, CaOClq-like factor. It encodes a 213 amino acid protein with a 17 amino acid signal peptide. There is only one protein band of about 24.5 kDa in the extracts from phase I to phase IV oocytes, but two positive protein bands are detected in the extracts of mature eggs and fertilized eggs. Furthermore, the mobility shift of the smaller target protein band cannot be eliminated by phosphatase treatment, but the larger protein band increases its mobility on the gel after phosphatase treatment, suggesting that the larger protein might be a phosphorylated form. Immunofluorescence localization indicates that the CaOClq-like proteins localize in cytoplasm, cytoplasm membrane and egg envelope of the oocytes at cortical granule stage and vitellogenesis stage, whereas they were compressed to cytoplasm margin in ovulated mature eggs and discharged into perivitelline space between cytoplasm membrane and egg envelope after egg fertilization. Further studies on distribution and translocation mechanism of the CaOClq-like factor will be benefit to elucidate the unique function in oogenesis, oocyte maturation and egg fertilization. (C) 2004 Elsevier Inc. All rights reserved.

cDNA cloning and characterization of a novel SNX gene differentially expressed in previtellogenic oocytes of gibel carp

To understand the molecular events governing fish oogenesis, a multiple technique was used to identify the genes differentially expressed at different phases during fish oogenesis. This technique is a combination of suppression subtractive hybridization, SMART cDNA synthesis and RACE-PCR. Here we report the cDNA cloning and expression characterization of a novel SNX gene based on its differential transcription between previtellogenic and fully mature oocytes in naturally gynogenetic gibel carp. First, a cDNA fragment selectively expressed in previtellogenic oocytes was identified and used to screen a SMART cDNA library prepared from the same mRNA sample by RACE-PCR for cloning fully length cDNA. The full length cDNA was 1392-bp long and coded for a novel SNX protein with 225 amino acids. The 5' UTR had 72 bp and 3' UTR had 642 bp. Unlike most of maternal genes that are transcribed after vitellogenesis and stored in oocytes, this gene is expressed at a higher level in the previtellogenic oocytes and at a much lower level in fully matured oocytes. However, RT-PCR analysis of tissues showed it was ubiquitous transcription. The novel gene is named fish sorting nexin (fSNX), because it contains a conserved PX domain. The fact which major expression of the gene occurs in the previtellogenic oocytes suggests that it might have an important function in the oogenesis. (C) 2003 Elsevier Inc. All rights reserved.

Differential expression and characterization analysis of a new gene with WD domains in fish oogenesis

A new gene with WD domains is cloned and characterized according to its differential transcription and expression between previtellogenic oocytes (phase I oocytes) and fully-grown oocytes (phase V oocytes) from natural gynogenetic silver crucian carp (Carassius auratus gibelio) by using the combinative methods of suppressive subtraction hybridization, SMART cDNA synthesis and RACE-PCR. The full-length cDNA is 1870 bp. Its 5 ' untranslated region is 210 bp, followed by an open reading frame of 990 bp, which has the typical vertebrate initiator codon of ANNATG. The open reading frame encodes a protein with 329 amino acids. It has 670 bp of 3 ' untranslated region and an AATAAA polyadenylation signal. Because it has 92% homology to STRAP (serine-threonine kinase receptor-associated protein), a recently reported gene, we named it FSTRAP (fish STRAP). Virtual Northern blotting indicated that the FSTRAP was transcribed in fully-grown oocytes (phase V oocytes), but not in previtellogenic oocytes (phase I oocytes). RT-PCR analysis showed that FSTRAP was transcribed in brain, heart, kidney, muscle, ovary, spleen and testis, but not in liver. And its mRNA could be detected in the oocytes from phase II to phase V. Western blotting also showed that FSTRAP protein could be detected in brain, heart, kidney, muscle, ovary, spleen and testis except liver. Results of Western blotting on various oocytes were also similar to the RT-PCR data. FSTRAP protein was not expressed in the previtellogenic oocytes. Its expression initiated from phase II oocytes after vitellogenesis, and was consistent with the mRNA transcription.

Concentrations of alkyphenol polyethoxylates entering UK estuaries

Concentrations of the weakly oestrogenic degradation products of alkylphenol polyethoxylate (APE) surfactants (nonylphenol, octylphenol, nonylphenol monoethoxylate and nonylphenol diethoxylate) were measured in water and sediments from British rivers and estuaries collected during 1994 and 1995. In addition, a series of samples of tissues of wild fish from the River Aire, and from a laboratory dosing experiment were analysed for alkylphenols, to assess the degree of bioaccumulation of these compounds. Measurable concentrations of APE residues were recorded in the River Aire (15–76 μg/l total extractable alkylphenols), the River Mersey (6–11 μg/l) and the Tees estuary (up to 76 μg/l). These levels exceed, or are close to, the no observed effect concentration for the induction of vitellogenesis in caged trout (5–20 μg/l total extractable alkylphenols), and may be sufficient to exert an oestrogenic effect on fish populations in these areas. A sediment sample from Bingley on the River Aire contained 15 μg/g (dry weight) nonylphenol, and concentrations in sediments from the Tees and Mersey estuaries exceeded 1 μg/g. These rivers receive a variety of trade waters via sewage treatment works (STW) effluents containing significant concentrations of APE. Elsewhere, concentrations in water and sediments were near or below limits of detection and biological effects are unlikely, suggesting that any oestrogenic effects observed in sewage outfalls and rivers not directly impacted by APE-containing trade-waters may be caused by other chemicals. Analysis of samples of trout muscle taken from a tank dosed at 65 μg/l nonylphenol indicated a bioaccumulation factor of between 90 and 125 after 3 weeks exposure. Samples of wild fish from the River Aire contained up to 0.8 μg/g nonylphenol in the muscle, a tissue bioaccumulation factor of approximately 50 relative to measured concentrations in water samples. A series of fish samples taken from offshore for food quality assurance purposes contained no detectable levels of APE residues (0.05–0.1 μg/g nonylphenol).

Inhibitory effects of the androgenic gland on ovarian development in the mud crab Scylla paramamosain

Isolation and characterization of androgenic hormone in decapod crustaceans depend on an effective bioassay of its action. In the present study, the effect of androgenic gland on ovarian development in the mud crab Scylla paramamosain was investigated with a view to develop a bioassay for androgenic hormone. Ovarian regression with degeneration of oocytes occurred in some female crabs implanted with androgenic gland in vivo. In vitro incubation of ovarian tissues at secondary vitellogenesis in extract of androgenic gland resulted in a significant decrease in amino acid uptake by the tissues. We propose that this inhibitory effect could be established as an effective bioassay for the isolation of androgenic hormone in the mud crab. (c) 2005 Published by Elsevier Inc.

The Influence of Male Diet on Life History Traits of Female Mosquitoes

In the field, mosquitoes characteristically feed on sugars soon after emergence and intermittently during their adult lives. Sugar meals are commonly derived from plant nectar and homopteran honeydew, and without them, adults can only survive for a few days on larval reserves. In addition to sugar, females of most species rely on blood for the initiation and maintenance of egg development; thus their reproductive success depends to some extent on the availability of blood hosts. Males, on the other hand, feed exclusively on sugars. Consequently, their sexual maturation and reproductive success is largely dependent upon access to sugar sources. Plant nectar and homopteran honeydew are the two main sugar sources utilized by mosquitoes in the wild. Previous laboratory studies had shown that differences between nectar sources can affect the survivorship and biting frequency of disease vectoring mosquitoes. However, little is known on how sugar composition influence the reproductive processes in male mosquitoes. Male mosquitoes transfer accessory gland proteins and other hormones to their mates along with sperm during mating. In the female, these seminal fluid constituents exert their influence on reproductive genes that control ovulation and vitellogenesis. The present study tests the hypothesis that the mates of males consuming different sugar meals will exhibit varying levels of induction of vitellogenin (a gene which regulates the expression of egg yolk precursor proteins). Real-time quantitative RT-PCR was used to investigate how each sugar meal indirectly influences vitellogenin mRNA abundance in female Anopheles stephensi following mating. Results indicate that mates of nectar-fed males exhibit 2-fold greater change in vitellogenin expression than the mates of honeydew-fed males. However, this response did not occur in non-blood fed controls. These findings suggest that the stimulatory effect of mating on vitellogenesis in blood meal-reliant (i.e. anautogenous) mosquitoes may only be synergistic in nature. The present study also sought to compare the potential fitness costs of mating incurred by females that do not necessarily require a blood meal to initiate a reproductive cycle (i.e., exhibit autogeny). Females of the facultatively autogenous mosquito, Culex molestus were allowed to mate with males sustained on either nectar or honedyew. Mean lifetime fecundity and survivorship of females under the two different mating regimes were then recorded. Additionally, one-dimensional gel electrophoresis was used to verify the transfer of male accessory gland proteins to the sperm storage organs of females during mating.While there was no significant difference in survival between the test treatments, the mates of nectar-fed males produced 11% more eggs on average than mates of honeydew-fed males. However, additional data are needed to justify the extrapolation of these findings to natural settings. These findings prompt further investigation as the differences caused by diet variation in males may be reflected across other life history traits such as mating frequency and insemination capacity.

Studies On The Reproduction Of Indian Whiting Sill/160 5'/Ham/1 (Forskal) (Percoidei, Sillaginidae)

From the point of view of rational exploitation and proper management of the fishery resources as well as for the development of intensive aquaculture of fishes through selective breeding, brood stock development, domestication and genetic improvement, a sound knowledge of reproductive biology and physiology of the candidate species is of great importance. In recent times, a wealth of information on maturity, spawning habits, spawning periodicity, spawning season, size at maturity and fecundity of commercially important fishes has been generated. Gametogenesis involves the transformation of Primordial germ cells in the gonads into specialised cells or gametes, namely ova in the female and sperms in male, through a series of complex morphological and cytological events. The formation of male gamete is known as spermatogenesis. In the female, the primary growth phase involving the formation of primary oocyte from oogonia is known as oogenesis, which would be followed by the secondary growth phase, in which considerable increase in the size of the oocyte occurs, due mainly to accumulation of yolk. This process is known as vitellogenesis, which would be followed by final maturation and ovulation of the ova. In the present work, basic aspects of maturation and spawning, salient features of gametogenesis and associated biochemical changes occurring during these processes in an important cultivable fish, Sillago sihama belonging to the family Sillaginidae have been investigated.

Reproductive ecology of dipsadine snakes, with emphasis on South American species

A relatively large amount of variation occurs in the reproductive ecology of tropical snakes, and this variation is generally regarded as being a consequence of seasonality in climate and prey availability. In some groups, even closely related species may differ in their reproductive ecology; however, in others it seems to be very conservative. Here we explore whether characters related to reproduction are phylogenetically constrained in a monophyletic group of snakes, the subfamily Dipsadinae, which ranges from Mexico to southern South America. We provide original data on reproduction for Leptodeira annulata, Imantodes cenchoa, and three species of Sibynomorphus from southern, southeastern and central Brazil, and data from literature for other species and populations of dipsadines. Follicular cycles were seasonal in Atractus reticulatus, Dipsa, albifrons, Hypsiglena torquata, Leptodeira maculata, L. punctata, Sibynomorphus spp. and Sibon sanniola from areas where climate is seasonal. In contrast, extended or continuous follicular cycles were recorded in Dipsas catesbyi, D. neivai, Imantodes cenchoa, Leptodeira annulata, and Ninia maculata from areas with seasonal and aseasonal climates. Testicular cycles also varied from seasonal (in H. torquiata) to continuous (in Dipsa,5 spp., Leptodeira annulata, L. maculata, N. maculata, and Sibynomorphus spp.). Most dipsadines are small (less than 500 rum SVL), and females attain sexual maturity with similar relative body size than males. Sexual dimorphism occurred in terms of SVL and tail length in most species, and clutch size tended to be small (less than five eggs). Combat behavior occurs in Imantodes cenchoa, which did not show sexual size dimorphism. Reproductive timing, for both females and males, varied among species but in general there were no differences between the tribes of Dipsadinae in most of the reproductive characteristics, such as mean body size, relative size at sexual maturity, sexual size and tail dimorphism, duration of vitellogenesis or egg-carrying in oviducts.

Fatty acids as trophic biomarkers in vitellogenic females in an impounded tropical river

Fatty acids have been used in marine biogeochemistry as food chain biomarkers, but in freshwater these studies are rare. In order to evaluate the fatty acid potential as biomarkers in freshwater, their profile was analyzed during vitellogenesis in two fish species, in both waterfall and reservoir environments of the Paraiba do Sul River Basin. Detrivorous Hypostomus affinis and omnivorous Geophagus brasiliensis seem to elongate and desaturate polyunsaturated fatty acids (PUFA) and transfer them to the ovaries` phospholipids. Waterfall Geophagus brasiliensis have more highly unsaturated fatty acids in the liver, but in the reservoir, accumulation mainly occurs in muscle and ovary triglycerides, suggesting trophic opportunism and a plasticity during vitellogenesis. In Hypostomus affinis, PUFA alteration occurs only in the reservoir, suggesting a high phytoplankton occurrence. Eutrophication and water speed is reflected in Hypostomus affinis ovaries by higher PUFAn3 and bacterial fatty acids. As in marine environments, analysis of mono- and polyunsaturated fatty acids during vitellogenesis can be used as a tool in food chain studies in freshwater.

Effects of Plasmodium gallinaceum on hemolymph physiology of Aedes aegypti during parasite development

Insect disease vectors show diminished fecundity when infected with Plasmodium. This phenomenon has already been demonstrated in laboratory models such as Aedes aegypti, Anopheles gambiae and Anopheles stephensi. This study demonstrates several changes in physiological processes of A. aegypti occurring upon infection with Plasmodium gallinaceum, such as reduced ecdysteroid levels in hemolymph as well as altered expression patterns for genes involved in vitellogenesis, lipid transport and immune response. Furthermore, we could show that P. gallinaceum infected A. aegypti presented a reduction in reproductive fitness, accompanied by an activated innate immune response and increase in lipophorin expression, with the latter possibly representing a nutritional resource for Plasmodium sporozoites. (C) 2010 Elsevier Ltd. All rights reserved.

Biologia reprodutiva da |Tainha, Mugil curema valencientes, 1836 (osteichthyes: mugilidade) nas águas costeiras do Rio Grande do Norte

The marine fish white mullet, Mugil curema Valenciennes, 1836 (Osteichthyes: Mugilidae) exhibits a wide geographical distribution, being common in the Brazilian coast and is an important component of the artisanal fisheries. The objective of this study was to investigate the reproductive biology of M. curema in the coastal waters of Rio Grande do Norte. Fish samples were captured on a monthly basis during August, 2008 to July, 2009. The fish specimens were numbered, weighed, measured, dissected and their gonads were removed, weighed, their sex and gonadal development were identified. The length-weight relationship was determined for males and females. The sex ratio, the size at first gonadal maturation and gonadosomatic index (GSI) were calculated and ovarian development was investigated using macroscopic and histological techniques. The fecundity, spawning type and the reproductive period of the species were determined. A total of 366 specimens (186 males and 180 females) were captured. The sex ratio was 1:1 and the females were heavier than males. The estimated values of the angular coefficient for both sexes suggest that the species has isometric growth. The size at which 50% of the population began the process of maturation was 25.9 cm of total length for grouped sex. The macroscopic characteristics of the ovaries showed four stages of development: immature, maturing, mature and spent. However, the microscopic characteristics of the ovaries showed five stages of development: immature, early maturing, late maturing, mature and spent. The development of oocytes indicated five phases: Chromatin-nucleolus (phase I), initial perinucleolar (phase II), final perinuclear (phase II), formation of vitelline vesicle or yolk (phase III), vitellogenic (phase IV) and complete vitellogenesis (phase V). The species has a prolonged spawning period, with two peaks coinciding with the rainy season.

Follicular epithelium, theca and egg envelope formation in Serrasalmus spilopleura (Teleostei, Characiformes, Characidae)

The follicular epithelium and theca of oocytes in Serrasalmus spilopleura differentiates during the initial primary growth phase. The follicular cells are squamous and the thecal cells are disposed in two layers. During the secondary growth phase, follicular cells become cuboidal, acquire characteristics typical of protein- or glycoprotein-producing cells, and show dilated intercellular spaces. Formation of the egg envelope in S. spilopleura begins in the previtellogenic oocytes as a layer of amorphous electron-dense material is laid down on the oolemma. During vitellogenesis, another layer of electron-dense material appears beneath the first layer. Also during this phase, a layer of amorphous, less electron-dense material is formed adjacent to the follicular epithelium. The secondary egg envelope appears at the postvitellogenic phase and is composed of a filamentous and undulant material. The morphology of the egg envelopes in S. spilopleura reflects not only its oviparous nature but also the fact that its eggs are adhesive.

Ultrastructure of the ovary of Dermatobia hominis (Diptera: Cuterebridae): III. Gonial cell degeneration

We studied the ultrastructural aspects of pre-pupae and pupae ovaries of Dermatobia hominis. Physiological degeneration of gonial cells was observed: (a) after the ovarioles differentiation, in the oogonia residing in the apical region of the ovary; (b) at the beginning of vitellogenesis, in the cystoblasts close to the terminal filament. The significance of gonial cell degeneration was correlated with the physiological changes wich occur in the ovary during development.

Ultrastructural study of the follicular epithelium in oocytes of Pachycondyla (Neoponera) villosa ants (Hymenoptera : Ponerinae)

Insect oocytes are surrounded by the follicular epithelium which is simple and cuboidal, wih the mainly functions of: synthesis of vitellin membrane and chorion and synthesis and transport of hemolymph products (proteins). In Pachycondyla (Neoponera) villosa ants workers aged less than 10 days do not present the formation of ovarian follicles (oocytes, nurse cells and follicular cells) indicating that vitellogenesis starts at approximately 10 days of age. Studies of participation of the follicular epithelium in Pachycondyla (Neoponera) villosa showed that in stage I oocytes the epithelium does not present the opening of intercellular spaces. In stage II these spaces begin to be observed together with separation of the follicular epithelium from the oocyte surface. In stage III two types of material were observed in the intercellular spaces: electrodense material in the basal region and compacted material in apical one as well as follicular epithelium/oocytes interface suggesting that the extraovarian material that reach oocytes undergoes some type of modification during passage through the intercellular spaces. The follicular epithelium spaces in queen are bigger than in workers oocytes.

Presence of calcium in oocytes of the diplopod Rhinocricus padbergi Verhoeff (Spirobolida, Rhinocricidae)

In diplopods, the presence of calcium-containing structures seems to be a common finding in some species, with its formation being similar to that observed for other intracellular mineralization systems. In the present study, using histochemistry and transmission electron microscopy, a large amount of calcium was observed in the oocytes of Rhinocricus padbergi. Calcium was detected in both less and well developed oocytes, i.e., the occurrence of calcium coincided with the beginning of vitellogenesis. Calcium was observed as fine granulation distributed within the cytoplasm or deposited in spherical structures apparently formed by overlapping calcium layers. Some authors have suggested that these structures represent a type of reserve used for the calcification of the embryo exoskeleton, whereas others believe that calcium inclusions are a mechanism of organism detoxification as a result of excess calcium ingested by animals during soil turnover. We suggest in this paper that the first hypothesis could be occurring in R. padbergi since at the juvenile stages of the individuals the uptake of calcium is low and because the oocyte is a specialized cell not associated with detoxification.