New insights to the molecular phylogenetics and generic assessment in the Rhacophoridae (Amphibia: Anura) based on five nuclear and three mitochondrial genes, with comments on the evolution of reproduction

The phylogenetic relationships among 12 genera of treefrogs (Family, Rhacophoridae), were investigated based on a large sequence data set, including five nuclear (brain-derived neurotrophic factor, proopiomelanocortin, recombination activating gene 1, tyr

Seasonal pattern of reproduction of Hizikia fusiformis (Sargassaceae, Phaeophyta) from Nanao Island, Shantou, China

The maturation pattern of sexual reproduction in Hizikia fusiformis (Harvey) Okamura (Sargassaceae, Phaeaophyta) was examined in 2003 at Yunao Bay, Nanao Island, Shantou, China. Maturation began in mid-April (seawater temperature 19-21 degrees C), reached the peak in mid-May (maturation rate ca. 70%, and seawater temperature 23.5-25 degrees C) and finished in late-June (seawater temperature 27.5-30 degrees C). The Hizikia plants continued to gain the length from the beginning of maturation season to reach a maximum mean length of 34.8 cm in mid-May, after which the mean length was reduced drastically due to the senescence and rupture of the larger plants in size. The major portion of the mature plants belonged to the larger plants between April and May, but to the smaller ones in June. It is suggested that the plant must achieve a critical size before reproductive maturation occurred. There was a positive relationship between the number of receptacles (NR), as well as the reproductive allocation (RA), and the plant size of Hizikia population, with the recorded maximum values of NR and RA being 1220 and 64.3% respectively, for a single plant.

Control of reproduction rhythmicity by environmental and endogenous signals in Ulva pseudocurvata (Chlorophyta)

A field population of Ulva pseudocurvata Koeman et C. Hoek (hereafter termed Ulva) at Sylt Island (North Sea, Germany) exhibited biweekly peaks of gametophytic reproduction during the colder seasons and approximately weekly peaks during summer. The reproductive events lasted 1-5 d and were separated from each other by purely vegetative phases. Under constant conditions in the laboratory, a free-running rhythm was observed with reproductive peaks occurring approximately every 7 d. When artificial moonlight was provided every 4 weeks, fewer reproductive events occurred, and the reproductive rhythm became synchronized to the environmental artificial moonlight rhythm. In the laboratory, apical disks were entirely converted into reproductive tissue after 8 d cultivation, while almost all basal disks stayed vegetative, which prevented the entire loss of the vegetative thallus during reproductive events. Seasonal size reduction of the thallus occurred from late autumn onward and was determined to be controlled by a genuine photoperiodic response, since size reduction could be induced from May onward by experimental short-day (SD) treatment but was prevented in a long-day (LD) or night-break regime (NB). A daily fine-tuning occurred with gamete release early in the morning at the first sign of daylight, following an obligatory dark ("night") period of at least 1 h duration. No release took place if the overnight dark phase was replaced by continuous light. Blue, green, or red light all triggered gamete release after a dark phase at an irradiance of 0.1 mu mol photons . m(-2) .s(-1), while 0.001 mu mol photons . m(-2) . s(-1) was equivalent to a dark control.

Seasonal variation of reproduction rates and body size of Calanus sinicus in the Southern Yellow Sea, China

Annual variations of egg production rate (EPR) and clutch size of Calanus sinicus, as well as body size of females (prosome length and dry weight), were investigated at a series of stations in the Southern Yellow Sea by onboard incubation. Calanus sinicus was spawning in all the 11 cruises investigated, and the annual variation of EPR was bimodal. Monthly average EPR was highest from May to July, respectively, 5.97, 5.36 and 6.30 eggs female(-1) d(-1), then decreased dramatically to only 1.37 eggs female(-1) d(-1) in August and attained the lowest 1.07 eggs female(-1) d(-1) in October. In November, average EPR increased again to 4.31 eggs female(-1) d(-1). Seasonal variation of clutch size was similar to EPR, except that it decreased gradually after August rather than dramatically as did EPR. Prosome length of females was maximum in May and minimum in October, but dry weight was highest in November. Monthly average EPR correlated better with prosome length than dry weight, while clutch size was rather determined by dry weight of females. It is suggested that egg production of C. sinicus was active during two discontinuous periods when both surface and bottom temperature fell into its favorite range (i.e. 10-23degreesC), and different reproductive strategies were adopted in these two reproductive peaks: other than the highest EPR, longer prosome length was also achieved by C. sinicus from May to July, while females in November developed shorter bodies but accumulated more energy for reproduction.

Spring and autumn reproduction of Calanus sinicus in the Yellow Sea

The fecundity, hatching success and naupliar survival of Calanus sinicus were studied in the Yellow Sea during research cruises in April and October 2006, with emphasis on the regulation of reproduction. During both cruises, the egg production rate (EPR) showed large spatial variations (0 to 25.4 eggs female(-1) d(-1)), generally coinciding with the food availability. In April, the abundant phytoplankton and ciliates in the study area supported active reproduction, which would probably initiate the annual population development. In October, females remained immature in the Yellow Sea Cold Water Mass, likely due to the unfavorable environment (poor food and low temperature). However, reproduction and recruitment were high in the neritic region, which may explain the local population recovery in late autumn. Hatching success varied markedly among stations in April (4 to 85%), whereas it was high overall in the neritic region during October (> 90%). Based on the potential recruitment rates, the spring recruitment would be more important for the annual population dynamics, Female gonad maturity, body size and lipid reserve were examined in relation to fecundity. Regression analyses suggested that the reproductive index (defined as the proportion of females with mature gonads) could be an indicator for the EPR of C. sinicus. Among the regulating factors, external food (ciliates and phytoplankton) seems essential for reproduction, whereas inner lipid reserve may mainly serve metabolic needs. Moreover, fecundity is positively related to body size but independent of temperature, which might exert indirect influences on reproduction.

Identification of Differentially Expressed Genes Between Cloned and Zygote-Developing Zebrafish (Danio rerio) Embryos at the Dome Stage Using Suppression Subtractive Hybridization

Comparative analyses of differentially expressed genes between somatic cell nuclear transfer (SCNT) embryos and zygote-developing (ZD) embryos are important for understanding the molecular mechanism underlying the reprogramming processes. Herein, we used the suppression subtractive hybridization approach and from more than 2900 clones identified 96 differentially expressed genes between the SCNT and ZD embryos at the dome stage in zebrafish. We report the first database of differentially expressed genes in zebrafish SCNT embryos. Collectively, our findings demonstrate that zebrafish SCNT embryos undergo significant reprogramming processes during the dome stage. However, most differentially expressed genes are down-regulated in SCNT embryos, indicating failure of reprogramming. Based on Ensembl description and Gene Ontology Consortium annotation, the problems of reprogramming at the dome stage may occur during nuclear remodeling, translation initiation, and regulation of the cell cycle. The importance of regulation from recipient oocytes in cloning should not be underestimated in zebrafish.

Reproductive effects of prenatal exposure to nonylphenol on zebrafish (Danio rerio)

Mature female and male zebrafish were separated and exposed to nonylphenol (NP) at 0.1, 1, 10, 50, 100 and 500 mu g/L, respectively, for 3 weeks. Gonadosomatic index (GSI) in both sexes and vitellogenin (VTG) induction in males was measured as the bioindicators for the impairment to the parents. The results indicated that 50 mu g/L of NP was the non-observed effect concentration (NOEC) for GSI and VTG induction. Afterwards, the 50 mu g/L NP exposed females and males, and the control females and males were cross-wise pair-bred in the control water for one week to examine the reproductive effects. The embryonic cathepsin D (CAT D) activity, eggshell thickness, fecundity, hatching rate and malformation (vertebral column flexure) rate of offspring were determined in the four pair-bred groups. While endpoints remained unchanged in the groups with exposed males, prenatal exposure of females to 50 mu g/L of NP resulted in the impairment of reproduction in groups with exposed females including inhibition of CAT D activity (P < 0.05), decrease of eggshell thickness (by 23.6%) and elevation of malformation rate (P < 0.001). These results suggested NP could induce reproductive damage to zebrafish at NOEC for parents. The results also imply that alterations of CAT D activity and eggshell thickness may be more sensitive biomarkers to indicate the reproductive effects caused by endocrine disrupting chemicals. (c) 2005 Elsevier Inc. All rights is reserved.

Characterization of transgene integration pattern in F4 hGH-transgenic common carp (Cyprinus carpio L.)

The integration pattern and adjacent host sequences of the inserted pMThGH-transgene in the F4 hGH-transgenic common carp were extensively studied. Here we show that each F4 transgenic fish contained about 200 copies of the pMThGH-transgene and the transgenes were integrated into the host genome generally with concatemers in a head-to-tail arrangement at 4-5 insertion sites. By using a method of plasmid rescue, four hundred copies of transgenes from two individuals of F4 transgenic fish, A and B, were recovered and clarified into 6 classes. All classes of recovered transgenes contained either complete or partial pMThGH sequences. The class I, which comprised 83% and 84.5% respectively of the recovered transgene copies from fish A and B, had maintained the original configuration, indicating that most transgenes were faithfully inherited during the four generations of reproduction. The other five classes were different from the original configuration in both molecular weight and restriction map, indicating that a few transgenes had undergone mutation, rearrangement or deletion during integration and germline transmission. In the five types of aberrant transgenes, three flanking sequences of the host genome were analyzed. These sequences were common carp beta-actin gene, common carp DNA sequences homologous to mouse phosphoglycerate kinase-1 and human epidermal keratin 14, respectively.

Reproductive biology of great cormorant (Phalacrocorax carbo sinensis) in the qinghai-tibet plateau

There have been no detailed studies on reproductive biology of the Great Cormorant (Phalacrocorax carbo sinensis) in Qinghai-Tibet Plateau. We conducted such investigations during the breeding seasons of 1999 and 2000 in Qinghai-Lake Bird Isle, China. Great Cormorants began to migrate to Qinghai-Lake for reproduction from the middle of March and left from early October at the end of reproduction. Nesting periods were from early April to mid June and took 50 days. Egg-laying occurred during the three weeks from the end of April to 20 May. Females typically laid an egg every 1-2 days until clutch completion. Mean clutch size in the study area over two years was 3.3 (SE +/- 0.13, N = 68, range 1-5) and most (66.18%) fell within the range 3-4 eggs. Length of eggs averaged 61.01 mm and breadth averaged 34.13 mm. Fresh egg weight averaged 57.34 g (SE +/- 0.36, range 46.0-73.7 g, N = 179). Hatching success was 48.7% and fledging success was 64.9% over two years. Decline of available fish resources in Qinghai-Lake might be one of main causes of lower reproductive success. The causes of chick loss were possibly high altitude, high winds and prolonged rain.

Histone H2A Has a Novel Variant in Fish Oocytes

Histone variants and their modification have significant roles in many cellular processes. In this study, we identified and characterized the histone H2A variant h2af1o in fish and revealed its oocyte-specific expression pattern during oogenesis and embryogenesis. Moreover, posttranslational modification of H2af1o was observed that results from phosphorylation during oocyte maturation. To understand the binding dynamics of the novel core histone variant H2af1o in nucleosomes, we cloned ubiquitous gibel carp h2afx as a conventional histone control and investigated the dynamic exchange difference in chromatin by fluorescence recovery after photobleaching. H2af1o has significantly higher mobility in nucleosomes than ubiquitous H2afx. Compared with ubiquitous H2afx, H2af1o has a tightly binding C-terminal and a weakly binding N-terminal. These data indicate that fish oocytes have a novel H2A variant that destabilizes nucleosomes by protruding its N-terminal tail and stabilizes core particles by contracting its C-terminal tail. Our findings suggest that H2af1o may have intrinsic ability to modify chromatin properties during fish oogenesis, oocyte maturation, and early cleavage.

Cytoplasmic impact on cross-genus cloned fish derived from transgenic common carp (Cyprinus carpio) nuclei and goldfish (Carassius auratus) enucleated eggs

In previous studies of nuclear transplantation, most cloned animals were obtained by intraspecies nuclear transfer and are phenotypically identical to their nuclear donors; furthermore, there was no further report on successful fish cloning since the report of cloned zebrafish. Here we report the production of seven cross-genus cloned fish by transferring nuclei from transgenic common carp into enucleated eggs of goldfish. Nuclear genomes of the cloned fish were exclusively derived from the nuclear donor species, common carp, whereas the mitochondrial DNA from the donor carp gradually disappeared during the development of nuclear transfer (NT) embryos. The somite development process and somite number of nuclear transplants were consistent with the recipient species, goldfish, rather than the nuclear donor species, common carp. This resulted in a long-lasting effect on the vertebral numbers of the cloned fish, which belonged to the range of goldfish. These demonstrate that fish egg cytoplasm not only can support the development driven by transplanted nuclei from a distantly related species at the genus scale but also can modulate development of the nuclear transplants.