Maturation of rhesus monkey oocytes in chemically defined culture media and their functional assessment by IVF and embryo development

This study compared success of in-vitro maturation of rhesus monkey oocytes in protein-free versus serum-containing culture systems, assessed by embryo development subsequent to IVF. Four media were tested: (i) modified Connaught Medical Research Laborato

Larval and postlarval development of the window-pane shell, Placuna placenta L. (Bivalvia: Placunidae)

Stages of development of P. placenta from the straight-hinge veliger to the adult are described. Mature larvae metamorphose at lengths from 220-230 m. Larvae probably attach byssally to the water surface at metamorphosis and remain in the plankton for some time before finally settling on the mud bottom.

Captive breeding and embryonic development of honey gourami, Colisa sota (Ham.-Buch.)

Honey Gourami, Colisa sota, has high ornamental as well as food value. The natural resources of this species are gradually declining, due to destruction of its habitat, over fishing for aquarium trade and human consumption. The fish was bred in captivity under controlled environment. It laid about 200-400 eggs in bubble nest built by the male. Hatching started within 28-30hrs. after egg laying. The hatchlings became free swimming by 3rd to 4th day of hatching. The male showed territoriality and parental care by guarding the eggs and hatchlings. The larval survival was 30-35%. The breeding behavior, embryonic and post embryonic development of the fish were studied.

Embryonic and larval development of Mystus gulio (Ham.)

Mystus gulio eggs are strongly adhesive and contain relatively small yolk (0.75-1.0 mm). The egg envelop is thick and transparent. First cleavage (two cells), four cells, eight cells, sixteen cells and multi cells stages were found 20, 25, 35-40, 60 and 70 minutes after fertilization, respectively. The morula stage was visualized within 1.5 h after fertilization. The heart beat visible and the circulatory system commenced after 16 h of fertilization. Embryos hatched 18-20h after activation of egg. The newly hatched larva measured 2.82±0.03 mm in length and 0.32±0.06 mg in weight. The yolk sac was fully absorbed by the third day though larvae commenced exogenous feeding even before completion of yolk absorption. A 5-day old post larva began wandering in search of food. Ten-day old post larvae endowed with eight branched rays in dorsal fin and seven in caudal fin. Fifteen-day old post larvae had the pectm:al spine become stout though the embryonic fin folds had to be disappeared. The length of fingerlings ranged from 25-30 mm after 30 days, and their external features were just like those of an adult except that they were not sexually matured.

Biofluid behaviour in 3D microchannel systems: Numerical analysis and design development of 3D microchannel biochip separators

This paper describes the design and development cycle of a 3D biochip separator and the modelling analysis of flow behaviour in the biochip microchannel features. The focus is on identifying the difference between 2D and 3D implementations as well as developing basic forms of 3D microfluidic separators. Five variants, based around the device are proposed and analysed. These include three variations of the branch channels (circular, rectangular, disc) and two variations of the main channel (solid and concentric). Ignoring the initial transient behaviour and assuming steady state flow has been established, the efficiencies of the flow between the main and side channels for the different designs are analysed and compared with regard to relevant biomicrofluidic laws or effects (bifurcation law, Fahraeus effect, cell-free phenomenon, bending channel effect and laminar flow behaviour). The modelling results identify flow features in microchannels, a constriction and bifurcations and show detailed differences in flow fields between the various designs. The manufacturing process using injection moulding for the initial base case design is also presented and discussed. The work reported here is supported as part of the UK funded 3D-MINTEGRATION project. © 2010 IEEE.

Cooperation of Mtmr8 with PI3K Regulates Actin Filament Modeling and Muscle Development in Zebrafish

Background: It has been shown that mutations in at least four myotubularin family genes (MTM1, MTMR1, 2 and 13) are causative for human neuromuscular disorders. However, the pathway and regulative mechanism remain unknown. Methodology/Principal Findings: Here, we reported a new role for Mtmr8 in neuromuscular development of zebrafish. Firstly, we cloned and characterized zebrafish Mtmr8, and revealed the expression pattern predominantly in the eye field and somites during early somitogenesis. Using morpholino knockdown, then, we observed that loss-of-function of Mtmr8 led to defects in somitogenesis. Subsequently, the possible underlying mechanism and signal pathway were examined. We first checked the Akt phosphorylation, and observed an increase of Akt phosphorylation in the morphant embryos. Furthermore, we studied the PH/G domain function within Mtmr8. Although the PH/G domain deletion by itself did not result in embryonic defect, addition of PI3K inhibitor LY294002 did give a defective phenotype in the PH/G deletion morphants, indicating that the PH/G domain was essential for Mtmr8's function. Moreover, we investigated the cooperation of Mtmr8 with PI3K in actin filament modeling and muscle development, and found that both Mtmr8-MO1 and Mtmr8-MO2+LY294002 led to the disorganization of the actin cytoskeleton. In addition, we revealed a possible participation of Mtmr8 in the Hedgehog pathway, and cell transplantation experiments showed that Mtmr8 worked in a non-cell autonomous manner in actin modeling. Conclusion/Significance: The above data indicate that a conserved functional cooperation of Mtmr8 with PI3K regulates actin filament modeling and muscle development in zebrafish, and reveal a possible participation of Mtmr8 in the Hedgehog pathway. Therefore, this work provides a new clue to study the physiological function of MTM family members.

Antisense for gonadotropin-releasing hormone reduces gonadotropin synthesis and gonadal development in transgenic common carp (Cyprinus carpio)

Generating transgenic fish with desirable traits (e.g., rapid growth, larger size, etc.) for commercial use has been hampered by concerns for biosafety and competition if these fish are released into the environment. These obstacles may be overcome by producing transgenic fish that are sterile, possibly by inhibiting hormones related to reproduction. In vertebrates, synthesis and release of gonadotropin (GtH) and other reproductive hormones is mediated by gonadotropin-releasing hormone (GnRH). Recently two cDNA sequences encoding salmon-type GnRH (sGnRH) decapeptides were cloned from common carp (Cyprinus carpio). This study analyzed the expression of these two genes using real-time polymerase chain reaction (RT-PCR) in different tissues carp at varying developmental stages. Transcripts of both genes were detected in ovary and testis in mature and regressed, but not in juvenile carp. To evaluate the effects of sGnRH inhibition, the recombinant gene CAsGnRHpc-antisense, expressing antisense sGnRH RNA driven by a carp beta-actin promoter, was constructed. Blocking sGnRH expression using antisense sGnRH significantly decreased GtH in the blood of male transgenic carp. Furthermore, some antisense transgenic fish had no gonadal development and were completely sterile. These data demonstrate that sGnRH is important for GtH synthesis and development of reproductive organs in carp. Also, the antisense sGnRH strategy may prove effective in generating sterile transgenic fish, eliminating environmental concerns these fish may raise. (c) 2007 Published by Elsevier B.V.