Improved isolation of good-quality total RNA from the optic stalk of mud crab, scylla paramamosain

An improved and efficient protocol was developed based on the TaKaRa RNAiso Plus Kit (Code: D9108A) for isolating good-quality total RNA from the optic stalk of mud crab, Scylla paramamosain. The protocol was based on the Trizol method with modifications. The carapace overlapping the optic stalk was retained with RNA in regular protocol. In order to remove the abundant deposition correlative with the carapace which makes the isolation of RNA particularly difficult, 5M potassium acetate solution (pH = 6.0) was added before the precipitation of RNA, and the temperature of RNA deposition was also decreased to -70ºC to ensure the stabilization of RNA. Good-quality total RNA from the optic stalk of S. paramamosain could be easily isolated with this modified protocol and three conventional methods were also employed to confirm the quality of RNA. This improved method would be helpful in facilitating molecular research of crabs involving RNA from the optic stalk.

Adubação nitrogenada e potássica no desenvolvimento de mudas de maracujazeiro / Nitrogen and potassic fertilization on development of passion fruit seedlings

Este trabalho objetivou avaliar os efeitos da adubação com N e K no desenvolvimento de mudas de maracujazeiro-amarelo (Passiflora edulis) cultivadas em Latossolo Vermelho distrófico. O delineamento experimental foi o inteiramente casualizado, com três repetições, num esquema fatorial 4 x 4, com 4 doses de N (zero, 150, 300 e 600mg de N dm-3) e quatro de K (zero, 75, 150 e 300mg de K dm-3), parceladas em quatro vezes. A unidade experimental foi constituída por vasos de 3,3L. Após 84 dias da germinação, foram avaliados: altura da planta, o diâmetro do caule, o número de folhas e a área foliar. O melhor desenvolvimento das mudas de maracujazeiro ocorreu com as doses de 150mg de N dm-3 e de 300mg de K dm-3.

Preferential Suppression of the On Pathway by r-Aminobutyric Acid in the Catfish Retina

The effects of r-aminobutyric acid(GABA) agonsits and antagonists were explored by the intracellular recording method to discern the preferential suppression of the ON component by GABA on the ON-OFF transient cell in the catfish (Ictalurus punctatus) retina. Experiments were performed in the superfused eyecup preparation. The animals were decapitated and pited before the eye, and the surrounding tissue was removed from the skull. The retina was exposed by excising the cornea, iris, and vitreous. This preparation rested on a wad of Ringer`s soaked cotton in contact with an Ag/Agcl reference electrode. Solutions were delivered through a manifold system that was connected to a pipette located near the absorbent wick. Electro-physiological recordings were made using standard intracellular electrodes filled with 2 M potassium acetate. The electrical signal was recorded with an amplifierand a penwriter, viewed on an oscilloscope, and stored on a data recorder. The light sources were red light-emitting-diode (LED) and the stimuli were full field illumination covering the cntire retina. GABA preferentially reduced ON light responses in ON-OFF transient cell. and GABA hyperpolarized bipolar cells, but the effects on ON bipolar cells were more sensitive than OFF bipolar cells. CACA and TACA, GABAc receptor agonist, did not act on bipolar cells. CACA and TACA, GABAc receptor agonists, hyperpolarized bipolar cells but the sensitivity deferences between ON and OFF bipolar cell were not observed. These results suggest that the preferential suppression of the ON component of the ON-OFF transient cell by GABA was resulted from the presynaptic mechanism that reduced bipolar cell input.

Effects of Extracellular Chloride Ions on the Catfish Retinal Neurons

The catfish (Ictalurus punctatus) retinal neurons were investigated by using the intracellular recording techniques to analyze the function of the chloride ions in the light responses and the ionic mechanisms of the depolarizing actions by GABA. Experiments were performed in the superfused retina-eyecup preparation. The retina was exposed by exicising the cornea, iris, and vitreous. A piece of absorbent tissue with a hole large enough to expose the retina was centered over the eyecup to serve as a wick to draw off the superfusate. Diffuse light stimuli were generated by light-emitting diode positioned above the eyecup. The recordings were made with the use of borosilicate glass micropipettes fashioned from' omega dot' capillary tubing filled with 2 M potassium acetate. Voltage recordings were obtained using an amplifier and amplified signals were recorded on a storage oscillocope, penwriter, and a data recorder. In the catfish retina, the dark membrane potentials were depolarized and the light evoked responses were enhanced in the chloride"-free medium on the catfish horizontal cells. The amplitude of the light evoked potentials were increased by chloride free Ringer's solution on the ON- and OFF-bipolar cells. But the dark membrane potentials were hyperpolarized on the ON-bipolar cell and depolarized on the OFF-bipolar cells in the chloride free medium. The chloride free Ringer's solution changed the light response from ON-sustained to OFF-sustained without any change in amplitude on the ON-sustained cell. The depolarizing actions by GABA on the horizontal cells were maintained in chloride-free environment. But GABA did not abolished the light evoked potentials of the horizontal cell and the ON-sustained cell under the chloride free environment. The results suggest that chloride ion has important roles on the signal transmission of the dark periods in the catfish retina and the depolarizing actions by GABA on the neurons in the catfish retina might be chloride dependent.