Two parallel pathways are required for ultrasound-evoked behavioral changes in Caenorhabditis elegans

Ultrasound has been shown to affect the function of both neurons and non-neuronal cells. However, the underlying molecular machinery has been poorly understood. Here, we show that at least two mechanosensitive proteins act in parallel to generate C. elegans behavioral responses to ultrasound stimuli. We first show that these animals generate reversals in response to a single 10 msec pulse from a 2.25 MHz ultrasound transducer. Next, we show that the pore-forming subunit of the mechanosensitive channel TRP-4, and a DEG/ENaC/ASIC ion channel MEC-4, are both required for this ultrasound-evoked reversal response. Further, the trp-4 mec-4 double mutant shows a stronger behavioral deficit compared to either single mutant. Finally, overexpressing TRP-4 in specific chemosensory neurons can rescue the ultrasound-triggered behavioral deficit in the mec-4 null mutant, suggesting that these two pathways act in parallel. Together, we demonstrate that multiple mechanosensitive proteins likely cooperate to transform ultrasound stimuli into behavioral changes.

Electrofusion of mesophyll protoplasts from two varieties of sugar beet, (Beta vulgaris L.)

Somatic hybridization between different plants through protoplast fusion represent an efficient experimental approach to produce genetically transformed plant species. Electrofution of mesophyll protoplasts in sugar beet was occurred to overcome the barriers faced breeding program of this economically industrial crop Protoplasts were successfully isolated from leave's mesophyll of two varieties of sugar beet (Beta vulgaris L.). Various enzyme solutions were assessed for the cell wall degrading ability. They express different efficiency in isolation of mesophyll protoplasts of var. Baraka. The protoplasts yield was 18 × 104 cell ml-1 using the mixture consisting of 0.5% Cellulase RS, 1.0% Hemicellulase and 0.1% Pectolyase Y-23 with 13% mannitol. A total of 16 hrs. for cell wall digestion, and protoplast viability approached 93%. Protoplasts were isolated from leaf mesophyll of var. Carola using the same enzymatic mixtures. High protoplasts yield 20 × 104 cell ml-1 was obtained, requiring the same period 16 hrs. to approach viability 96%. The protoplasts were spherical in shape, varied in chloroplast distribution, having size ranged 12 – 52 µm. The present study succeeded in electrofusion between Baraka × Carola mesophyll protoplasts, producing somatic hybrid cells under conditions of 1MHz, 1000 Vcm-1, 2 pulses, 1.5 msec./pulse with fusion percent of 73%.

Direct Nerve Implantation Vs. Nerve-Muscle Pedicle: A Comparative Study of Reinnervation in the Rabbit

To determine the optimal method for reinnervation of the paralyzed head and neck musculature, we compared direct muscular nerve implants (DNI) with nerve-muscle pedicles (NMP) in rabbits. In 25 anesthetized animals, one ansa hypoglossi nerve was cut. Five animals sensed as controls and two groups of 10 each received cross-over DNIs or NMP from one sternothyroid to the contralateral sternohyoid muscle. The transplanted nerves of animals that survived long enough for neurotization to occur (8 DNIs, 5 NMPs) were stimulated with 3 to 10 mA. 0.05 msec pulse trains to obtain force curves from corresponding straps. Fiber diameters and areas were calculated on muscles harvested before the animals were killed. There was a nonsignificant trend toward stronger contraction in the NMP group, but NMP fibers were significantly larger than those in DNI and control groups ( p < 0.001).