Hypoxia Induced Suspended Animation And Recovery In Caenorhabditis Elegance

Abstract

Purpose: Hypoxia induced suspended animation and recovery in Caenorhabditis elegance.

Suspended animation is a state is characterized by the none moving, none feeding and they regain their ability to move and feed and all other functions on recovery from suspended animation. Suspended animation can be induced in invertebrates by exposure to CO2, O2, N2, and H2S and upon stressed condition to young ones at L1 stage. In the present study the worms were exposed in the custom designed Glove box with the supply of Nitrogen N2 till 98.5% of the volume. Inside the glove box worms were recorded using Lumaascope-620 with laptop attached for imaging. Images were taken at 7fps and utilized for the analysis of speed, peed, peristaltic movement, track length, egg hatching and recovery by using N2 gas as an inducer of suspended animation to create an anoxic environment. The time-lapse images were recorded and combined to create the movie for the analysis in WORMLAB software to examine them for movement, speed, track length and peristaltic pumping. Analyses are conducted on the track length of C elegans before, during, and after the period of recovery from suspended animation.

Method: In this study, the worms were exposed in glove box at 99.8% of nitrogen to induce suspended animation and the movement was recorded with the lumascope-620 attached to the microscope on exposure. The revival was assessed by K-medium, M9 and blue light exposure or their effect on suspended animation on worm movement in terms of speed, peristaltic movement, track length, egg hatching and recovery. Time lapsed images were recorded for further analysis by WORMLAB software. Analyses are conducted on the track length of C. elegans before, during and after the period of recovery from suspended animation.

Results: The track length was recorded at 157.328 m/sec prior to hypoxia exposure and 50.389 m/sec during suspended animation. The worms resumed movement and reached a speed of 1177.621 m/sec after three hours of the N2 gas supply being removed or stopped.

Conclusions: The observed speed data indicates that the measured peristaltic track length reduced by 67.98%, while the worm's peristaltic length track increased by 54% following the recovery. The track length recorded was 1227.476m/sec prior to hypoxia exposure it was 74.090m/sec during suspended animation during recovery period the worms resumed movement and reached and the track length was 1083.136 m/sec. The speed as measured by the worm lab based on analysis was a speed of 610.707 m/sec during suspended animation was 74.079 and during recovery period was 1177.621. Similarly peristaltic track lengths before suspended animation was157.328m/sec to hypoxia exposure it was 50.389m/sec during suspended animation and during recovery period it was 242.416 m/sec After three hours of the N2 gas supply being removed or stopped. The observed speed data indicates that the measured speed has been significantly increased during recovery and peristaltic track length reduced by 67.98% during suspended animation, while the worm's peristaltic length track increased by 54% during recovery. The worms reached the state of suspended animation by at 98.5% and above exposure to nitrogen gas and were able to recover after 72h of suspended animation to normalcy.