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L-proline feeding for augmented freeze tolerance of Camponotus japonicus Mayr
Mengjia Dou1, Yazhou Li2, Ziqiao Sun3
1CAS Key Laboratory of Cryogenics, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190, China; School of Engineering Science, University of Chinese Academy of Sciences, Beijing 100049, China; Beijing Key Laboratory of Cryo-Biomedical Engineering, Beijing 100190, China.
Abstract:
The successful cryopreservation of organs is a strong and widespread demand around the world but faces great challenges. The mechanisms of cold tolerance of organisms in nature inspirit researchers to find new solutions for these challenges. Especially, the thermal, mechanical, biological and biophysical changes during the regulation of freezing tolerance process should be studied and coordinated to improve the cryopreservation technique and quality of complex organs. Here the cold tolerance of the Japanese carpenter ants, Camponotus japonicus Mayr, was greatly improved by using optimal protocols and feeding on L-proline-augmented diets for 5 days. When cooling to -27.66 °C, the survival rate of frozen ants increased from 37.50% ± 1.73% to 83.88% ± 3.67%. Profiling of metabolites identified the concentration of whole-body L-proline of ants increased from 1.78 to 4.64 ng g-1 after 5-day feeding. High L-proline level, together with a low rate of osmotically active water and osmotically inactive water facilitated the prevention of cryoinjury. More importantly, gene analysis showed that the expression of ribosome genes was significantly up-regulated and played an important role in manipulating freezing tolerance. To the best of our knowledge, this is the first study to link genetic variation to the enhancement of ants' cold tolerance by feeding exogenous cryoprotective compound. It is worth noting that the findings provide the theoretical and technical foundation for the cryopreservation of more complex tissues, organs, and living organisms.
Insights
Feeding ants L-proline significantly boosted their cold tolerance, increasing survival rates to over 83% at -27.66°C. This study links genetic variation and cryoprotective compounds for potential organ cryopreservation applications.
Area of Science:
- Cryobiology
- Insect Physiology
- Molecular Biology
Background:
- Organ cryopreservation faces significant challenges.
- Understanding natural cold tolerance mechanisms is crucial for developing new cryopreservation techniques.
- Studying thermal, mechanical, biological, and biophysical changes during freezing is key to improving organ cryopreservation.
Purpose of the Study:
- To enhance the cold tolerance of Japanese carpenter ants (Camponotus japonicus Mayr).
- To investigate the role of L-proline and genetic factors in cold tolerance.
- To provide a foundation for cryopreservation of complex tissues and organs.
Main Methods:
- Administering L-proline-augmented diets to ants for 5 days.
- Assessing survival rates after freezing to -27.66°C.
- Profiling whole-body L-proline concentrations.
- Analyzing gene expression, focusing on ribosome genes.
Main Results:
- Survival rate increased from 37.50% to 83.88% after L-proline feeding.
- Whole-body L-proline concentration rose from 1.78 to 4.64 ng/g.
- High L-proline levels and altered water states prevented cryoinjury.
- Ribosome gene expression was significantly up-regulated, contributing to freezing tolerance.
Conclusions:
- L-proline effectively enhances ant cold tolerance and survival.
- Increased L-proline, altered water states, and up-regulated ribosome gene expression are key mechanisms.
- This research offers a novel link between genetic variation and enhanced cold tolerance via exogenous compounds, supporting future organ cryopreservation efforts.
