MicroRNA Cues from Nature: A Roadmap to Decipher and Combat Challenges in Human Health and Disease?

MicroRNAs are small non-coding RNA (18–24 nt long) that fine-tune gene expression at the post-transcriptional level. With the advent of “multi-omics” analysis and sequencing approaches, they have now been implicated in every facet of basic molecular networks, including metabolism, homeostasis, and cell survival to aid cellular machinery in adapting to changing environmental cues. Many animals must endure harsh environmental conditions in nature, including cold/freezing temperatures, oxygen limitation (anoxia/hypoxia), and food or water scarcity, often requiring them to revamp their metabolic organization, frequently on a seasonal or life stage basis. MicroRNAs are important regulatory molecules in such processes, just as they are now well-known to be involved in many human responses to stress or disease. The present review outlines the role of miRNAs in natural animal models of environmental stress and adaptation including torpor/hibernation, anoxia/hypoxia tolerance, and freeze tolerance. We also discuss putative medical applications of advances in miRNA biology including organ preservation for transplant, inflammation, ageing, metabolic disorders (e.g., obesity), mitochondrial dysfunction (mitoMirs) as well as specialized miRNA subgroups respective to low temperature (CryomiRs) and low oxygen (OxymiRs). The review also covers differential regulation of conserved and novel miRNAs involved at cell, tissue, and stress specific levels across multiple species and their roles in survival. Ultimately, the species-specific comparison and conserved miRNA responses seen in evolutionarily disparate animal species can help us to understand the complex miRNA network involved in regulating and reorganizing metabolism to achieve diverse outcomes, not just in nature, but in human health and disease.

Freeze tolerance influenced forest cover and hydrology during the Pennsylvanian

The distribution of forest cover alters Earth surface mass and energy exchange and is controlled by physiology, which determines plant environmental limits. Ancient plant physiology, therefore, likely affected vegetation-climate feedbacks. We combine climate modeling and ecosystem-process modeling to simulate arboreal vegetation in the late Paleozoic ice age. Using GENESIS V3 global climate model simulations, varying pCO2, pO2, and ice extent for the Pennsylvanian, and fossil-derived leaf C:N, maximum stomatal conductance, and specific conductivity for several major Carboniferous plant groups, we simulated global ecosystem processes at a 2° resolution with Paleo-BGC. Based on leaf water constraints, Pangaea could have supported widespread arboreal plant growth and forest cover. However, these models do not account for the impacts of freezing on plants. According to our interpretation, freezing would have affected plants in 59% of unglaciated land during peak glacial periods and 73% during interglacials, when more high-latitude land was unglaciated. Comparing forest cover, minimum temperatures, and paleo-locations of Pennsylvanian-aged plant fossils from the Paleobiology Database supports restriction of forest extent due to freezing. Many genera were limited to unglaciated land where temperatures remained above −4 °C. Freeze-intolerance of Pennsylvanian arboreal vegetation had the potential to alter surface runoff, silicate weathering, CO2 levels, and climate forcing. As a bounding case, we assume total plant mortality at −4 °C and estimate that contracting forest cover increased net global surface runoff by up to 6.1%. Repeated freezing likely influenced freeze- and drought-tolerance evolution in lineages like the coniferophytes, which became increasingly dominant in the Permian and early Mesozoic.

Cold Acclimation Increases Freeze Tolerance in Acacia koa, a Tropical Tree Species Occurring over a Wide Elevational Gradient

Frost damage is among the major limitations to reforestation and forest restoration projects worldwide. Investigations of environmental and genetic effects on frost resistance have focused on boreal and temperate tree species rather than tropical trees. Koa (Acacia koa A. Gray) is a valuable tropical hardwood tree species endemic to the Hawaiian Islands, USA. Koa occurs across a wide elevational gradient, and newly planted trees are subject to winter frost at high elevations. We sought to determine whether different koa populations show variation in freeze hardiness as a cold-tolerance mechanism, and whether exposure to hardening conditions prior to frost exposure can modify koa cold-tolerance adaptation. Seeds from 13 populations of koa (Acacia koa A. Gray) were collected across an elevational range (603–2050 m) on the Island of Hawai’i. Four-month-old seedlings grown from the 13 population seed sources were divided into control (non-acclimated) and cold-acclimated treatments, maintained at 26 °C/22 °C (day/night) or exposed to gradually decreasing temperatures to 8 °C/4 °C (day/night), respectively. After six weeks, control and cold-acclimated seedlings from each population were tested for freeze tolerance by electrolyte leakage at five test temperatures ranging from 5 °C (control) to −20 °C. Treatment effects were mainly observed at the lowest test temperatures (−15 and −20 °C). A higher index of cold damage occurred in the non-acclimated seedlings for most of the populations. Several of our higher elevation populations showed greater cold tolerance than populations from lower elevations, particularly when cold-acclimated. Our results suggest that cold acclimation may increase frost hardiness in a tropical forest tree species, and that there is likely some adaptive variation in frost tolerance among populations from different elevations. Cold acclimation could be a useful tool to prepare koa seedlings to be planted in high-elevation sites prone to freezing winter temperatures.

Freeze tolerance in neotropical frogs: an intrageneric comparison using Pristimantis species of high elevation and medium elevation

Paramos are high-elevation tropical Andean ecosystems above the tree line that display variable temperature and frequent freezing spells. Because a significant anuran community lives in this environment, physiological protection against freezing must characterise individuals in this community. Antifreeze protection has been studied in amphibians from other communities, and it is likely that Paramo anurans rely on the same underlying molecules that convey such protection to Nearctic species. However, given the pervasive presence of freezing spells in the Paramos year-round, the processes of activating protection mechanisms may differ from that of seasonal counterparts. Accordingly, this study investigated cryoprotection strategies in high-elevation tropical frogs, using as a model the terrestrial and nocturnal genus Pristimantis, specifically P. bogotensis, P. elegans and P. nervicus from Paramos, and the warm ecosystem counterparts P. insignitus, P. megalops and P. sanctaemartae. We focused on freeze tolerance and its relationship with glucose accumulation and ice formation. Under field conditions, the highest elevation P. nervicus exhibited higher glucose concentration at dawn compared to noon (1.7 ± 0.6 mmol/L versus 3.5 ± 1.32 mmol/L). Under experimental thermal freeze exposure for 2 hours between −2 and −4 ºC, the glucose concentration of the three Paramo species increased but physiological diversity was evident (P. nervicus 126%; P. bogotensis 100%; and P. elegans 55%). During this test, body ice formation was assessed calorimetrically. The species with the highest body ice formation was P. bogotensis (17% ± 5.37; maximum value: 63%; n = 8), followed by P. nervicus (5% ± 3.27; maximum value: 11%; n = 5) and P. elegans (0.34% ± 0.09; maximum value: 1%; n = 4). The study shows physiological diversity both within a genus and across the amphibian community around the freezing contour. Overall, Paramo species differ in freezing physiology from their low-elevation counterparts. Thus, climate shifts increasing freezing spells may affect the structure of communities in this zone.

Comparative Analysis of piRNA Profiles Helps to Elucidate Cryoinjury Between Giant Panda and Boar Sperm During Cryopreservation

Cryopreservation induces sperm cryoinjuries, including physiological and functional changes. However, the molecular mechanisms of sperm cryoinjury and cryoresistance are still unknown. Cryoresistance or the freeze tolerance of sperm varies across species, and boar sperm is more susceptible to cold stress. Contrary to boar sperm, giant panda sperm appears to be strongly freeze-tolerant and is capable of surviving repeated cycles of freeze-thawing. In this study, differentially expressed (DE) PIWI-interacting RNAs (piRNAs) of fresh and frozen-thawed sperm with different freeze tolerance capacity from giant panda and boar were evaluated. The results showed that 1,160 (22 downregulated and 1,138 upregulated) and 384 (110 upregulated and 274 downregulated) DE piRNAs were identified in giant panda and boar sperm, respectively. Gene ontology (GO) enrichment analysis revealed that the target DE messenger RNAs (mRNAs) of DE piRNAs were mainly enriched in biological regulation, cellular, and metabolic processes in giant panda and boar sperm. Moreover, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that the target DE mRNAs of DE piRNAs were only distributed in DNA replication and the cyclic adenosine monophosphate (cAMP) signaling pathway in giant panda, but the cAMP, cyclic guanosine monophosphate (cGMP), and mitogen-activated protein kinase (MAPK) signaling pathways in boar sperm were considered as part of the olfactory transduction pathway. In conclusion, we speculated that the difference in the piRNA profiles and the DE piRNAs involved in the cAMP signaling pathway in boar and giant panda may have contributed to the different freeze tolerance capacities between giant panda and boar sperm, which helps to elucidate the molecular mechanism behind sperm cryoinjury and cryoresistance.

Quantifying Freeze Tolerance of Hybrid Bermudagrasses Adapted for Golf Course Putting Greens

The susceptibility of warm-season turfgrasses such as bermudagrass (Cynodon spp.) to winter injury in the transition zone is a major concern. Therefore, the objective of the study was to evaluate five golf course putting green-type experimental genotypes (OKC6318, OKC0805, OKC1609, OKC0920, and OKC3920) and three commercially available bermudagrasses (‘Champion Dwarf’, ‘TifEagle’, and ‘Tahoma 31’) for freeze tolerance by subjecting them to 11 freezing temperatures (–4 to –14 °C) under controlled environment conditions. The experiment was conducted in batches, with four genotypes per batch, and each batch was replicated in time. The mean lethal temperature to kill 50% of the population (LT50) for each genotype was determined. There were significant differences in LT50 values among the bermudagrass genotypes. ‘Champion Dwarf’ had an LT50 value ranging from –5.2 to –5.9 °C across all three batches. The experimental genotypes tested in this study had LT50 values ranging from –7.0 to –8.1 °C and were each lower than that of ‘Champion Dwarf’. ‘Tahoma 31’, the top performing genotype, had an LT50 value ranging from –7.8 to –9.0 °C across all three batches. OKC 3920 was the only experimental genotype with an LT50 value in the same statistical group as ‘Tahoma 31’. The information gained from this research would be useful for breeders to gauge the genetic gain in freeze tolerance in breeding golf course putting green-type bermudagrass.

Odorant and Taste Receptors in Sperm Chemotaxis and Cryopreservation: Roles and Implications in Sperm Capacitation, Motility and Fertility

Sperm chemotaxis, which guide sperm toward oocyte, is tightly associated with sperm capacitation, motility, and fertility. However, the molecular mechanism of sperm chemotaxis is not known. Reproductive odorant and taste receptors, belong to G-protein-coupled receptors (GPCR) super-family, cause an increase in intracellular Ca2+ concentration which is pre-requisite for sperm capacitation and acrosomal reaction, and result in sperm hyperpolarization and increase motility through activation of Ca2+-dependent Cl¯ channels. Recently, odorant receptors (ORs) in olfactory transduction pathway were thought to be associated with post-thaw sperm motility, freeze tolerance or freezability and cryo-capacitation-like change during cryopreservation. Investigation of the roles of odorant and taste receptors (TRs) is important for our understanding of the freeze tolerance or freezability mechanism and improve the motility and fertility of post-thaw sperm. Here, we reviewed the roles, mode of action, impact of odorant and taste receptors on sperm chemotaxis and post-thaw sperm quality.

Hyperspectral Imaging with Cost-Sensitive Learning for High-Throughput Screening of Loblolly Pine (Pinus taeda L.) Seedlings for Freeze Tolerance

HighlightsA hyperspectral imaging approach was developed for freeze-tolerance phenotyping of loblolly pine seedlings.Image acquisition was conducted before and periodically after artificial freezing of the seedlings.A hyperspectral data processing pipeline was developed to extract the spectra from seedling segments.Cost-sensitive support vector machine (SVM) was used for classifying stressed and healthy seedlings.Post-freeze scanning of seedlings on day 41 achieved the highest screening accuracy of 97%.Abstract. Loblolly pine (Pinus taeda L.) is a commercially important timber species planted across a wide temperature gradient in the southeastern U.S. It is critical to ensure that the planting stock is suitably adapted to the growing environment to achieve high productivity and survival. Long-term field studies, although considered the most reliable method for assessing cold hardiness of loblolly pine, are extremely resource-intensive and time-consuming. The development of a high-throughput screening tool to characterize and classify freeze tolerance among different genetic entries of seedlings will facilitate accurate deployment of highly productive and well-adapted families across the landscape. This study presents a novel approach using hyperspectral imaging to screen loblolly pine seedlings for freeze tolerance. A diverse population of 1549 seedlings raised in a nursery were subjected to an artificial mid-winter freeze using a freeze chamber. A custom-assembled hyperspectral imaging system was used for in-situ scanning of the seedlings before and periodically after the freeze event, followed by visual scoring of the frozen seedlings. A hyperspectral data processing pipeline was developed to segment individual seedlings and extract the spectral data. Examination of the spectral features of the seedlings revealed reductions in chlorophylls and water concentrations in the freeze-susceptible plants. Because the majority of seedlings were freeze-stressed, leading to severe class imbalance in the hyperspectral data, a cost-sensitive learning technique that aims to optimize a class-specific cost matrix in classification schemes was proposed for modeling the imbalanced hyperspectral data, classifying the seedlings into healthy and freeze-stressed phenotypes. Cost optimization was effective for boosting the classification accuracy compared to regular modeling that assigns equal costs to individual classes. Full-spectrum, cost-optimized support vector machine (SVM) models achieved geometric classification accuracies of 75% to 78% before and within 10 days after the freeze event, and up to 96% for seedlings 41 days after the freeze event. The top portions of seedlings were more indicative of freeze events than the middle and bottom portions, leading to better classification accuracies. Further, variable selection enabled significant reductions in wavelengths while achieving even better accuracies of up to 97% than full-spectrum SVM modeling. This study demonstrates that hyperspectral imaging can provide tree breeders with a valuable tool for improved efficiency and objectivity in the characterization and screening of freeze tolerance for loblolly pine. Keywords: Cost-sensitive learning, Freeze tolerance, Hyperspectral imaging, Plant phenotyping, Support vector machine.

Genome analysis of Plectus murrayi, a nematode from continental Antarctica

Plectus murrayi is one of the most common and locally abundant invertebrates of continental Antarctic ecosystems. Because it is readily cultured on artificial medium in the laboratory and highly tolerant to an extremely harsh environment, P. murrayi is emerging as a model organism for understanding the evolutionary origin and maintenance of adaptive responses to multiple environmental stressors, including freezing and desiccation. The de novo assembled genome of P. murrayi contains 225.741 million base pairs and a total of 14,689 predicted genes. Compared to Caenorhabditis elegans, the architectural components of P. murrayi are characterized by a lower number of protein-coding genes, fewer transposable elements, but more exons, than closely related taxa from less harsh environments. We compared the transcriptomes of lab-reared P. murrayi with wild-caught P. murrayi and found genes involved in growth and cellular processing were up-regulated in lab-cultured P. murrayi, while a few genes associated with cellular metabolism and freeze tolerance were expressed at relatively lower levels. Preliminary comparative genomic and transcriptomic analyses suggest that the observed constraints on P. murrayi genome architecture and functional gene expression, including genome decay and intron retention, may be an adaptive response to persisting in a biotically simplified, yet consistently physically harsh environment.