Model organisms are often used in biological, medical and environmental research. Among insects, Drosophila melanogaster, Galleria mellonella, Apis mellifera, Bombyx mori, Periplaneta americana, and Locusta migratoria are often used. However, new model organisms still appear. In recent years, an increasing number of insect species has been suggested as model organisms in life sciences research due to their worldwide distribution and environmental significance, the possibility of extrapolating research studies to vertebrates and the relatively low cost of rearing. Beetles are the largest insect order, with their representative – Tribolium castaneum – being the first species with a completely sequenced genome, and seem to be emerging as new potential candidates for model organisms in various studies. Apart from T. castaneum, additional species representing various Coleoptera families, such as Nicrophorus vespilloides, Leptinotarsa decemlineata, Coccinella septempunctata, Poecilus cupreus, Tenebrio molitor and many others, have been used. They are increasingly often included in two major research aspects: biomedical and environmental studies. Biomedical studies focus mainly on unraveling mechanisms of basic life processes, such as feeding, neurotransmission or activity of the immune system, as well as on elucidating the mechanism of different diseases (neurodegenerative, cardiovascular, metabolic, or immunological) using beetles as models. Furthermore, pharmacological bioassays for testing novel biologically active substances in beetles have also been developed. It should be emphasized that beetles are a source of compounds with potential antimicrobial and anticancer activity. Environmental-based studies focus mainly on the development and testing of new potential pesticides of both chemical and natural origin. Additionally, beetles are used as food or for their valuable supplements. Different beetle families are also used as bioindicators. Another important research area using beetles as models is behavioral ecology studies, for instance, parental care. In this paper, we review the current knowledge regarding beetles as model organisms and their practical application in various fields of life science.

Temperature stress is one of the crucial factors determining geographical distribution of insect species. Most of them are active in moderate temperatures, however some are capable of surviving in extremely high as well as low temperatures, including freezing. The tolerance of cold stress is a result of various adaptation strategies, among others the mitochondria are an important player. They supply cells with the most prominent energy carrier—ATP, needed for their life processes, but also take part in many other processes like growth, aging, protection against stress injuries or cell death. Under cold stress, the mitochondria activity changes in various manner, partially to minimize the damages caused by the cold stress, partially because of the decline in mitochondrial homeostasis by chill injuries. In the response to low temperature, modifications in mitochondrial gene expression, mtDNA amount or phosphorylation efficiency can be observed. So far study also showed an increase or decrease in mitochondria number, their shape and mitochondrial membrane permeability. Some of the changes are a trigger for apoptosis induced via mitochondrial pathway, that protects the whole organism against chill injuries occurring on the cellular level. In many cases, the observed modifications are not unequivocal and depend strongly on many factors including cold acclimation, duration and severity of cold stress or environmental conditions. In the presented article, we summarize the current knowledge about insect response to cold stress focusing on the role of mitochondria in that process considering differences in results obtained in different experimental conditions, as well as depending on insect species. These differentiated observations clearly indicate that it is still much to explore. Graphical Abstract

Insects are the largest group of animals. They are capable of surviving in virtually all environments from arid deserts to the freezing permafrost of polar regions. This success is due to their great capacity to tolerate a range of environmental stresses, such as low temperature. Cold/freezing stress affects many physiological processes in insects, causing changes in main metabolic pathways, cellular dehydration, loss of neuromuscular function, and imbalance in water and ion homeostasis. The neuroendocrine system and its related signaling mediators, such as neuropeptides and biogenic amines, play central roles in the regulation of the various physiological and behavioral processes of insects and hence can also potentially impact thermal tolerance. In response to cold stress, various chemical signals are released either via direct intercellular contact or systemically. These are signals which regulate osmoregulation – capability peptides (CAPA), inotocin (ITC)-like peptides, ion transport peptide (ITP), diuretic hormones and calcitonin (CAL), substances related to the general response to various stress factors – tachykinin-related peptides (TRPs) or peptides responsible for the mobilization of body reserves. All these processes are potentially important in cold tolerance mechanisms. This review summarizes the current knowledge on the involvement of the neuroendocrine system in the cold stress response and the possible contributions of various signaling molecules in this process.

Protective mechanisms against cold stress are well studied in terrestrial and polar insects; however, little is known about these mechanisms in tropical insects. In our study, we tested if a tropical cockroach Gromphadorhina coquereliana, possesses any protective mechanisms against cold stress. Based on the results of earlier studies, we examined how short-term (3 h) cold (4°C) influences biochemical parameters, mitochondrial respiration activity, and the level of HSPs and aquaporins expression in the fat body and leg muscles of G. coquereliana. Following cold exposure, we found that the level of carbohydrates, lipids and proteins did not change significantly. Nevertheless, we observed significant changes in mitochondrial respiration activity. The oxygen consumption of resting (state 4) and phosphorylating (state 3) mitochondria was altered following cold exposure. The increase in respiratory rate in state 4 respiration was observed in both tissues. In state 3, oxygen consumption by mitochondria in fat body was significantly lower compared to control insects, whereas there were no changes observed for mitochondria in muscle tissue. Moreover, there were cold-induced changes in UCP protein activity, but the changes in activity differed in fat body and in muscles. Additionally, we detected changes in the level of HSP70 and aquaporins expression. Insects treated with cold had significantly higher levels of HSP70 in fat body and muscles. On the other hand, there were lower levels of aquaporins in both tissues following exposure to cold. These results suggest that fat body play an important role in protecting tropical insects from cold stress.

Cold tolerance is considered an important factor determining geographic distribution of insects. We9ve previously shown that despite tropical origin, cockroach Gromphadorinha coquereliana is capable of surviving exposures to cold. However, freezing tolerance of this species had not yet been examined. Low temperature is known to alter membranes integrity in insects but whether chilling or freezing compromises DNA integrity remains a matter of speculation. In the present study, we subjected the G. coquereliana adults to freezing to determine their supercooling point (SCP) and evaluated whether the cockroaches were capable of surviving partial and complete freezing. Next, we conducted single cell gel electrophoresis assay (SCGE) to determine whether heat, cold and freezing altered haemocytes DNA integrity. The SCP of this species was high and around -4.76°C, which is within typical range of freezing-tolerant species. Most cockroaches survived one day after partial ice formation (20% mortality), but died progressively in the next few days after cold stress (70% mortality after 4 days). One day after complete freezing, most insects died (70% mortality), and after 4 days, 90% of them had succumbed. The SCGE assays showed substantial level of DNA damage in haemocytes. When cockroaches were heat-stressed, the level of DNA damage was similar to that observed in the freezing treatment; though all heat-stressed insects survived. The study shows that G. coquereliana can surprisingly be considered as moderately freezing-tolerant species, and for first time that extreme low temperature stress can affect DNA integrity, suggesting that this cockroach may possess an efficient DNA repair system.