The ability of chill-sensitive insects to function at low temperatures limits their geographic ranges. They have species-specific temperatures below which movements become uncoordinated prior to entering a reversible state of neuromuscular paralysis. In spite of decades of research, which in recent years has focused on muscle function, the role of neural mechanisms in determining chill coma is unknown. Spreading depolarization (SD) is a phenomenon that causes a shutdown of neural function in the integrating centres of the central nervous system. We investigated the role of SD in the process of entering chill coma in the locust, Locusta migratoria. We used thermolimit respirometry and electromyography in whole animals and extracellular and intracellular recording techniques in semi-intact preparations to characterize neural events during chilling. We show that chill-induced SD in the central nervous system is the mechanism underlying the critical thermal minimum for coordinated movement in locusts. This finding will be important for understanding how insects adapt and acclimate to changing environmental temperatures.

Temperature has profound effects on the neural function and behaviour of insects. When exposed to low temperature, chill-susceptible insects enter chill coma, a reversible state of neuromuscular paralysis. Despite the popularity of studying the effects of low temperature on insects, we know little about the physiological mechanisms controlling the entry to, and recovery from, chill coma. Spreading depolarization (SD) is a phenomenon that causes a neural shutdown in the central nervous system (CNS) and it is associated with a loss of K + homeostasis in the CNS. Here, we investigated the effects of rapid cold hardening (RCH) on chill tolerance of the migratory locust. With an implanted thermocouple in the thorax, we determined the temperature associated with a loss of responsiveness (i.e. the critical thermal minimum-CT min) in intact male adult locusts. In parallel experiments, we recorded field potential (FP) in the metathoracic ganglion (MTG) of semi-intact preparations to determine the temperature that would induce neural shutdown. We found that SD in the CNS causes a loss of coordinated movement immediately prior to chill coma and RCH reduces the temperature that evokes neural shutdown. Additionally, we investigated a role for octopamine (OA) in the locust chill tolerance and found that OA reduces the CT min and mimics the effects of prior stress (anoxia) in locust.

The ability of chill-sensitive insects to function at low temperatures limits their geographic ranges. They have species-specific temperatures below which movements become uncoordinated prior to entering a reversible state of neuromuscular paralysis. In spite of decades of research, which in recent years has focused on muscle function, the role of neural mechanisms in determining chill coma is unknown. Spreading depolarization (SD) is a phenomenon that causes a shutdown of neural function in the integrating centres of the central nervous system. We investigated the role of SD in the process of entering chill coma in the locust, Locusta migratoria. We used thermolimit respirometry and electromyography in whole animals and extracellular and intracellular recording techniques in semi-intact preparations to characterize neural events during chilling. We show that chill-induced SD in the central nervous system is the mechanism underlying the critical thermal minimum for coordinated movement in locusts. This finding will be important for understanding how insects adapt and acclimate to changing environmental temperatures.

No abstract

Insects live in varied habitats and experience different kinds of environmental stresses. These stresses can impair neural performance, leading to spreading depolarization (SD) of nerve cells and neural shutdown underlying coma. The sensitivity of an insect's nervous system to stress (e.g., anoxia) can be modulated by acute pre-treatment. Rapid cold hardening (RCH) is a form of preconditioning, in which a brief exposure to low temperature can enhance the stress tolerance of insects. SD is associated with a sudden loss of ion, notably K+, homeostasis. We used a pharmacological approach to investigate whether RCH affects anoxia-induced SD in the locust,L. migratoria, via one or more of the following homeostatic mechanisms: (1) Na+/K+-ATPase (NKA), (2) Na+/K+/2Cl-co-transporter (NKCC), and (3) voltage-gated K+(Kv) channels. We also assessed abundance and phosphorylation of NKCC using immunoblotting. We found that inhibition of NKA or Kvchannels delayed the onset of anoxia-induced SD in both control and RCH preparations. However, NKCC inhibition preferentially abrogated the effect of RCH. Additionally, we observed a higher abundance of NKCC in RCH preps but no statistical difference in its phosphorylation level, indicating the involvement of NKCC expression or degradation as part of the RCH mechanism.