Abstract
Hypoxia is characterized by low oxygen content in the tissues. The central nervous system (CNS) is highly vulnerable to a lack of oxygen. Prolonged hypoxia leads to the death of brain cells, which underlies the development of many pathological conditions. Despite the relevance of the topic, different approaches used to study the molecular mechanisms of hypoxia have many limitations. One promising lead is the use of various genetically encoded tools that allow for the observation of intracellular parameters in living systems. In the first part of this review, we provide the classification of oxygen/hypoxia reporters as well as describe other genetically encoded reporters for various metabolic and redox parameters that could be implemented in hypoxia studies. In the second part, we discuss the advantages and disadvantages of the primary hypoxia model systems and highlight inspiring examples of research in which these experimental settings were combined with genetically encoded reporters.
Highlights
The evolution of organic life on Earth was accompanied by dramatic changes in the environmental oxygen concentration
Progress in studying hypoxic/ischemic injury of the central nervous system (CNS) only began when the following methods became available: powerful platforms for live imaging that were primarily reliant on optical methods; nuclear technologies such as magnetic resonance imaging (MRI) and positron emission tomography (PET) [16,17], and a plethora of chemogenic and genetically encoded fluorescent indicators for probing a variety of biologically significant ions, signaling molecules, and metabolites, including those involved in redox regulation [18,19,20,21,22]
The ProCY biosensor consists of enhanced cyan fluorescent proteins (FPs) (ECFP) and yellow FP YPet fused by a linker that includes fragments of pVHL (60–154) and hypoxia-inducible factor (HIF)-1α (556–577) separated by 21 amino acid GS-rich sequence (Figure 5D) [123]
Summary
The evolution of organic life on Earth was accompanied by dramatic changes in the environmental oxygen concentration. Progress in studying hypoxic/ischemic injury of the CNS only began when the following methods became available: powerful platforms for live imaging that were primarily reliant on optical methods (fluorescence, confocal and two-photon microscopy, and photoacoustic tomography [15]); nuclear technologies such as magnetic resonance imaging (MRI) and positron emission tomography (PET) [16,17], and a plethora of chemogenic and genetically encoded fluorescent indicators for probing a variety of biologically significant ions, signaling molecules, and metabolites, including those involved in redox regulation [18,19,20,21,22] They are the principal tools used for clinical and diagnostic purposes, nuclear technologies provide insufficient spatiotemporal resolution of brain functioning at a cellular level. Hope that it will encourage future researchers to enter this fruitful scientific field
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