Genetically encoded calcium indicators make it possible to track neural activity on a population-wide level. Here we describe a preparation that enables two-photon imaging of neural activity in an essentially intact fly. We present strategies to minimize motion of the brain, both in prep ...
Recent development of genetically encoded calcium indicators (GECIs) allows us to directly monitor neural activities in in vivo preparations from various organisms, enabling the exploration of neural substrates for complex behaviors. As a showcase for such renovated neuroeth ...
Continuing improvements in genetically encoded calcium indicators (GECIs) make imaging an increasingly attractive method to observe neural activity in the Drosophila brain. Two-photon imaging with GECIs allows calcium signals to be monitored in the entire adult fly central bra ...
A method for imaging the synaptic activity of antennal lobe neurons in the Drosophila brain was developed to visualize and study cellular memory traces. Cellular memory traces are defined as any change in the activity of a neuron that is induced by learning, which subsequently alters the proces ...
Many animals are able to detect a plethora of diverse odorants using arrays of odorant receptors located on the olfactory organs. The olfactory information is subsequently encoded and processed by an overlapping, combinatorial activity of neurons forming complex neural circuits in t ...
Imaging of Ca2+ indicators is widely used to record transient intracellular Ca2+ increases associated with bioelectrical activity. The natural bioluminescent Ca2+ sensor aequorin has been historically the first Ca2+ indicator used to address biological questions. Aequorin is g ...
In experimentally amenable organism models, several different physiological techniques have been developed to functionally record the neuronal activity, with the goal to map the neuronal circuitry and elucidate the neural code underlying major neurophysiological functi ...
Neuropeptide Y (NPY) and growth hormone releasing hormone (GHRH) neurons in the hypothalamic arcuate nucleus play an important role in the neuroendocrine control of energy homeostasis and growth hormone secretion. These neuropeptides are synthesized in such small quantities th ...
Methods to define circuit organization in the brain are largely based upon the axonal transport capabilities of neurons. Numerous tracers have been developed since the 1970s that are sequestered by neurons and transported through axons in either the anterograde or retrograde direct ...
Cerebral perfusion, the rate of blood delivery to brain tissue, plays an important role in tissue viability and brain function. The most commonly used magnetic resonance imaging (MRI) method to assess brain perfusion and tissue haemodynamics in clinical investigations is known as dynam ...
In the past decades, physiological research has progressively moved towards less invasive methods that could be used to study freely moving animals. In this chapter we describe the use of digital infrared thermography to detect changes in skin vasoconstriction, body temperature, brown ...
This chapter describes well-established procedures for multiple-labelling immunofluorescence as applied to peripheral neurons. Tissues are fixed with a mixture of formaldehyde and picric acid, and then processed through solvents (ethanol and dimethyl sulfoxide or xylene) ...
This protocol describes a procedure for combining non-radioactive in situ hybridization (ISH) histochemistry with multi-label fluorescence immunohistochemistry (IHC) on rat brain tissue sections. This allows visualization of multiple mRNA and protein targets located w ...
In situ hybridization (ISH) is a useful method to investigate de novo mRNA expression in tissue sections. The high specificity and sensitivity of this technique combined with the great preservation of tissue and cellular morphology conferred by fixatives such as 4% paraformaldehyde ma ...
Total Internal Reflection Fluorescence Microscopy is a powerful application of fluorescence microscopy that allows selective imaging of fluorescent molecules that are either in or close to the plasma membrane of a cell. Thus, it is ideally suited to imaging the trafficking of molecules ...
The number of neurotransmitter receptors on the plasma membrane is regulated by the traffic of �intracellular vesicles. Golgi-derived vesicles provide newly synthesized receptors to the cell surface, whereas clathrin-coated vesicles are the initial vehicles for sequestrat ...
Neurons generate cell-type-specific action potential (AP) patterns as a result of the integration of synaptic inputs received from many other neurons. In neocortical pyramidal neurons, this AP output is not only transmitted to many other postsynaptic neurons, but also back-propaga ...
This chapter summarizes the extracellular recording and juxtacellular labeling method and its application to the characterization of cardiorespiratory brainstem neurons, although this technique could be applied to any neuron. The combination of this method with immunohis ...
Over the last 20 years, neuroscientists have become increasingly interested in two-photon microscopy. One of the reasons for this interest is that two-photon fluorescence excitation allows counterbalancing the deterioration of the optical signals due to light scattering, and th ...
Mitochondria are semi-autonomous organelles, which are central to cellular energy production and signal transduction. Given the tight integration between mitochondrial and cellular physiology, experimental strategies are required to study mitochondrial (dys)func ...
