scholarly journals In vivo fiber photometry of neural activity in response to optogenetically manipulated inputs in freely moving mice

2017 ◽  
Vol 10 (05) ◽  
pp. 1743001 ◽  
Author(s):  
Liang Li ◽  
Yajie Tang ◽  
Leqiang Sun ◽  
Khaista Rahman ◽  
Kai Huang ◽  
...  
Keyword(s):  
Neuronal Activity ◽  
Low Cost ◽  
Functional Study ◽  
V1 Neurons ◽  
Dynamics Of Population ◽  
Lateral Posterior ◽  
Freely Moving ◽  
Final Confirmation ◽  
Detailed Protocol

In vivo fiber photometry is a powerful technique to analyze the dynamics of population neurons during functional study of neuroscience. Here, we introduced a detailed protocol for fiber photometry-based calcium recording in freely moving mice, covering from virus injection, fiber stub insertion, optogenetical stimulation to data procurement and analysis. Furthermore, we applied this protocol to explore neuronal activity of mice lateral-posterior (LP) thalamic nucleus in response to optogenetical stimulation of primary visual cortex (V1) neurons, and explore axon clusters activity of optogenetically evoked V1 neurons. Final confirmation of virus-based protein expression in V1 and precise fiber insertion indicated that the surgery procedure of this protocol is reliable for functional calcium recording. The scripts for data analysis and some tips in our protocol are provided in details. Together, this protocol is simple, low-cost, and effective for neuronal activity detection by fiber photometry, which will help neuroscience researchers to carry out functional and behavioral study in vivo.

scholarly journals sNucDrop-Seq: Dissecting cell-type composition and neuronal activity state in mammalian brains by massively parallel single-nucleus RNA-Seq

2017 ◽  
Author(s):  
Peng Hu ◽  
Emily Fabyanic ◽  
Zhaolan Zhou ◽  
Hao Wu
Keyword(s):  
Single Cell ◽  
Neuronal Activity ◽  
Low Cost ◽  
Single Cells ◽  
High Sensitivity ◽  
Droplet Microfluidics ◽  
Massively Parallel ◽  
Rna Seq ◽  
Single Nucleus

Massively parallel single-cell RNA sequencing can precisely resolve cellular diversity in a high-throughput manner at low cost, but unbiased isolation of intact single cells from complex tissues, such as adult mammalian brains, is challenging. Here, we integrate sucrose-gradient assisted nuclear purification with droplet microfluidics to develop a highly scalable single-nucleus RNA-Seq approach (sNucDrop-Seq), which is free of enzymatic dissociation and nucleus sorting. By profiling ~11,000 nuclei isolated from adult mouse cerebral cortex, we demonstrate that sNucDrop-Seq not only accurately reveals neuronal and non-neuronal subtype composition with high sensitivity, but also enables analysis of long non-coding RNAs and transient states such as neuronal activity-dependent transcription at single-cell resolution in vivo.

scholarly journals GuPPy, a Python toolbox for the analysis of fiber photometry data

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Venus N. Sherathiya ◽  
Michael D. Schaid ◽  
Jillian L. Seiler ◽  
Gabriela C. Lopez ◽  
Talia N. Lerner
Keyword(s):  
Open Source ◽  
User Interfaces ◽  
Low Cost ◽  
Ease Of Use ◽  
Analysis Tool ◽  
Freely Behaving ◽  
Freely Moving ◽  
Computing Platforms ◽  
Graphic User Interfaces

AbstractFiber photometry (FP) is an adaptable method for recording in vivo neural activity in freely behaving animals. It has become a popular tool in neuroscience due to its ease of use, low cost, the ability to combine FP with freely moving behavior, among other advantages. However, analysis of FP data can be challenging for new users, especially those with a limited programming background. Here, we present Guided Photometry Analysis in Python (GuPPy), a free and open-source FP analysis tool. GuPPy is designed to operate across computing platforms and can accept data from a variety of FP data acquisition systems. The program presents users with a set of graphic user interfaces (GUIs) to load data and provide input parameters. Graphs are produced that can be easily exported for integration into scientific figures. As an open-source tool, GuPPy can be modified by users with knowledge of Python to fit their specific needs.

scholarly journals Aversive emotion rapidly activates orexin neurons and increases heart rate in freely moving mice

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Akira Yamashita ◽  
Shunpei Moriya ◽  
Ryusei Nishi ◽  
Jun Kaminosono ◽  
Akihiro Yamanaka ◽  
...  
Keyword(s):  
Heart Rate ◽  
Neuronal Activity ◽  
Defense Response ◽  
Aversive Stimulus ◽  
Critical Role ◽  
Neuron Activity ◽  
Dual Channel ◽  
Arterial Blood ◽  
Freely Moving

AbstractThe perifornical area of the hypothalamus has been known as the center for the defense response, or fight-or-flight response, which is characterized by a concomitant rise in arterial blood pressure, heart rate, and respiratory frequency. It is well established that orexin neurons, which are located in this region, play a critical role in this response. In this study, we further examined this role by recording orexin neuronal activity and heart rate in freely moving mice using an original dual-channel fiber photometry system in vivo. Analysis of orexin neuron activity in relation to autonomic responses to aversive stimuli revealed a rapid increase in neuronal activity just prior to changes in heart rate. In addition, we examined whether orexin neurons would be activated by a conditioned neutral sound that was previously associated with aversive stimulus. We show that the memory of the aversive stimulus activated orexin neurons and increased heart rate. Our data suggest that orexin neurons are a key component linking aversive emotions to autonomic defense response. Our data also suggest that targeting orexin neurons may enable treatment of psychiatric disorders associated with chronic stress and traumatic memories.

scholarly journals Aversive Emotion Activates Orexin Neurons and Induces Tachycardia in Freely Moving Mice

2020 ◽  
Author(s):  
Akira Yamashita ◽  
Shunpei Moriya ◽  
Ryusei Nishi ◽  
Jun Kaminosono ◽  
Akihiro Yamanaka ◽  
...  
Keyword(s):  
Heart Rate ◽  
Neuronal Activity ◽  
Defense Response ◽  
Emotional Memory ◽  
Autonomic Responses ◽  
Dual Channel ◽  
Arterial Blood ◽  
Emotional Component ◽  
Freely Moving

Abstract Stress affects the sensory and negative emotional components in the brain and causes autonomic responses and aversive emotion. However, the possible roles for the negative emotional component remain largely unclear. The perifornical area of the hypothalamus has been known as the center for the defense response, or fight-or-flight response, which is characterized by a concomitant rise in arterial blood pressure, heart rate, and respiratory frequency. Orexin neurons located in that region are suggested to be a critical population responsible for that response. In this study, we examined the suggestion by recording orexin neuronal activity and heart rate in freely moving mice using an original dual-channel fiber photometry system in vivo. Association analysis between orexin neuronal activity and aversive stress-induced autonomic responses revealed a rapid increase in neuronal activity just prior to changes in heart rate. In addition, we examined whether orexin neurons would be activated by a conditioned neutral sound that was previously associated with aversive stimulus. We show that the negative emotional memory indeed activated orexin neurons and increased heart rate. Our data suggest that orexin neurons are the key component required to receive aversive emotion and link it with an autonomic defense response. Our data also suggest that targeting orexin neurons may enable treatment of psychiatric disorders that result from chronic stress and occur long after the original sensory inputs are gone.

scholarly journals GuPPy, a Python toolbox for the analysis of fiber photometry data

2021 ◽  
Author(s):  
Venus N Sherathiya ◽  
Michael D Schaid ◽  
Jillian L Seiler ◽  
Gabriela C Lopez ◽  
Talia Lerner
Keyword(s):  
Open Source ◽  
User Interfaces ◽  
Low Cost ◽  
Ease Of Use ◽  
Analysis Tool ◽  
Development Environment ◽  
Freely Behaving ◽  
Freely Moving ◽  
Graphic User Interfaces

Fiber photometry (FP) is an adaptable method for recording in vivo neural activity in freely behaving animals. It has become a popular tool in neuroscience due to its ease of use, low cost, the ability to combine FP with freely moving behavior, among other advantages. However, analysis of FP data can be a challenge for new users, especially those with a limited programming background. Here, we present Guided Photometry Analysis in Python (GuPPy), a free and open-source FP analysis tool. GuPPy is provided as a Jupyter notebook, a well-commented interactive development environment (IDE) designed to operate across platforms. GuPPy presents the user with a set of graphic user interfaces (GUIs) to load data and provide input parameters. Graphs produced by GuPPy can be exported into various image formats for integration into scientific figures. As an open-source tool, GuPPy can be modified by users with knowledge of Python to fit their specific needs.

A technique for protecting delicate specimens during processing

1989 ◽  
Vol 47 ◽  
pp. 982-983
Author(s):  
R.J. Mount ◽  
R.V. Harrison
Keyword(s):  
Basilar Membrane ◽  
Low Cost ◽  
Organ Of Corti ◽  
Region Of Interest ◽  
Sensory Epithelium ◽  
Physical Damage ◽  
Air Drying ◽  
Specimen Handling ◽  
Two Component

The sensory end organ of the ear, the organ of Corti, rests on a thin basilar membrane which lies between the bone of the central modiolus and the bony wall of the cochlea. In vivo, the organ of Corti is protected by the bony wall which totally surrounds it. In order to examine the sensory epithelium by scanning electron microscopy it is necessary to dissect away the protective bone and expose the region of interest (Fig. 1). This leaves the fragile organ of Corti susceptible to physical damage during subsequent handling. In our laboratory cochlear specimens, after dissection, are routinely prepared by the O-T- O-T-O technique, critical point dried and then lightly sputter coated with gold. This processing involves considerable specimen handling including several hours on a rotator during which the organ of Corti is at risk of being physically damaged. The following procedure uses low cost, readily available materials to hold the specimen during processing ,preventing physical damage while allowing an unhindered exchange of fluids.Following fixation, the cochlea is dehydrated to 70% ethanol then dissected under ethanol to prevent air drying. The holder is prepared by punching a hole in the flexible snap cap of a Wheaton vial with a paper hole punch. A small amount of two component epoxy putty is well mixed then pushed through the hole in the cap. The putty on the inner cap is formed into a “cup” to hold the specimen (Fig. 2), the putty on the outside is smoothed into a “button” to give good attachment even when the cap is flexed during handling (Fig. 3). The cap is submerged in the 70% ethanol, the bone at the base of the cochlea is seated into the cup and the sides of the cup squeezed with forceps to grip it (Fig.4). Several types of epoxy putty have been tried, most are either soluble in ethanol to some degree or do not set in ethanol. The only putty we find successful is “DUROtm MASTERMENDtm Epoxy Extra Strength Ribbon” (Loctite Corp., Cleveland, Ohio), this is a blue and yellow ribbon which is kneaded to form a green putty, it is available at many hardware stores.

Acquired Haemophilia: Functional Study of Antibodies to Factor VIII

1981 ◽  
Vol 45 (03) ◽  
pp. 285-289 ◽  
Author(s):  
J P Allain ◽  
A Gaillandre ◽  
D Frommel
Keyword(s):  
Factor Viii ◽  
Functional Study ◽  
Factor Viii Inhibitor ◽  
Acquired Haemophilia ◽  
Viii Inhibitor ◽  
Kinetics Of ◽  
Antibody Function ◽  
Native Plasma

SummaryFactor VIII complex and its interaction with antibodies to factor VIII have been studied in 17 non-haemophilic patients with factor VIII inhibitor. Low VIII:C and high VIIIR.Ag levels were found in all patients. VIII:WF levels were 50% of those of VTIIRrAg, possibly related to an increase of poorly aggregated and electrophoretically fast moving VIIIR:Ag oligomers.Antibody function has been characterized by kinetics of VIII :C inactivation, saturability by normal plasma and the slope of the affinity curve. Two major patterns were observed:1) Antibodies from 6 patients behaved similarly to those from haemophiliacs by showing second order inhibition kinetics, easy saturability and steep affinity slope (> 1).2) Antibodies from other patients, usually with lower titres, inactivated VIII :C according to complex order kinetics, were not saturable, and had a less steep affinity slope (< 0.7). In native plasma, or after mixing with factor VIII concentrate, antibodies of the second group did not form immune complexes with the whole factor VIII molecular complex. However, dissociation procedures did release some antibodies from apparently low molecular weight complexes formed in vivo or in vitro. For appropriate management of non-haemophilic patients with factor VIII inhibitor, it is important to determine the functional properties of their antibodies to factor VIII.

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