A glutamatergic basal forebrain to midbrain circuit mediates wakefulness and defensive behavior.

Defensive behavior, a group of responses that evolved due to threatening stimuli, is crucial for animal survival in the natural environment. For defensive measures to be timely and successful, a high arousal state and immediate sleep-to-wakefulness transition are required. Recently, the glutamatergic basal forebrain (BF) has been implicated in sleep-wake regulation; however, the associated physiological functions and underlying neural circuits remain unknown. Here, using in vivo fiber photometry, we found that BF glutamatergic neuron is activated by various threatening stimuli, including predator odor, looming threat, sound, and tail suspension. Optogenetic activation of BF glutamatergic neurons induced a series of context-dependent defensive behaviors in mice, including escape, fleeing, avoidance, and hiding. Similar to the effects of activated BF glutamatergic cell body, photoactivation of BF glutamatergic terminals in the ventral tegmental area (VTA) strongly drove defensive behaviors in mice. Using synchronous electroencephalogram (EEG)/electromyogram (EMG) recording, we showed that photoactivation of the glutamatergic BF-VTA pathway produced an immediate transition from sleep to wakefulness and significantly increased wakefulness. Collectively, our results clearly demonstrated that the glutamatergic BF is a key neural substrate involved in wakefulness and defensive behaviors, and encodes these behaviors through glutamatergic BF-VTA pathway. Overexcitation of the glutamatergic BF-VTA pathway may be implicated in clinical psychiatric diseases characterized by exaggerated defensive responses, such as autism spectrum disorders.

Titanium dioxide-coated core-shell silica microspheres-based solid-phase extraction combined with sheathless capillary electrophoresis-mass spectrometry for analysis of glyphosate, glufosinate and their metabolites in baby foods.

Because of their extremely low amount in complex samples, it is quite challenging to accurate determine residues of phosphorus-containing amino-acid-like herbicides (PAAHs) in food products. Here we develop novel core-shell mesoporous silica (CSMS) microspheres coated by titanium dioxide (CSMS@TiO2) for extraction and enrichment of PAAHs in baby foods. After the dispersive solid phase extraction (d-SPE), sheathless capillary electrophoresis-mass spectrometry (sheathless CE-MS) is utilized to achieve efficient separation and sensitive detection. The synthesized CSMS@TiO2 composites are characterized by various spectroscopic techniques, proving TiO2 is uniformly distributed onto the channel surface of CSMS. The composites have essential features that are favorable for adsorption of the analytes on the material for d-SPE, including uniform diameter (1.0 µm with a shell thickness of 133 nm), large perpendicular mesopores (15.6 nm), high surface area (101.1 m2/g) and large pore volume (0.4 cm3/g). Taking glyphosate, glufosinate and their main metabolites (aminomethylphosphonic acid and 3-methylphosphinicopropionic acid) as analytes, selective and efficient enrichment is achieved by CSMS@TiO2-based d-SPE through the affinity interaction between titanium dioxide and phosphate groups. Sensitive detection of target compounds is achieved with low limits of quantitation (LOQs) between 0.3-1.6 ng/mL and excellent inter/intra-day repeatability. The compounds in nine different commercial baby foods from local markets are analyzed using the proposed method. Good recoveries of 82.3-102.6% are achieved with low RSDs (n = 5) of 2.1-8.3%. Our study indicates that the proposed CSMS@TiO2-based d-SPE combined with sheathless CE-MS is an accurate and reliable approach for sensitive determination of trace-amount PAAHs and their metabolites in complex samples.

The Modulatory Effect of Motor Cortex Astrocytes on Diabetic Neuropathic Pain.

Diabetic neuropathic pain (DNP) is a common complication of diabetes characterized by persistent pain. Emerging evidence links astrocytes to mechanical nociceptive processing, and the motor cortex (MCx) is a cerebral cortex region that is known to play a key role in pain regulation. However, the association between MCx astrocytes and DNP pathogenesis remains largely unexplored. Here, we studied this association using designer receptors exclusively activated by designer drugs to specifically manipulate MCx astrocytes. We proved that the selective inhibition of MCx astrocytes reduced DNP in streptozocin (STZ)-induced DNP models and discovered a potential mechanism by which astrocytes release cytokines, including TNF-α and IL-1ß, to increase neuronal activation in the MCx, thereby regulating pain. Together, these results demonstrate a pivotal role for MCx astrocytes in DNP pathogenesis and provide new insight into DNP treatment strategies.

Basal forebrain GABAergic neurons promote arousal and predatory hunting.

Predatory hunting is an important approach for animals to obtain valuable nutrition and energy, which critically depends on heightened arousal. Yet the neural substrates underlying predatory hunting remain largely undefined. Here, we report that basal forebrain (BF) GABAergic neurons play an important role in regulating predatory hunting. Our results showed that BF GABAergic neurons were activated during the prey (cricket)-hunting and food feeding in mice. Optogenetic activation of BF GABAergic neurons evoked immediate predatory-like actions to both artificial and natural preys, significantly reducing the attack latency while increasing the attack probability and the number of killed natural prey (crickets). Similar to the effect of activating the soma of BF GABAergic neurons, photoactivation of their terminals in the ventral tegmental area (VTA) also strongly promotes predatory hunting. Moreover, photoactivation of GABAergic BF - VTA pathway significantly increases the intake of various food in mice. By synchronous recording of electroencephalogram and electromyogram, we showed that photoactivation of GABAergic BF - VTA pathway induces instant arousal and maintains long-term wakefulness. In summary, our results clearly demonstrated that the GABAergic BF is a key neural substrate for predatory hunting, and promotes this behavior through GABAergic BF - VTA pathway.

Glutamatergic lateral hypothalamus promotes defensive behaviors.

The glutamatergic lateral hypothalamus (LH) has been implicated in a variety of behaviors, such as evasion and feeding, while its role in defensive behaviors and relevant neurocircuits remains unclear. Here, we demonstrated that the glutamatergic LH is a critical structure regulating defensive behaviors. Trimethylthiazole (TMT), the odor of mice predator, significantly increased c-Fos expression in the LH. Using fiber photometry technology, we found that TMT exposure increased the activity of LH glutamatergic neurons. Selective activation of LH glutamatergic neurons with optogenetics and chemogenetics promoted a series of defense-related behaviors, including fleeing, avoidance, and hiding, while selective inhibition of LH glutamatergic neurons suppressed the avoidance provoked by TMT. Activation of both the glutamatergic LH terminals in the hypothalamic paraventricular nucleus (PVN) and the glutamatergic projection from the basolateral amygdala (BLA) to the LH elicited defensive behaviors. Finally, by combining the viral-mediated retrograde tracing with anterograde activation, we found that PVN-projecting glutamatergic neurons in the LH were activated by BLA glutamatergic inputs. Taken together, our results illustrate that the glutamatergic LH is a pivotal relay of defensive behaviors and possibly promotes these behaviors through the BLA→LH→PVN pathway.

Ventral tegmental area GABAergic neurons induce anxiety-like behaviors and promote palatable food intake.

The obesity epidemic is a global problem and a great challenge for public health. Overconsumption of food, especially palatable food, is the leading cause of obesity. The precise neural circuits underlying food overconsumption remain unclear and require further characterization. In the present study, we showed that Ca2+ signals of GABAergic neurons within the ventral tegmental area (VTA) increased after the onset of food intake, especially high-fat or high-sugar chow. Optogenetic activation of VTA GABAergic neurons evoked immediate eating of palatable food and significantly increased palatable food intake in satiated mice. Photoinhibition of VTA GABAergic neurons suppressed palatable food intake. Surprisingly, photoactivation of VTA GABAergic neurons suppressed the intake of standard chow in fasted mice, but did not reduce the duration of eating of standard chow. Moreover, we found that photoactivation of these neurons drove a series of anxiety-like behaviors in the open field, elevated plus maze, and marble-burying test. Additionally, we found that VTA GABAergic neurons sent abundant projections to the lateral hypothalamus and photoactivation of GABAergic VTA terminals in the lateral hypothalamus induced overconsumption of palatable food, but not anxiety-like behaviors. Taken together, our results illustrate that GABAergic VTA neurons are a key node in the neural circuitry underlying anxiety-like behavior and over-feeding of palatable food, and that over-excitation of GABAergic VTA neurons may underlie clinical diseases related to anxiety and obesity.

Ultrasensitive determination of organotin compounds in plastic food packaging and edible oils by sheathless capillary electrophoresis-electrospray ionization-mass spectrometry.

The determination of trace-amount organotins in plastic food packaging materials is of great significance in food safety. However, due to the complexity of organotins and sample treatment processes, it is still a challenging task. Here, we report a method for the sensitive and simultaneous determination of organotins in plastic food packaging materials and edible oils, by utilizing sheathless capillary electrophoresis-electrospray ionization-mass spectrometry. The method of sample pretreatment with ultrasonic extraction and solid phase extraction is used to eliminate interference. The results showed low limits of detection (LODs) of 2 pg mL-1-50 pg mL-1 and excellent inter/intra-day repeatability. Good average recoveries in the range of 80.27% to 108.52% were obtained at three spiked concentrations, with a relative standard deviation less than 8.71%. The successful simultaneous determination of the target analytes will pave the way for further assessment of contamination and migration behaviour of organotins from packaging materials to food, which is of great significance for evaluating and controlling food safety.

Sensitive detection of benzophenone-type ultraviolet filters in plastic food packaging materials by sheathless capillary electrophoresis-electrospray ionization-tandem mass spectrometry.

While they are commonly used as ultraviolet (UV) filters in plastic food packaging materials, benzophenones (BPs) are reported as environmental endocrine disruptors, and some of them possess significant estrogenic activity. Therefore sensitive determination of the content of those UV filters in plastic polymers is of vital importance in safety assessment of food packaging materials. Here, the sheathless capillary electrophoresis-electrospray ionization-tandem mass spectrometry (CE-ESI-MS/MS) method is applied for the first time to sensitively detect BP-type UV filters in plastic food packaging materials. We investigate and optimize a variety of factors that may affect ESI-MS efficiency and CE separation. Sensitive detection of six BP-type UV filters is achieved using sheathless CE-ESI-MS/MS in conjunction with accelerated solvent extraction and solid phase extraction, with the limit-of-detection of 7 pg/mL-2.4 ng/mL. The method exhibits excellent inter/intra-day repeatability along with the advantages of efficient separation, rapid analysis, low sample consumption and high sensitivity. Six BP-type UV filters in eight different brands of plastic films obtained from supermarkets are successfully analyzed using the method. Good recoveries of 81.3-104.1% at three levels of spiked concentrations are achieved with low RSDs (n = 5) of 2.5-8.7%. Our study shows that the sheathless CE-ESI-MS/MS is a robust and reliable method for sensitive and rapid analysis of UV filters, which would be of potential application in safety assessment of plastic food packaging materials.