Upconversion-Luminescent Fiber Microchannel Sensors for Temperature Monitoring at High Spatial Resolution in the Brains of Freely Moving Animals.

Brain temperature is a critical factor affecting neural activity and function, whose fluctuations may result in acute life-threatening health complications and chronic neuropathology. To monitor brain temperature, luminescent nanothermometry (LN) based on upconversion nanoparticles (UCNPs) with low autofluorescence has received extensive attention for its advantages in high temperature sensitivity and high response speed. However, most of current the LNs are based on optical imaging, which fails in temperature monitoring in deep brain regions at high spatial resolution. Here, the fiber microchannel sensor (FMS) loaded with UCNPs (UCNP-FMS) is presented for temperature monitoring at high spatial resolution in the deep brains of freely moving animals. The UCNP-FMS is fabricated by incorporating UCNPs in microchannels of optical fibers, whose diameter is ∼50 µm processed by femtosecond laser micromachining for spatially resolved sensing. The UCNPs provide thermal-sensitive upconversion emissions at dual wavelengths for ratiometric temperature sensing, ensuring a detection accuracy of ± 0.3 °C at 37 °C. Superior performances of UCNP-FMS are demonstrated by real-time temperature monitoring in different brain regions of freely moving animals under various conditions such as taking food, undergoing anesthesia/wakefulness, and suffering external temperature changes. Moreover, this study shows the capability of UCNP-FMS in distributed temperature sensing in mammalian brains in vivo.

Plug-and-play fiber-optic sensors based on engineered cells for neurochemical monitoring at high specificity in freely moving animals.

In vivo detection of neurochemicals, including neurotransmitters and neuromodulators, is critical for both understanding brain mechanisms and diagnosing brain diseases. However, few sensors are competent in monitoring neurochemical dynamics in vivo at high specificity. Here, we propose the fiber-optic probes based on engineered cells (FOPECs) for plug-and-play, real-time detection of neurochemicals in freely moving animals. Taking advantages of life-evolved neurochemical receptors as key components, the chemical specificity of FOPECs is unprecedented. We demonstrate the applications of FOPECs in real-time monitoring of neurochemical dynamics under various physiology and pathology conditions. With no requirement of viral infection in advance and no dependence on animal species, FOPECs can be widely adopted in vertebrates, such as mice, rats, rabbits, and chickens. Moreover, FOPECs can be used to monitor drug metabolisms in vivo. We demonstrated the neurochemical monitoring in blood circulation systems in vivo. We expect that FOPECs will benefit not only neuroscience study but also drug discovery.

Background inhibited and speed-loss-free volumetric imaging in vivo based on structured-illumination Fourier light field microscopy.

Benefiting from its advantages in fast volumetric imaging for recording biodynamics, Fourier light field microscopy (FLFM) has a wide range of applications in biomedical research, especially in neuroscience. However, the imaging quality of the FLFM is always deteriorated by both the out-of-focus background and the strong scattering in biological samples. Here we propose a structured-illumination and interleaved-reconstruction based Fourier light field microscopy (SI-FLFM), in which we can filter out the background fluorescence in FLFM without sacrificing imaging speed. We demonstrate the superiority of our SI-FLFM in high-speed, background-inhibited volumetric imaging of various biodynamics in larval zebrafish and mice in vivo. The signal-to-background ratio (SBR) is improved by tens of times. And the volumetric imaging speed can be up to 40 Hz, avoiding artifacts caused by temporal under-sampling in conventional structured illumination microscopy. These suggest that our SI-FLFM is suitable for applications of weak fluorescence signals but high imaging speed requirements.

Effects of amylin on food intake and body weight via sympathetic innervation of the interscapular brown adipose tissue.

Objective: Amylin acts on the lateral dorsal tegmental nucleus (LDT), resulting in anorexic and weight-loss effects and activates thermogenesis in the interscapular brown adipose tissue (IBAT). In addition, it induces neuronal nitric oxide synthase (nNOS) and choline acetyltransferase (ChAT)-mediated feeding. However, the influence of the intact sympathetic nervous system (SNS) in mediating amylin's effects has not been fully characterised. We investigated whether extracellular signal-regulated kinase (ERK), nNOS, and ChAT activities in the LDT are responsible for amylin's anorexigenic effects and whether this requires an intact SNS.Methods: C57BL/6J mice [wild-type (WT), sham, and sympathetic denervation of IBAT] were used. Food consumption, body weight, and distribution of pERK, nNOS, and ChAT positive neurons in the brain were examined following acute and chronic amylin administration.Results: Food intake was significantly decreased in WT and sham animals following acute amylin injection, but not in the denervated mice. Chronic amylin reduced body weight and serum glucose levels after 6 weeks, but increased insulin levels; no changes were observed in the denervated mice. Acute amylin increased the expression of nNOS, ChAT, and uncoupling protein-1 in the IBAT of WT and sham mice, while no changes were observed in the denervated mice and pERK from the above effect.Conclusions: Intact SNS of IBAT influences amylin-induced suppression of food intake and body weight, thus affecting nNOS and ChAT signalling in the LDT and locus coeruleus.

Inhibitory effect of central ghrelin on steroid synthesis affecting reproductive health in female mice.

Ghrelin is a 28-amino acid peptide hormone that regulates ovarian steroid hormone synthesis; however, there is limited evidence regarding the regulation of this pathway by ghrelin in mice ovary. Thus, we aimed to investigate whether central ghrelin action plays a role in murine reproductive health by inhibiting steroid synthesis. Further, we sought to examine the mechanism of central ghrelin action in ovarian steroid hormone synthesis. After the administration of intracerebroventricular ghrelin (1 nmol), we found reduced serum concentrations of oestradiol and progesterone and reduced secretion of follicle-stimulating hormone and luteinising hormone. Although ghrelin reduced 3ß-hydroxysteroid dehydrogenase mRNA and protein levels in the hypothalamus, it did not affect the expression of steroidogenic acute regulatory protein and cytochrome P450 17A1. In the ovary, central ghrelin regulation indirectly inhibited the mRNA and protein levels of steroidogenic acute regulatory protein, cytochrome P450 17A1, and 3ß-hydroxysteroid dehydrogenase. Moreover, no changes were observed in the expression of proliferating cell nuclear antigen and phosphorylation of extracellular signal-regulated kinase. We hypothesised that central ghrelin regulation suppressed serum oestradiol and progesterone levels by indirectly inhibiting the expression of steroidogenic acute regulatory protein, cytochrome P450 17A1, and 3ß-hydroxysteroid dehydrogenase in the ovary. In this regulation, the suppressed secretion of the follicle-stimulating hormone and luteinising hormone in the pituitary by ghrelin could be involved. Furthermore, hypothalamic 3ß-hydroxysteroid dehydrogenase expression is reduced by ghrelin injection.

Response of the expression of oxytocin neurons to ghrelin in female mice.

Ghrelin is an orexigenic agonist that acts directly on neurons in the hypothalamus, controlling appetite and energy balance. Although its role in appetite-associated neurons has been described, the relationship between peripheral ghrelin stimulation and oxytocin expression in the paraventricular nucleus is not fully understood. We evaluated the suppressive function of ghrelin in oxytocin-positive paraventricular nucleus neurons in ovariectomized C57BL/6 mice 2 h after ghrelin injection. The results showed that, in intact mice, peripheral ghrelin stimulation activated estrogen receptor alpha-expressing neurons during the estrous cycle and that agouti-related peptide mRNA expression was remarkably increased. Agouti-related peptide neuron axons co-localized with oxytocin neurons in the paraventricular nucleus. Moreover, the response of oxytocin-positive paraventricular nucleus neurons to ghrelin was suppressed in the proestrus period, while ghrelin decreased the serum concentration of estradiol in the proestrus phase. These data suggest that ghrelin may suppress oxytocin-positive neuron expression via the arcuate nucleus agouti-related peptide circuit, with the possible influence of estradiol in the murine estrous cycle. Unraveling the mechanism of ghrelin-induced oxytocin expression in the hypothalamus paraventricular nucleus broadens the horizon for ghrelin-related appetite research.

Neuroanatomy of melanocortin-4 receptor pathway in the mouse brain.

OBJECTIVE: Melanocortin-4 receptors (MC4Rs) are key regulators of energy homeostasis and adipose deposition in the central nervous system. Considering that MC4R expression regions and function-related research mainly focus on the paraventricular nucleus (PVN), little is known about their distribution throughout the mouse brain, although its messenger RNA distribution has been analyzed in the rat. Therefore, MC4R protein localization in mouse neurons was the focus of this study. METHODS: MC4R protein distribution was assessed in mice through immunofluorescence and Western blotting. RESULTS: MC4R was differentially expressed throughout the arcuate nucleus (ARC), nucleus of the solitary tract (NTS), raphe pallidus (RPa), medial cerebellar nucleus, intermediolateral nucleus, and brainstem. The highest MC4R protein levels were found in the ARC and ventromedial hypothalamic nucleus, while they were significantly lower in the parabrachial nucleus and NTS. The lowest MC4R protein levels were found in the PVN; there was no difference in the protein levels between the area postrema and RPa. CONCLUSIONS: These data provide a basic characterization of MC4R-expressing neurons and protein distribution in the mouse brain and may aid further research on its role in energy homeostasis.

Melanocortin 4 receptor-mediated effects of amylin on thermogenesis and regulation of food intake.

AIMS: Amylin, a pancreatic hormone cosecreted with insulin, exerts important anorexic and weight-loss effects. Melanocortin 4 receptor (MC4R) signalling plays a critical role in energy homeostasis; however, its role on amylin-dependent regulation of food intake and adaptive thermogenesis of interscapular brown adipose tissue (IBAT) are unclear. In this study, we examined the effects of amylin on food intake and thermogenesis on IBAT via the MC4R pathway in mice. MATERIALS AND METHODS: Acute food consumption and thermogenesis in IBAT were measured in male wild-type (WT) and MC4R-deficient mice following intraperitoneal injection of amylin and SHU9119, an MC3R/4R antagonist, to determine the role of the central melanocortin system on the hypothalamus and IBAT. RESULTS: Amylin (50 µg/kg) suppressed feeding and stimulated thermogenesis on IBAT via activation of the MC4R system in mice. Pharmacological blockade of MC4R using SHU9119 (50 µg/kg) attenuated amylin-induced inhibition of feeding and stimulation of thermogenesis in IBAT. No changes were observed when SHU9119 was injected alone. Moreover, amylin significantly increased MC4R expression and c-Fos neuronal signals in the arcuate nucleus and significantly increased acetyl-CoA carboxylase (ACC) phosphorylation in the hypothalamus and IBAT and uncoupling protein-1 (UCP1) expression in the IBAT of WT mice via the MC4R pathway. CONCLUSION: The melanocortin system was involved in amylin-induced suppression of food intake and activation of thermogenesis in both the hypothalamus and IBAT via modulation of ACC phosphorylation and UCP1 expression.