Intermittent Binge-Intake Model in Mice

Obesity affects one in eight individuals globally. Overeating, especially high-calorie foods, plays a significant role in obesity. Binge eating disorder ...

Universidad Nacional Autónoma de México (UNAM)

Phosphatidylinositol-4,5-bisphosphate (PIP) is a membrane phosphoinositide that regulates the activity of many ion channels. Influx of calcium primarily through voltage-gated calcium (Ca) channels promotes insulin secretion in pancreatic β-cells. However, whether Ca channels are regulated by PIP, as is the case for some non-insulin-secreting cells, is unknown. The purpose of this study was to investigate whether Ca channels are regulated by PIP depletion in pancreatic β-cells through activation of a muscarinic pathway induced by oxotremorine methiodide (Oxo-M). Ca channel currents were recorded by the patch-clamp technique. The Ca current amplitude was reduced by activation of the muscarinic receptor 1 (MR) in the absence of kinetic changes. The Oxo-M-induced inhibition exhibited the hallmarks of voltage-independent regulation and did not involve PKC activation. A small fraction of the Oxo-M-induced Ca inhibition was diminished by a high concentration of Ca chelator, whereas ≥50% of this inhibition was prevented by diC8-PIP dialysis. Localization of PIP in the plasma membrane was examined by transfecting INS-1 cells with PH-PLCδ1, which revealed a close temporal association between PIP hydrolysis and Ca channel inhibition. Furthermore, the depletion of PIP by a voltage-sensitive phosphatase reduced Ca currents in a way similar to that observed following MR activation. These results indicate that activation of the MR pathway inhibits the Ca channel via PIP depletion by a Ca-dependent mechanism in pancreatic β- and INS-1 cells and thereby support the hypothesis that membrane phospholipids regulate ion channel activity by interacting with ion channels.

PIP2 in pancreatic β-cells regulates voltage-gated calcium channels by a voltage-independent pathway.

The arcuate nucleus (ARC), located at the base of hypothalamus, contains two main populations of neurons involved in the regulation of food intake and energy expenditure. The NPY neurons are orexigenic and their activation stimulates food intake while the activation of POMC neurons promote the opposite effect. Several works have tried to identify these neurons based on their electrophysiological and pharmacological characteristics. However, the classification of ARC neurons is still inconclusive. In this work, glucose concentrations were changed within at physiological range, and the response of rat ARC neurons to this stimulus was used to identify them. Subsequently, the cells were classified on the basis of their passive and active electrophysiological properties. Finally, calcium imaging experiments were done to study the response of ARC neurons populations changing glucose concentrations. We found that NPY and putative POMC neurons can be distinguished based on their electrophysiological properties such as input resistance and firing pattern. Calcium imaging experiments confirmed the diversity of ARC neurons.

Electrophysiological characterization of glucose sensing neurons in the hypothalamic arcuate nucleus of male rats.

There is increasing evidence that the brain resides in a state of criticality. The purpose of the present work is to characterize the dynamics of individual hippocampal CA1 pyramidal cells and to investigate how it is influenced by changes in Kv7.2/7.3 (M-channel) ion channel modulation, which is known to be key in determining the neuronal excitability. We show that the resting activity of CA1 neurons exhibit random dynamics with low information content, while changes in M-channel modulation move the neuronal activity near a phase transition to richer non-trivial dynamics. We interpret these results as the basis upon which the state of self-organized criticality is built.

Dynamical phase transition in spike neuronal firing patterns of hippocampal cells.

Stereotaxic surgery is a less invasive form of surgery that uses a three-dimensional coordinate system to place instruments at a specific location in the brain. Through this type of surgery, one can place needles among other tools within the structures of the brain. Therefore, injections can be given in order to deliver substances that cannot cross the blood-brain barrier. The two most important parameters of the microinjection to control are volume and speed. The volume should not be so large that it displaces the brain tissue and tears it. The injection speed must also be slow so that the liquid that comes out of the syringe can diffuse into the tissue without displacing it and damaging it. Thus, the objectives of the present work are: 1) To develop not a 3D printed prototype but an end-user device. 2) The device must be for animal research only. 3) It must have the same precision in volume and speed as commercial devices. 4) It must be adjustable for microsyringes from 0.5 µl to 1 ml. 5) It must be possible to place it directly on the stereotaxic surgery apparatus and to use it separately. 6) The price must be substantially lower than that of the commercial devices.

Arduino based intra-cerebral microinjector device for neuroscience research.

The plasma membrane Ca-ATPase (PMCA) is crucial for the fine tuning of intracellular calcium levels in eukaryotic cells. In this study, we show the presence of CARC sequences in all human and rat PMCA isoforms and we performed further analysis by molecular dynamics simulations. This analysis focuses on PMCA1, containing three CARC motifs, and PMCA4, with four CARC domains. In PMCA1, two CARC motifs reside within transmembrane domains, while the third is situated at the intracellular interface. The simulations depict more stable RMSD values and lower RMSF fluctuations in the presence of cholesterol, emphasizing its potential stabilizing effect. In PMCA4, a distinct dynamic was found. Notably, the total energy differences between simulations with cholesterol and phospholipids are pronounced in PMCA4 compared to PMCA1. RMSD values for PMCA4 indicate a more energetically favorable conformation in the presence of cholesterol, suggesting a robust interaction between CARCs and this lipid in the membranes. Furthermore, RMSF analysis for CARCs in both PMCA isoforms exhibit lower values in the presence of cholesterol compared to POPC alone. The analysis of H-bond occupancy and total energy values strongly suggests the potential interaction of CARCs with cholesterol. Given the crucial role of PMCAs in physiological calcium regulation and their involvement in diverse pathological processes, this study underscores the significance of CARC motifs and their interaction with cholesterol in elucidating PMCA function. These insights into the energetic preferences associated with CARC-cholesterol interactions offer valuable implications for understanding PMCA function in maintaining calcium homeostasis and addressing potential associated pathologies.

Article	Year
PIP2 in pancreatic β-cells regulates voltage-gated calcium channels by a voltage-independent pathway. American journal of physiology. Cell physiology\| PubMed ID: 27488666	2016
Electrophysiological characterization of glucose sensing neurons in the hypothalamic arcuate nucleus of male rats. Neuroscience letters\| PubMed ID: 30926375	2019
Dynamical phase transition in spike neuronal firing patterns of hippocampal cells. Biochemical and biophysical research communications\| PubMed ID: 31296383	2019
Arduino based intra-cerebral microinjector device for neuroscience research. HardwareX\| PubMed ID: 37457306	2023
Analysis of cholesterol-recognition motifs of the plasma membrane Ca-ATPase. Journal of bioenergetics and biomembranes\| PubMed ID: 38436904	2024

Jorge Bravo-Martinez

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