Effects of osmotic swelling on voltage-gated calcium channel currents in rat anterior pituitary cells.
ABSTRACT Decrease in extracellular osmolarity ([Os]e) results in stimulation of hormone secretion from pituitary cells. Different mechanisms can account for this stimulation of hormone secretion. In this study we examined the possibility that hyposmolarity directly modulates voltage-gated calcium influx in pituitary cells. The effects of hyposmolarity on L-type (IL) and T-type (IT) calcium currents in pituitary cells were investigated by using two hyposmotic stimuli, moderate (18-22% decrease in [Os]e) and strong (31-32% decrease in [Os]e). Exposure to moderate hyposmotic stimuli resulted in three response types in IL (a decrease, a biphasic effect, and an increase in IL) and in increase in IT. Exposure to strong hyposmotic stimuli resulted only in increases in both IL and IT. Similarly, in intact pituitary cells (perforated patch method), exposure to either moderate or strong hyposmotic stimuli resulted only in increases in both IL and IT. Thus it appears that the main effect of decrease in [Os]e is increase in calcium channel currents. This increase was differential (IL were more sensitive than IT) and voltage independent. In addition, we show that these hyposmotic effects cannot be explained by activation of an anionic conductance or by an increase in cell membrane surface area. In conclusion, this study shows that hyposmotic swelling of pituitary cells can directly modulate voltage-gated calcium influx. This hyposmotic modulation of IL and IT may contribute to the previously reported hyposmotic stimulation of hormone secretion. The mechanisms underlying these hyposmotic effects and their possible physiological relevance are discussed.
- [show abstract] [hide abstract]
ABSTRACT: This study demonstrates that a significant proportion of high voltage-activated (HVA) Ca(2+) influx in native rat anterior pituitary cells is carried through non-L-type Ca(2+) channels. Using whole-cell patch-clamp recordings and specific Ca(+2) channel toxin blockers we show that ~35% of the HVA Ca(2+) influx in somatotrophs and lactotrophs is carried through Ca(v) 2.1, Ca(v) 2.2 and Ca(v) 2.3 channels, and that somatotrophs and lactotrophs share similar proportions of these non-L-type Ca(+2) channels. Furthermore, experiments on mixed populations of native anterior pituitary cells revealed that the fraction of HVA Ca(+2) influx carried through these non-L-type Ca(2+) channels might even be higher (~46%), suggesting that non-L-type channels exist in the majority of native anterior pituitary cells. Using western blotting, immunoblots for α(1C) , α(1D) , α(1A) , α(1B) and α(1E) Ca(+2) channel subunits were identified in native rat anterior pituitary cells. Additionally, using reverse transcriptase (RT)-PCR cDNA transcripts for α(1C) , α(1D) , α(1A) and α(1B) Ca(+2) channel subunits were identified. Transcripts for α(1E) were non-specific and transcripts for α(1S) were not detected at all (control). Altogether these results clearly demonstrate the existence of multiple HVA Ca(+2) channels in the membrane of rat native anterior pituitary cells. Whether these channels are segregated among different membrane compartments was further investigated in flotation assays, demonstrating that Ca(v) 2.1, Ca(v) 1.2 and caveolin-1 were mostly localized in light fractions of Nycodenz gradients, i.e., in lipid raft domains. Ca(v) 1.3 channels were distributed among both light and heavy fractions of the gradients, i.e., among raft and nonraft domains whereas Ca(v) 2.2 and Ca(v) 2.3 channels mostly among nonraft domains. In summary, we demonstrate here multiple pathways for HVA Ca(2+) influx through L-type and non-L-type Ca(2) channels in the membrane of native anterior pituitary cells. Compartmentalization of these channels among raft and nonraft membrane domains might be essential for their proper regulation by separate receptors and signaling pathways. © 2012 The Authors. Journal of Neuroendocrinology © 2012 British Society for Neuroendocrinology.Journal of Neuroendocrinology 08/2012; · 3.33 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: Manganese (Mn(2+))-enhanced magnetic resonance imaging studies of the neuronal pathways of the hypothalamus showed that information about the regulation of food intake and energy balance circulate through specific hypothalamic nuclei. The dehydration-induced anorexia (DIA) model demonstrated to be appropriate for studying the hypothalamus with Mn(2+)-enhanced magnetic resonance imaging. Manganese is involved in the normal functioning of a variety of physiological processes and is associated with enzymes contributing to neurotransmitter synthesis and metabolism. It also induces psychiatric and motor disturbances. The molecular mechanisms by which Mn(2+) produces alterations of the hypothalamic physiological processes are not well understood. (1)H-magnetic resonance spectroscopy measurements of the rodent hypothalamus are challenging due to the distant location of the hypothalamus resulting in limited measurement sensitivity. The present study proposed to investigate the effects of Mn(2+) on the neurochemical profile of the hypothalamus in normal, DIA, and overnight fasted female rats at 14.1 T. Results provide evidence that γ-aminobutyric acid has an essential role in the maintenance of energy homeostasis in the hypothalamus but is not condition specific. On the contrary, glutamine, glutamate, and taurine appear to respond more accurately to Mn(2+) exposure. An increase in glutamine levels could also be a characteristic response of the hypothalamus to DIA.Journal of cerebral blood flow and metabolism: official journal of the International Society of Cerebral Blood Flow and Metabolism 06/2011; 31(12):2324-33. · 5.46 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: Lipid bilayer was deformed by the electrostatic/electrokinetic forces induced by the fixed charges on the top monolayer-solution interface. The strains, stresses and energy were simulated using finite element method. The elastic moduli of the heads were four times greater than those of tails sections, but were individually isotropic. The physics of the situation was evaluated using a coupled system of linear elastic equations and electrostatic-electrokinetic (Poisson-Nernst-Planck) equations. The Coulomb force (due to fixed charges in the electric field), and the dielectric force (due to uneven electric field and the solution-membrane permittivity mismatch) bend the membrane, but unevenly. Whereas the bottom monolayer extends vertically (towards charged surface), the top monolayer compresses. In contrast the top monolayer extends horizontally, but the bottom monolayer compresses. The horizontal normal stress is higher in the heads than in the tails sections, but is similar in two monolayers, whereas the vertical normal stress is small. The horizontal normal stress is associated with horizontal normal strain, and vertical with both vertical and horizontal strain. Surprisingly, the shear stress (an indicator where the membrane will deform), is greater in the tails sections. Finally, the elastic energy (which is clearly greater in the heads sections) is dominated by its horizontal component and peaks in the middle of the membrane. The shear component dominates in the tails sections, and is minimal in the membrane center. Even spatially uniform external force thus leads to complex membrane deformation and generates complex profiles of stress and elastic energy.Biochimica et Biophysica Acta 10/2011; 1818(3):829-38. · 4.66 Impact Factor