Volume changes in single N1E-115 neuroblastoma cells measured with a fluorescent probe

Crowe, William E.; Altamirano, Julio; Huerto, L.; Álvarez-Leefmans, Francisco J.

doi:10.1016/0306-4522(95)00219-9

Cited by 111 publications

(116 citation statements)

References 31 publications

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“…We used the intracellularly trapped dye, calcein, measured within a defined intracellular volume (set by the properties of the confocal microscope and the ROI) as a reflection of cytoplasmic volume changes. Calcein is insensitive to changes in intracellular calcium and pH (26) and is only minimally affected by variations in ionic strength (27). Swelling or shrinkage of the cell will be accompanied by a decrease or an increase, respectively, in the dye concentration in the cell (28).…”

Section: Discussionmentioning

confidence: 99%

Changes of Cell Volume and Nitric Oxide Concentration in Macula Densa Cells Caused by Changes in Luminal NaCl Concentration

Liu

Pittner

Persson

2002

Journal of the American Society of Nephrology

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Abstract. The luminal NaCl concentration ([NaCl]) at the macula densa (MD) controls both tubuloglomerular feedback (TGF) and renin release. Nitric oxide (NO) inhibits TGF sensitivity to a great extent. The NO concentration in the MD cells is not known. This study measured this concentration in MD cells with confocal microscopy in the isolated perfused thick ascending limb using a NO-sensitive fluorophore 4,5-diaminofluorescein (DAF-2). Calcein was used to measure cell volume changes. The loop perfusion fluid was a modified Ringer solution containing 10, 35, or 135 mM NaCl with a constant total osmolarity (290 mOsm), and the bath was perfused with the 135 mM NaCl solution. The results show that MD cell volume and NO concentration measured with DAF-2 DA increased considerably with increasing luminal[NaCl] and with calcium-free solutions in the lumen and bath. L-arginine (5 mM) increased NO concentration in the MD cells by 30%. 7-nitroindazole could totally inhibit the NO production caused by L-arginine and by increased luminal [NaCl]. In conclusion, the MD cell volume changes caused by the changes of luminal [NaCl] were quantitatively measured, and it was found that increasing the luminal [NaCl] resulted in an increase in cell volume. It was also found that NO formation in MD cells could be measured with DAF-2 and that NO production was increased through neuronal NO synthase activation with an increased luminal [NaCl]. An increased NO production will inhibit the vasoconstriction induced by the TGF and at the same time will reduce TGF sensitivity.The juxtaglomerular apparatus (JGA) is a complex assembly of specialized structures related to each other anatomically, forming the vascular pole of the glomerulus. The JGA is comprised of the macula densa (MD), the extraglomerular mesangium, and the afferent and efferent arterioles. The MD is a plaque of 20 to 30 specialized epithelial cells belonging to the end portion of the thick ascending limb. The luminal NaCl concentration ([NaCl]) at the MD has two established effects: (1) regulation of glomerular arteriolar resistance through tubuloglomerular feedback (TGF); and (2) control of renin release (1,2). The first step in these signal transmissions involves NaCl transport by the MD, which is relatively well understood. NaCl uptake occurs primarily through the Na ϩ -K ϩ -2Cl Ϫ cotransporter, which has been demonstrated both on functional and transcriptional levels (3,4). The next step is not yet clear. The possible mediators and modulators of the information transmitted between the MD and its target cells include the ion concentration, ATP, angiotensin II, adenosine, arachidonic acid metabolites, and nitric oxide (NO) (5-7). Among these, NO appears to play an important role in the vascular response to changes in luminal [NaCl]. Many studies have been done on the effects of NO on TGF regulation (8 -14), but there has been no report on instantaneous NO concentration changes caused by changes in the luminal [NaCl]. In the present study, an NO-sensitive probe was used for di...

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Section: Discussionmentioning

confidence: 99%

Changes of Cell Volume and Nitric Oxide Concentration in Macula Densa Cells Caused by Changes in Luminal NaCl Concentration

Liu

Pittner

Persson

2002

Journal of the American Society of Nephrology

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“…The same results were found at 2 hr after reoxygenation (data not shown), suggesting that the decrease in MEQ fluorescence 1 and 2 hr after OGD was attributable to an accumulation in intracellular Cl Ϫ . We also measured changes in calcein fluorescence, which is sensitive to changes in cell volume (Crowe et al, 1995;Inglefield and Schwartz-Bloom, 1998a). The calcein ⌬F/F at 2 hr after OGD did not differ from control slices (Fig.…”

Section: Effect Of Ogd On Meq Fluorescencementioning

confidence: 95%

“…To determine changes in cell volume after OGD, slices were bathloaded with calcein acetoxymethyl ester (AM; Molecular Probes) (Crowe et al, 1995;Inglefield and Schwartz-Bloom, 1998a). This indicator is insensitive to changes in intracellular ions.…”

Section: Schwartzmentioning

confidence: 99%

Changes in Intracellular Chloride after Oxygen–Glucose Deprivation of the Adult Hippocampal Slice: Effect of Diazepam

Galeffi¹,

Sah²,

Pond³

et al. 2004

J. Neurosci.

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“…To convert fluorescence values to relative cell volume, the following equation was applied (Capo-Aponte et al, 2005;Crowe et al, 1995;Hamann et al, 2002) where V t is the cell volume at time t, V 0 is initial volume at time = 0, F 0 is the fluorescence of cells in isotonic solution at time = 0 (π 0 ), F t is the fluorescence measured subsequent to a hypotonic challenge at time t (π t ), and f b is the background fluorescence, which represents the fraction of intracellular calcein insensitive to external osmolarity (i.e. f b <1).…”

Section: Cell Volume Measurement and Rvd Analysismentioning

confidence: 99%

Differential dependence of regulatory volume decrease behavior in rabbit corneal epithelial cells on MAPK superfamily activation

Pan¹,

Capó-Aponte²,

Zhang³

et al. 2007

Experimental Eye Research

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We characterized the dependence of hypotonicity-induced regulatory volume decrease (RVD) responses on mitogen-activated protein kinase (MAPK) pathway signaling in SV40-immortalized rabbit corneal epithelial cells (RCEC). Following calcein-AM loading, RVD was monitored using a microplate fluorescence reader. Western blot analysis determined MAPK activation. After 30 min, the RVD response restored the relative cell volume to nearly isotonic values, whereas it was inhibited when cells were bathed either in a Cl − -free solution or with the Cl − -channel inhibitors: 5-nitro-2-(3-phenylpropylamino) benzoic acid or niflumic acid. Similar declines occurred with either a high-K + (20 mM) supplemented solution or the K + channel inhibitor 4-aminopyridine. Activation of extracellular signal-regulated kinase (ERK), p38, and stress-activated protein kinase/c-Jun Nterminal kinase (SAPK/JNK) was time and tonicity-dependent. Stimulation of ERK and SAPK/JNK was maximized earlier than that of p38. Activation of ERK and SAPK/JNK was insensitive to Cl − and K + channel inhibitors, whereas inhibition with either PD98059 or SP600125, respectively, blocked RVD. However, inhibition of p38 with SB203580 had no effect on RVD. Suppression of RVD instead blocked p38 activation. Differences in the dependence of RVD activation on Erk1/2 and p38 signaling were validated in dominant negative (d/n) -Erk1 and d/n-p38 cells. Volumesensitive Cl − and K + channel activation contributes, in concert, to RVD in RCEC. Therefore, swelling-induced ERK and SAPK/JNK stimulation precedes Cl − and K + channel activation, whereas p38 activation occurs as a consequence of RVD.

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Volume changes in single N1E-115 neuroblastoma cells measured with a fluorescent probe

Cited by 111 publications

References 31 publications

Changes of Cell Volume and Nitric Oxide Concentration in Macula Densa Cells Caused by Changes in Luminal NaCl Concentration

Changes of Cell Volume and Nitric Oxide Concentration in Macula Densa Cells Caused by Changes in Luminal NaCl Concentration

Changes in Intracellular Chloride after Oxygen–Glucose Deprivation of the Adult Hippocampal Slice: Effect of Diazepam

Differential dependence of regulatory volume decrease behavior in rabbit corneal epithelial cells on MAPK superfamily activation

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