Essential role of myosin light chain kinase in the mechanism for MgATP- dependent priming of exocytosis in adrenal chromaffin cells

Kumakura, Konosuke; Sasaki, Kimihito; Sakurai, Takashi; Ohara‐Imaizumi, Mica; Misonou, Hiroaki; Nakamura, Seiji; Matsuda, Yuzuru; Nonomura, Yoshiaki

doi:10.1523/jneurosci.14-12-07695.1994

Cited by 85 publications

(46 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Myosin light chain kinase, which is a key regulator of myosin II, has also been implicated in vesicle release at neural synapses ( 22 ) and the ATP-dependent priming of exocytosis in chromaffi n cells ( 23 ), further indicating that myosin II may play a key role in vesicle fusion.…”

Section: Cells and Inhibitorsmentioning

confidence: 99%

Myosin IIA is required for cytolytic granule exocytosis in human NK cells

Andzelm¹,

Chen²,

Krzewski³

et al. 2007

J Cell Biol

View full text Add to dashboard Cite

Section: Cells and Inhibitorsmentioning

confidence: 99%

Myosin IIA is required for cytolytic granule exocytosis in human NK cells

Andzelm¹,

Chen²,

Krzewski³

et al. 2007

J Cell Biol

View full text Add to dashboard Cite

“…Acetylcholine-SNAP and NSF were also shown to increase the priming rate (Xu et al, 1999). Several kinases such as PKA, protein kinase C (PKC), Ca 2ϩ -regulated myosin light chain kinase have been shown to modulate neurotransmitter release (Kumakura et al, 1994;Capogna et al, 1995;Mochida, 1995). Finally, PtdIns(4,5)P 2 formation by the conjoint ATP-dependent activities of PI4K and PI4P5K has been shown to be required for LDCV priming (Martin et al, 1997) In contrast to its role in signal transduction, where it acts as a substrate for the generation of diacylglycerol and InsP 3 , PtdIns(4,5)P 2 itself is likely to serve as a membrane anchor or modulator for several proteins of the secretory apparatus, such as synaptotagmins and CAPS (Walent et al, 1992;Schiavo et al, 1996).…”

Section: Involvement Of Pi3k-c2␣ Enzymatic Activity In Neurosecretionmentioning

confidence: 99%

Phosphatidylinositol 3-Kinase C2α Is Essential for ATP-dependent Priming of Neurosecretory Granule Exocytosis

Meunier

Osborne

Hammond

et al. 2005

MBoC

102

108

View full text Add to dashboard Cite

Neurotransmitter release and hormonal secretion are highly regulated processes culminating in the calcium-dependent fusion of secretory vesicles with the plasma membrane. Here, we have identified a role for phosphatidylinositol 3-kinase C2␣ (PI3K-C2␣) and its main catalytic product, PtdIns3P, in regulated exocytosis. In neuroendocrine cells, PI3K-C2␣ is present on a subpopulation of mature secretory granules. Impairment of PI3K-C2␣ function specifically inhibits the ATP-dependent priming phase of exocytosis. Overexpression of wild-type PI3K-C2␣ enhanced secretion, whereas transfection of PC12 cells with a catalytically inactive PI3K-C2␣ mutant or a 2xFYVE domain sequestering PtdIns3P abolished secretion. Based on these results, we propose that production of PtdIns3P by PI3K-C2␣ is required for acquisition of fusion competence in neurosecretion. INTRODUCTIONThe analysis of the molecular mechanism controlling neuroexocytosis has been greatly helped by studies on neurosecretory cells (Dunn and Holz, 1983;Sarafian et al., 1987;Martin and Walent, 1989;Monck and Fernandez, 1994;Rettig and Neher, 2002). Early work has shed light on two major steps consisting of the reversible ATP-dependent priming of docked granules followed by their Ca 2ϩ -driven fusion with the plasma membrane and release of the granule content (Holz et al., 1989;Hay and Martin, 1992). The spatiotemporal control of intracellular Ca 2ϩ concentration together with capacitance measurements has allowed the identification of distinct pools of chromaffin granules involved in these distinct steps (Rettig and Neher, 2002). Further electrophysiological and biochemical approaches, such as the reconstitution of secretion in permeabilized neurosecretory cells (Martin and Walent, 1989), have highlighted the role of key factors involved in these two processes. Cytosolic and membrane proteins have been linked to priming and fusion, including the mammalian orthologues of the Caenorhabditis elegans UNC13 and UNC18 gene products, synaptotagmins, and members of the SNARE family and associated proteins (Jahn and Sudhof, 1999;Burgoyne et al., 2001;Brose and Rosenmund, 2002;Rettig and Neher, 2002).In contrast, less is known about the contribution of lipid dynamics during these processes with the exception of phosphatidic acid and phosphatidylinositol (4,5)-bisphosphate [PtdIns(4,5)P 2 ]. The former is produced by phospholipase D, which is essential for secretion in neurons (Humeau et al., 2001) and in neuroendocrine cells . The inhibition of Ca 2ϩ -dependent catecholamine release after depletion of phosphatidylinositol highlighted the role of phosphoinositides during the secretory events (Eberhard et al., 1990). Moreover, phosphatidylinositol transfer protein and phosphatidylinositol-4-phosphate 5 kinase (PI4P5K) were shown to be required for the ATP-dependent priming of secretory granules in PC12 cells (Hay and Martin, 1993;Hay et al., 1995). In addition, phosphatidylinositol 4-kinase (PI4K), an integral membrane protein of chromaffin granules and synaptic vesicles, is r...

show abstract

“…However, the e ective concentration range of wortmannin for inhibiting the release of SP from cultured DRG neurons was higher than that inhibiting PI3-kinase. Furthermore, wortmannin showed similar IC 50 values (approximately 1 mM) for inhibition of the release of catecholamines in both intact and permealized chroma n cells (Ohara-Imaizumi et al, 1992;Kumakura et al, 1994), suggesting that cell membranes do not hamper the penetration of wortmannin to the interior of neurons. Taken together, the e ect of wortmannin in the present study suggests that phosphorylation of myosin II plays an important role in the release of SP.…”

Section: Discussionmentioning

confidence: 93%

“…The anti-SP antibody was also reported to react speci®cally with SP in the rat spinal cord (Cuello et al, 1979), but not with other peptides such as Leuor Met-enkephalin (see manufacturer's product description). Characterization of the anti-myosin II rabbit antibody was described elsewhere (Kumakura et al, 1994;Mochida et al, 1994). Immunoblotting of homogenates from various kinds of tissues including nervous system showed that the antibody binds to a single band of 200 KDa, which corresponds to the molecular weight of mysoin II heavy chain.…”

Section: Immuno¯uorescent Experimentsmentioning

confidence: 99%

“…Recent studies by Nonomura and colleagues showed that wortmannin, an inhibitor of myosin light chain kinase (MLCK), exerts inhibitory e ects on various exocytic events: histamine secretion from rat basophilic leukaemia (RBL-2H3) cells (Kitani et al, 1992;Ozawa et al, 1996), catecholamine secretion from bovine adrenal chroma n cells (Ohara-Imaizumi et al, 1992;Kumakura et al, 1994), insulin secretion from pancreatic b cell line MIN6 (Hagiwara et al, 1995), and the release of human immunode®ciency virus type 1 from host cells (Sasaki et al, 1995). These results suggest that phosphorylation of myosin II is involved in these exocytic events.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Differential effects of wortmannin on the release of substance P and amino acids from the isolated spinal cord of the neonatal rat

Suzuki

Yoshioka

Maehara

et al. 1998

British J Pharmacology

View full text Add to dashboard Cite

1 E ects of wortmannin, an inhibitor of myosin light chain kinase, on the release of substance P and amino acids, GABA and glutamate, were investigated in the isolated spinal cord preparation of the neonatal rat. 2 Wortmannin at 0.5 ± 10 mM depressed the release of substance P evoked by high-K + (90 mM) medium from the spinal cord (IC 50 =1.1 mM). Wortmannin also depressed the high-K + (70 mM)-evoked release of substance P from cultured dorsal root ganglion neurons of neonatal rats. In contrast, the high-K + (90 mM)-evoked release of GABA and glutamate from the spinal cord was not a ected by wortmannin (0.1 ± 10 mM). 3 Upon stimulation of a dorsal root, a monosynaptic re¯ex and a subsequent slow ventral root depolarization were evoked in the ipsilateral ventral root of the same segment in the isolated spinal cord preparation. The magnitude of the slow ventral root depolarization was depressed gradually to about 70% of the control during the course of 30 min under wortmannin (1 mM). In contrast, the monosynaptic re¯ex was una ected by wortmannin. 4 Immuno¯uorescent staining revealed that immunoreactivities of substance P and myosin II were colocalized at presynaptic terminals in the dorsal horn of the neonatal rat spinal cord. 5 The present results suggest that myosin phosphorylation by myosin light chain kinase may play a crucial role in the release of substance P, but not in the release of GABA and glutamate in the neonatal rat spinal cord. This may re¯ect a di erence in the exocytic mechanisms of substance Pcontaining large dense core vesicles and amino acid-containing small clear vesicles.

show abstract

Essential role of myosin light chain kinase in the mechanism for MgATP- dependent priming of exocytosis in adrenal chromaffin cells

Cited by 85 publications

References 51 publications

Myosin IIA is required for cytolytic granule exocytosis in human NK cells

Myosin IIA is required for cytolytic granule exocytosis in human NK cells

Phosphatidylinositol 3-Kinase C2α Is Essential for ATP-dependent Priming of Neurosecretory Granule Exocytosis

Differential effects of wortmannin on the release of substance P and amino acids from the isolated spinal cord of the neonatal rat

Contact Info

Product

Resources

About