메뉴 건너뛰기




Volumn 18, Issue 17, 1998, Pages 6882-6891

Thrombin perturbs neurite outgrowth and induces apoptotic cell death in enriched chick spinal motoneuron cultures through caspase activation

Author keywords

Apoptosis; Caspases; PAR 1; Serine proteases; Spinal motoneuron cultures; Thrombin

Indexed keywords

SERINE PROTEINASE; THROMBIN;

EID: 17344365478     PISSN: 02706474     EISSN: None     Source Type: Journal    
DOI: 10.1523/jneurosci.18-17-06882.1998     Document Type: Article
Times cited : (114)

References (59)
  • 1
    • 0023838532 scopus 로고
    • Immunochemical identification of the serine protease inhibitor alpha-1-antichymotrypsin in the brain amyloid deposits of Alzheimer's disease
    • Abraham CR, Selkoe D, Potter H (1988) Immunochemical identification of the serine protease inhibitor alpha-1-antichymotrypsin in the brain amyloid deposits of Alzheimer's disease. Cell 52:487-501.
    • (1988) Cell , vol.52 , pp. 487-501
    • Abraham, C.R.1    Selkoe, D.2    Potter, H.3
  • 2
    • 0025514410 scopus 로고
    • Survival effect of ciliary neurotrophic factor (CNTF) on chick motoneurons in culture: Comparison with other neurotrophic factors and cytokines
    • Arakawa Y, Scndtner M, Thoenen H (1990) Survival effect of ciliary neurotrophic factor (CNTF) on chick motoneurons in culture: comparison with other neurotrophic factors and cytokines. J Neurosci 10:3507-3515.
    • (1990) J Neurosci , vol.10 , pp. 3507-3515
    • Arakawa, Y.1    Scndtner, M.2    Thoenen, H.3
  • 3
    • 0018932068 scopus 로고
    • Proteasenexin: A cellular component that links thrombin and plasminogen activator and mediates their binding to cells
    • Baker JB, Low DA, Simmer RL, Cunningham DD (1980) Proteasenexin: a cellular component that links thrombin and plasminogen activator and mediates their binding to cells. Cell 21:37-45.
    • (1980) Cell , vol.21 , pp. 37-45
    • Baker, J.B.1    Low, D.A.2    Simmer, R.L.3    Cunningham, D.D.4
  • 4
    • 0029760923 scopus 로고    scopus 로고
    • Forging a path to cell death
    • Barinaga M (1996) Forging a path to cell death. Science 273:735-737.
    • (1996) Science , vol.273 , pp. 735-737
    • Barinaga, M.1
  • 5
    • 0028085095 scopus 로고
    • Thrombin receptor peptides induce shape changes in neonatal murine astrocytes in culture
    • Beecher KL, Anderson TT, Fenton II JW, Festoff BW (1994) Thrombin receptor peptides induce shape changes in neonatal murine astrocytes in culture. J Neurosci Res 37:108-115.
    • (1994) J Neurosci Res , vol.37 , pp. 108-115
    • Beecher, K.L.1    Anderson, T.T.2    Fenton II, J.W.3    Festoff, B.W.4
  • 6
    • 0019440757 scopus 로고
    • Regional pattern of degeneration in Alzheimer's disease: Neuronal loss and histopathological grading
    • Brun A, Englund E (1981) Regional pattern of degeneration in Alzheimer's disease: neuronal loss and histopathological grading. Histopathology 5:549-564.
    • (1981) Histopathology , vol.5 , pp. 549-564
    • Brun, A.1    Englund, E.2
  • 7
    • 0025255579 scopus 로고
    • Reciprocal modulation of astrocyte stellation by thrombin and protease nexin-1
    • Cavanaugh K, Gurwitz D, Cunningham DD, Bradshaw R (1990) Reciprocal modulation of astrocyte stellation by thrombin and protease nexin-1. J Neurochem 54:1735-1743.
    • (1990) J Neurochem , vol.54 , pp. 1735-1743
    • Cavanaugh, K.1    Gurwitz, D.2    Cunningham, D.D.3    Bradshaw, R.4
  • 8
    • 0017928027 scopus 로고
    • Cell death of motoneurons in the chick embryo spinal cord. II. A quantitative and qualitative analysis of degeneration in the ventral root, including evidence for axon outgrowth and limb innervation prior to cell death
    • Chu-Wang IW, Oppenheim RW (1978) Cell death of motoneurons in the chick embryo spinal cord. II. A quantitative and qualitative analysis of degeneration in the ventral root, including evidence for axon outgrowth and limb innervation prior to cell death. J Comp Neurol 177:59-85.
    • (1978) J Comp Neurol , vol.177 , pp. 59-85
    • Chu-Wang, I.W.1    Oppenheim, R.W.2
  • 9
    • 0023472509 scopus 로고
    • Neuron numbers and dendritic extent in normal aging and Alzheimer's disease
    • Coleman PD, Flood DG (1987) Neuron numbers and dendritic extent in normal aging and Alzheimer's disease. Neurobiol Aging 8:521-545.
    • (1987) Neurobiol Aging , vol.8 , pp. 521-545
    • Coleman, P.D.1    Flood, D.G.2
  • 12
    • 0023003776 scopus 로고
    • Muscle derived factors that support and promote fiber outgrowth from embryonic chick spinal motor neurons in culture
    • Dohrman U, Edgard D, Sendtner M, Thoenen H (1986) Muscle derived factors that support and promote fiber outgrowth from embryonic chick spinal motor neurons in culture. Dev Biol 118:209-211.
    • (1986) Dev Biol , vol.118 , pp. 209-211
    • Dohrman, U.1    Edgard, D.2    Sendtner, M.3    Thoenen, H.4
  • 13
    • 16944366731 scopus 로고    scopus 로고
    • Thrombin induces apoptosis in cultured neurons and astrocytes via a pathway requiring tyrosine kinase and RhoA activities
    • Donovan FM, Pike CJ, Cunningham DD (1997) Thrombin induces apoptosis in cultured neurons and astrocytes via a pathway requiring tyrosine kinase and RhoA activities. J Neurosci 17:5316-5326.
    • (1997) J Neurosci , vol.17 , pp. 5316-5326
    • Donovan, F.M.1    Pike, C.J.2    Cunningham, D.D.3
  • 14
    • 0026638915 scopus 로고
    • Early stages of motor neuron differentiation revealed by expression of homeobox gene Islet-1
    • Ericson J, Thor S, Edlund T, Jessell TM, Yamada T (1992) Early stages of motor neuron differentiation revealed by expression of homeobox gene Islet-1. Science 256:1555-1560.
    • (1992) Science , vol.256 , pp. 1555-1560
    • Ericson, J.1    Thor, S.2    Edlund, T.3    Jessell, T.M.4    Yamada, T.5
  • 16
    • 0030007646 scopus 로고    scopus 로고
    • Thrombin, its receptor and protease nexin I, its potent serpin, in the nervous system
    • Festoff BW, Smirnova IV, Ma J, Citron BA (1996) Thrombin, its receptor and protease nexin I, its potent serpin, in the nervous system. Semin Thromb Hemost 22:267-271.
    • (1996) Semin Thromb Hemost , vol.22 , pp. 267-271
    • Festoff, B.W.1    Smirnova, I.V.2    Ma, J.3    Citron, B.A.4
  • 17
    • 0026648674 scopus 로고
    • Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation
    • Gavrieli Y, Sherman Y, Ben-Saan SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119:493-501.
    • (1992) J Cell Biol , vol.119 , pp. 493-501
    • Gavrieli, Y.1    Sherman, Y.2    Ben-Saan, S.A.3
  • 18
    • 0026729942 scopus 로고
    • Vitronectin is the major serum protein essential for NGF-mediated neurite outgrowth from PC12 cells
    • Grabham PW, Gallimore PH, Grand RJ (1992) Vitronectin is the major serum protein essential for NGF-mediated neurite outgrowth from PC12 cells. Exp Cell Res 202:337-344.
    • (1992) Exp Cell Res , vol.202 , pp. 337-344
    • Grabham, P.W.1    Gallimore, P.H.2    Grand, R.J.3
  • 19
    • 0011811083 scopus 로고
    • Thrombin modulates and reverses neuroblastoma neurite outgrowth
    • Gurwitz D, Cunningham DD (1988) Thrombin modulates and reverses neuroblastoma neurite outgrowth. Proc Natl Acad Sci USA 85:3440-3444.
    • (1988) Proc Natl Acad Sci USA , vol.85 , pp. 3440-3444
    • Gurwitz, D.1    Cunningham, D.D.2
  • 20
    • 0025189336 scopus 로고
    • Neurite outgrowth activity of protease nexin-1 on neuroblastoma cells requires thrombin inhibition
    • Gurwitz D, Cunningham DD (1990) Neurite outgrowth activity of protease nexin-1 on neuroblastoma cells requires thrombin inhibition. J Cell Physiol 142:155-162.
    • (1990) J Cell Physiol , vol.142 , pp. 155-162
    • Gurwitz, D.1    Cunningham, D.D.2
  • 21
    • 84924926133 scopus 로고
    • A series of normal stages in the development of the chick embryo
    • Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49-92.
    • (1951) J Morphol , vol.88 , pp. 49-92
    • Hamburger, V.1    Hamilton, H.L.2
  • 23
    • 0028800603 scopus 로고
    • A serine protease inhibitor, protease nexin 1, rescues motoneurons from naturally occurring and axotomy-induced cell death
    • Houenou LJ, Turner PL, Li L, Oppenheim RW, Festoff BW (1995) A serine protease inhibitor, protease nexin 1, rescues motoneurons from naturally occurring and axotomy-induced cell death. Proc Natl Acad Sci USA 92:895-899.
    • (1995) Proc Natl Acad Sci USA , vol.92 , pp. 895-899
    • Houenou, L.J.1    Turner, P.L.2    Li, L.3    Oppenheim, R.W.4    Festoff, B.W.5
  • 24
    • 0027309290 scopus 로고
    • Kinetics of thrombin receptor cleavage on intact cells. Relation to signaling
    • Ishii K, Hein L, Kobilka B, Coughlin SR (1993) Kinetics of thrombin receptor cleavage on intact cells. Relation to signaling. J Biol Chem 268:9780-9786.
    • (1993) J Biol Chem , vol.268 , pp. 9780-9786
    • Ishii, K.1    Hein, L.2    Kobilka, B.3    Coughlin, S.R.4
  • 25
    • 0026058318 scopus 로고
    • Cell death during development of the nervous system
    • Oppenheim RW (1991) Cell death during development of the nervous system. Annu Rev Neurosci 14:453-501.
    • (1991) Annu Rev Neurosci , vol.14 , pp. 453-501
    • Oppenheim, R.W.1
  • 26
    • 0023517177 scopus 로고
    • Thrombin is a potent mitogen for rat astroblasts but not for oligodendroblasts and neuroblasts in primary culture
    • Perraud F, Besnard F, Sensenbrenner M, Labourdette G (1987) Thrombin is a potent mitogen for rat astroblasts but not for oligodendroblasts and neuroblasts in primary culture. Int J Dev Neurosci 5:181-188.
    • (1987) Int J Dev Neurosci , vol.5 , pp. 181-188
    • Perraud, F.1    Besnard, F.2    Sensenbrenner, M.3    Labourdette, G.4
  • 27
    • 0027957279 scopus 로고
    • Increased levels of plasminogen activator inhibitor-1 (PA-1) in human brain tumors
    • Rao JS, Rayford A, Mortantz RA, Festoff BW, Sawaya R (1993) Increased levels of plasminogen activator inhibitor-1 (PA-1) in human brain tumors. J Neuro-Oncol 17:215:221.
    • (1993) J Neuro-oncol , vol.17 , pp. 215
    • Rao, J.S.1    Rayford, A.2    Mortantz, R.A.3    Festoff, B.W.4    Sawaya, R.5
  • 28
    • 0027976526 scopus 로고
    • Glia-derived nexin/ protease nexin-1 is expressed by a subset of neurons in the rat brain
    • Reinhard E, Suidan HS, Pavlik A, Monard D (1994) Glia-derived nexin/ protease nexin-1 is expressed by a subset of neurons in the rat brain. J Neurosci Res 37:256-270.
    • (1994) J Neurosci Res , vol.37 , pp. 256-270
    • Reinhard, E.1    Suidan, H.S.2    Pavlik, A.3    Monard, D.4
  • 30
    • 0031823351 scopus 로고    scopus 로고
    • Quantitative reversetranscription PCR to gauge increased protease-activated receptor 1 (PAR-1) mRNA copy numbers in the wobbler mutant mouse
    • in press
    • Salcedo RM, Festoff BW, Citron BA (1998) Quantitative reversetranscription PCR to gauge increased protease-activated receptor 1 (PAR-1) mRNA copy numbers in the wobbler mutant mouse. J Mol Neurosci, in press.
    • (1998) J Mol Neurosci
    • Salcedo, R.M.1    Festoff, B.W.2    Citron, B.A.3
  • 31
    • 0026520206 scopus 로고
    • Involvement of the "tethered-ligand" receptor in thrombin inhibition of platelet adenylate cyclase
    • Seiler SM, Michel IM, Fenton II JW (1992) Involvement of the "tethered-ligand" receptor in thrombin inhibition of platelet adenylate cyclase. Biochem Res Commun 182:1296-1302.
    • (1992) Biochem Res Commun , vol.182 , pp. 1296-1302
    • Seiler, S.M.1    Michel, I.M.2    Fenton II, J.W.3
  • 32
    • 0029002271 scopus 로고
    • Inhibition of neuronal surface proteases decreases the requirement for GAP-43 in neurite outgrowth
    • Shea TB (1995) Inhibition of neuronal surface proteases decreases the requirement for GAP-43 in neurite outgrowth. Dev Brain Res 87:87-90.
    • (1995) Dev Brain Res , vol.87 , pp. 87-90
    • Shea, T.B.1
  • 33
    • 0029159809 scopus 로고
    • Enhancement of neurite outgrowth following calpain inhibition is mediated by protein kinase C
    • Shea TB, Cressman CM, Spencer MJ, Beerman ML, Nixon RA (1995) Enhancement of neurite outgrowth following calpain inhibition is mediated by protein kinase C. J Neurochem 65:517-527.
    • (1995) J Neurochem , vol.65 , pp. 517-527
    • Shea, T.B.1    Cressman, C.M.2    Spencer, M.J.3    Beerman, M.L.4    Nixon, R.A.5
  • 34
    • 0024363137 scopus 로고
    • Calpain I activation is specifically related to excitatory amino acid induction of hippocampal damage
    • Siman R, Noszek JC, Kegerise C (1989) Calpain I activation is specifically related to excitatory amino acid induction of hippocampal damage. J Neurosci 9:1579-1590.
    • (1989) J Neurosci , vol.9 , pp. 1579-1590
    • Siman, R.1    Noszek, J.C.2    Kegerise, C.3
  • 35
    • 0028027469 scopus 로고
    • Extravascular proteolysis and the nervous system: Serine protease/serpin balance
    • Smirnova IV, Ho GJ, Fenton II JW, Festoff BW (1994) Extravascular proteolysis and the nervous system: serine protease/serpin balance. Semin Thromb Hemost 20:426-432.
    • (1994) Semin Thromb Hemost , vol.20 , pp. 426-432
    • Smirnova, I.V.1    Ho, G.J.2    Fenton II, J.W.3    Festoff, B.W.4
  • 36
    • 17344370251 scopus 로고    scopus 로고
    • Thrombin is a death signal that activates intracellular death protease pathways in motor neurons
    • in press
    • Smirnova IV, Zhang SX, Citron BA, Arnold PM, Festoff BW (1998a) Thrombin is a death signal that activates intracellular death protease pathways in motor neurons. J Neurobiol, in press.
    • (1998) J Neurobiol
    • Smirnova, I.V.1    Zhang, S.X.2    Citron, B.A.3    Arnold, P.M.4    Festoff, B.W.5
  • 37
    • 0031814260 scopus 로고    scopus 로고
    • Calcium mobilization and protease-activated receptor cleavage after thrombin stimulation in motor neurons
    • in press
    • Smirnova IV, Vamos S, Wiegmann T, Citron BA, Arnold PM, Festoff BW (1998b) Calcium mobilization and protease-activated receptor cleavage after thrombin stimulation in motor neurons. J Mol Neurosci, in press.
    • (1998) J Mol Neurosci
    • Smirnova, I.V.1    Vamos, S.2    Wiegmann, T.3    Citron, B.A.4    Arnold, P.M.5    Festoff, B.W.6
  • 40
    • 0029830345 scopus 로고    scopus 로고
    • Induction of CPP32 like activity in PC12 cells by withdrawal of trophic support: Dissociation from apoptosis
    • Stefanis L, Park DS, Yan CYI, Farinelli SE, Troy CM, Shelanski ML, Greene LA (1996) Induction of CPP32 like activity in PC12 cells by withdrawal of trophic support: dissociation from apoptosis. J Biol Chem 271:30663-30671.
    • (1996) J Biol Chem , vol.271 , pp. 30663-30671
    • Stefanis, L.1    Park, D.S.2    Yan, C.Y.I.3    Farinelli, S.E.4    Troy, C.M.5    Shelanski, M.L.6    Greene, L.A.7
  • 41
    • 0030984171 scopus 로고    scopus 로고
    • Inhibitors of trypsin-like serine proteases inhibit processing of the caspase nedd-2 and protect PC12 cells and sympathetic neurons from death evoked by withdrawal of trophic support
    • Stefanis L, Troy CM, Haiqing Q, Greene LA (1997) Inhibitors of trypsin-like serine proteases inhibit processing of the caspase nedd-2 and protect PC12 cells and sympathetic neurons from death evoked by withdrawal of trophic support. J Neurochem 69:1425-1437.
    • (1997) J Neurochem , vol.69 , pp. 1425-1437
    • Stefanis, L.1    Troy, C.M.2    Haiqing, Q.3    Greene, L.A.4
  • 42
    • 0022521744 scopus 로고
    • Kinetics of the inhibition of thrombin by hirudin
    • Stone SR, Hofsteenge J (1986) Kinetics of the inhibition of thrombin by hirudin. Biochemistry 25:4622-4628.
    • (1986) Biochemistry , vol.25 , pp. 4622-4628
    • Stone, S.R.1    Hofsteenge, J.2
  • 43
    • 0026706235 scopus 로고
    • The inhibition of thrombin and chymotrypsin by heparin-cofactor III
    • Struss D, Storck J, Zimmerman RE (1992) The inhibition of thrombin and chymotrypsin by heparin-cofactor III. Thomb Res 68:45-56.
    • (1992) Thomb Res , vol.68 , pp. 45-56
    • Struss, D.1    Storck, J.2    Zimmerman, R.E.3
  • 44
    • 0026519282 scopus 로고
    • Thrombin causes neurite retraction in neuronal cells through activation of cell surface receptors
    • Suidan HS, Stone SR, Hemmings BA, Monard D (1992) Thrombin causes neurite retraction in neuronal cells through activation of cell surface receptors. Neuron 8:363-375.
    • (1992) Neuron , vol.8 , pp. 363-375
    • Suidan, H.S.1    Stone, S.R.2    Hemmings, B.A.3    Monard, D.4
  • 45
    • 0026472983 scopus 로고
    • What do we really know about amyotrophic lateral sclerosis?
    • Swash M, Schwartz MS (1992) What do we really know about amyotrophic lateral sclerosis? J Neural Sci 113:4-16.
    • (1992) J Neural Sci , vol.113 , pp. 4-16
    • Swash, M.1    Schwartz, M.S.2
  • 46
    • 0024542079 scopus 로고
    • A new membrane antigen revealed by monoclonal antibodies is associated with motoneuron axonal pathways
    • Tanaka H, Agata A, Obata K (1989) A new membrane antigen revealed by monoclonal antibodies is associated with motoneuron axonal pathways. Dev Biol 132:419-435.
    • (1989) Dev Biol , vol.132 , pp. 419-435
    • Tanaka, H.1    Agata, A.2    Obata, K.3
  • 48
    • 0029088484 scopus 로고
    • Excitotoxin-induced neuronal degeneration and seizure are mediated by tissue plasminogen activator
    • Tsirka SE, Gualandris A, Amaral DG, Strickland S (1995) Excitotoxin-induced neuronal degeneration and seizure are mediated by tissue plasminogen activator. Nature 377:340-344.
    • (1995) Nature , vol.377 , pp. 340-344
    • Tsirka, S.E.1    Gualandris, A.2    Amaral, D.G.3    Strickland, S.4
  • 49
    • 0030829885 scopus 로고    scopus 로고
    • The role of thrombin-like (serine) proteases in the development, plasticity and pathology of the nervous system
    • Turgeon VL, Houenou LJ (1997) The role of thrombin-like (serine) proteases in the development, plasticity and pathology of the nervous system. Brain Res Rev 25:85-95.
    • (1997) Brain Res Rev , vol.25 , pp. 85-95
    • Turgeon, V.L.1    Houenou, L.J.2
  • 50
    • 0027468772 scopus 로고
    • Regulation of protease nexin-1 synthesis and secretion in cultured brain cells by injury-related factors
    • Vaughan PJ, Cunningham DD (1993) Regulation of protease nexin-1 synthesis and secretion in cultured brain cells by injury-related factors. J Biol Chem 268:3720-3727.
    • (1993) J Biol Chem , vol.268 , pp. 3720-3727
    • Vaughan, P.J.1    Cunningham, D.D.2
  • 51
    • 0029012029 scopus 로고
    • Thrombin receptor activation protects neurons and astrocytes from cell death produced by environmental insults
    • Vaughan PJ, Pike CJ, Cotman CW, Cunningham DD (1995) Thrombin receptor activation protects neurons and astrocytes from cell death produced by environmental insults. J Neurosci 15:5389-5401.
    • (1995) J Neurosci , vol.15 , pp. 5389-5401
    • Vaughan, P.J.1    Pike, C.J.2    Cotman, C.W.3    Cunningham, D.D.4
  • 52
    • 0026035523 scopus 로고
    • Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation
    • Vu TH, Hung DT, Wheaton VI, Coughlin SR (1991) Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 64:1057-1068.
    • (1991) Cell , vol.64 , pp. 1057-1068
    • Vu, T.H.1    Hung, D.T.2    Wheaton, V.I.3    Coughlin, S.R.4
  • 55
    • 0028914068 scopus 로고
    • Cellular localization of thrombin receptor mRNA in rat brain: Expression by mesencephalic dopaminergic neurons and codistribution with prothrombin mRNA
    • Weinstein JR, Gold SJ, Cunningham DD, Gall CM (1995) Cellular localization of thrombin receptor mRNA in rat brain: expression by mesencephalic dopaminergic neurons and codistribution with prothrombin mRNA. J Neurosci 15:2906-2919.
    • (1995) J Neurosci , vol.15 , pp. 2906-2919
    • Weinstein, J.R.1    Gold, S.J.2    Cunningham, D.D.3    Gall, C.M.4
  • 57
    • 0001918396 scopus 로고
    • Cell death: A new classification separating apoptosis from necrosis
    • (Bowen ID, Lockstein RA, eds) . New York: Chapman and Hall
    • Wyllie AH (1981) Cell death: a new classification separating apoptosis from necrosis. In: Cell death in biology and pathology (Bowen ID, Lockstein RA, eds), pp 9-34. New York: Chapman and Hall.
    • (1981) Cell Death in Biology and Pathology , pp. 9-34
    • Wyllie, A.H.1
  • 58
    • 0025071270 scopus 로고
    • Differential vulnerability of microtubule components in cerebral ischemia
    • Yanagihara Y, Brengman JM, Mushynski WE (1990) Differential vulnerability of microtubule components in cerebral ischemia. Acta Neuropathol (Berl) 80:499-505.
    • (1990) Acta Neuropathol (Berl) , vol.80 , pp. 499-505
    • Yanagihara, Y.1    Brengman, J.M.2    Mushynski, W.E.3
  • 59
    • 0023851616 scopus 로고
    • A glia-derived nexin promotes neurite outgrowth in cultured chick sympathetic neurons
    • Zurn AD, Nick H, Monard D (1988) A glia-derived nexin promotes neurite outgrowth in cultured chick sympathetic neurons. Dev Neurosci 10:17-24.
    • (1988) Dev Neurosci , vol.10 , pp. 17-24
    • Zurn, A.D.1    Nick, H.2    Monard, D.3


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.