Thrombin receptor on rat primary hippocampal neurons: coupled calcium and cAMP responses

Yang, Yaxiong; Akiyama, Haruhiko; Fenton, John W.; Brewer, Gregory J.

doi:10.1016/s0006-8993(97)00311-9

Cited by 20 publications

(7 citation statements)

References 52 publications

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“…These data suggest that a modest concentration of thrombin (30 n m ) was relatively selective for PAR1 activation, since removal or block of PAR1 eliminated the intracellular Ca 2+ response in cultured astrocytes. These results are consistent with a number of previous reports in the literature suggesting that thrombin stimulates Ca 2+ mobilization in neurones and glia (Smith‐Swintosky et al 1995; Yang et al 1997; Smirnova et al 1998; Wang et al 2002 a , b ; Suo et al 2002; Sorensen et al 2003; Junge et al 2004; Nicole et al 2005). The TFLLR‐induced astrocytic Ca 2+ responses persisted in the absence of external Ca 2+ (Fig.…”

Section: Resultssupporting

confidence: 93%

“…2) as well as with other reports in the literature (e.g. Yang et al 1997), but is lower than our previous estimate of ∼40% of thrombin responsive neurones in cultures prepared from the entire hippocampal formation plus dentate gyrus (13% respond to thrombin + inhibitor, 54% respond to 30 n m thrombin; Gingrich et al 2000). This discrepancy likely reflects higher PAR1 expression in dentate granule neurones than in CA1 neurones (Weinstein et al 1995).…”

Section: Resultssupporting

confidence: 44%

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Astrocytic control of synaptic NMDA receptors

Lee

Mannaioni

Yuan

et al. 2007

The Journal of Physiology

200

243

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Astrocytes express a wide range of G-protein coupled receptors that trigger release of intracellular Ca 2+ , including P2Y, bradykinin and protease activated receptors (PARs). By using the highly sensitive sniffer-patch technique, we demonstrate that the activation of P2Y receptors, bradykinin receptors and protease activated receptors all stimulate glutamate release from cultured or acutely dissociated astrocytes. Of these receptors, we have utilized PAR1 as a model system because of favourable pharmacological and molecular tools, its prominent expression in astrocytes and its high relevance to neuropathological processes. Astrocytic PAR1-mediated glutamate release in vitro is Ca 2+ dependent and activates NMDA receptors on adjacent neurones in culture. Activation of astrocytic PAR1 in hippocampal slices induces an APV-sensitive inward current in CA1 neurones and causes APV-sensitive neuronal depolarization in CA1 neurones, consistent with release of glutamate from astrocytes. PAR1 activation enhances the NMDA receptor-mediated component of synaptic miniature EPSCs, evoked EPSCs and evoked EPSPs in a Mg 2+ -dependent manner, which may reflect spine head depolarization and consequent reduction of NMDA receptor Mg 2+ block during subsequent synaptic currents. The release of glutamate from astrocytes following PAR1 activation may also lead to glutamate occupancy of some perisynaptic NMDA receptors, which pass current following relief of tonic Mg 2+ block during synaptic depolarization. These results suggest that astrocytic G-protein coupled receptors that increase intracellular Ca 2+ can tune synaptic NMDA receptor responses.

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Section: Resultssupporting

confidence: 93%

Section: Resultssupporting

confidence: 44%

Astrocytic control of synaptic NMDA receptors

Lee

Mannaioni

Yuan

et al. 2007

The Journal of Physiology

200

243

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“…Thrombin acted specifically, because it affected neither the distribution of surface L1 nor that of NrCAM bearing a noncleavable GFP-tag ( Figure 1E). Although hippocampal neurons express thrombin receptors (Yang et al, 1997), thrombin can induce significant effects only when applied at much higher concentrations and for longer times than the times used here (Brewer, 1996;Donovan et al, 1997). Indeed, thrombin did not alter growth cone motility or cause neurite retraction in our conditions ( Figure 5).…”

Section: Surface Expression and Exocytosis Of L1-gfp Moleculesmentioning

confidence: 80%

Fast Turnover of L1 Adhesions in Neuronal Growth Cones Involving Both Surface Diffusion and Exo/Endocytosis of L1 Molecules

Dequidt

Danglot

Alberts

et al. 2007

MBoC

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We investigated the interplay between surface trafficking and binding dynamics of the immunoglobulin cell adhesion molecule L1 at neuronal growth cones. Primary neurons were transfected with L1 constructs bearing thrombin-cleavable green fluorescent protein (GFP), allowing visualization of newly exocytosed L1 or labeling of membrane L1 molecules by Quantum dots. Intracellular L1-GFP vesicles showed preferential centrifugal motion, whereas surface L1-GFP diffused randomly, revealing two pathways to address L1 to adhesive sites. We triggered L1 adhesions using microspheres coated with L1-Fc protein or anti-L1 antibodies, manipulated by optical tweezers. Microspheres coupled to the actin retrograde flow at the growth cone periphery while recruiting L1-GFP molecules, of which 50% relied on exocytosis. Fluorescence recovery after photobleaching experiments revealed a rapid recycling of L1-GFP molecules at L1-Fc (but not anti-L1) bead contacts, attributed to a high lability of L1-L1 bonds at equilibrium. L1-GFP molecules truncated in the intracellular tail as well as neuronal cell adhesion molecules (NrCAMs) missing the clathrin adaptor binding sequence showed both little internalization and reduced turnover rates, indicating a role of endocytosis in the recycling of mature L1 contacts at the base of the growth cone. Thus, unlike for other molecules such as NrCAM or N-cadherin, diffusion/trapping and exo/endocytosis events cooperate to allow the fast renewal of L1 adhesions.

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“…Additionally, PAR-I's existence at the neural cell surface has been inferred for some time based on morphologic, physiologic, and biochemical responses to both thrombin and PAR-I activating peptides (Jatink and Moolenaar, 1992;Suidan et al, 1992;Beecher et al, 1994;Grabham and Cunningham, 1995;Smith-Swintosky et al, 1995;Vaughan et at., 1995;Yang et al, 1997). In the present study, PAR-I cell surface expression is directly demonstrated and quantitated in a human neural cell line (Ad12 HER 10).…”

Section: Par-i Cell Surface Expression May Allow For Both Normal Devementioning

confidence: 83%

Injury‐Related Factors and Conditions Down‐Regulate the Thrombin Receptor (PAR‐1) in a Human Neuronal Cell Line

Weinstein

Lau

Brass

et al. 1998

Journal of Neurochemistry

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Jou,uif ot NCUrO('/U'iniStJ'rAbstract: Previous studies have demonstrated that thrombin can induce potent effects on neural cell morphology, biochemistry, and viability. Nearly all of these effects are mediated by proteolytic activation of the thrombin receptor (PAR-i). Mechanisms of PAR-i regulation in several nonneural cell types have been shown to be novel and cell type specific; however, little is known about PAR-i regulation in neural cells. In the present study, PAR-i cell surface expression and regulation were examined in a transformed retinoblast (Adi 2 HER i 0) cell line using radioiodinated anti-PAR-i monoclonal antibodies ATAP2, which recognizes intact and cleaved receptors, and SPANi2, which is specific for the intact form of the receptor. Scatchard analysis revealed high-affinity, specific binding to a single affinity class of receptors: K= 3.13 and 5.25 nM, Bmax = i90.i and 67.8 fmol/mg of protein for 1251-ATAP2 and 125l-SPANi2, respectively. Specificity for PAR-i was confirmed by demonstrating rapid and near complete decreases for both antibodies following treatment with thrombin or PAR-i activating peptide (SFLLRN). Differential antibody binding was used to demonstrate rapid and near complete thrombininduced PAR-i cleavage and internalization, with protein synthesis-dependent replacement of intact receptors occurring over longer time intervals, but only minimal recycling of cleaved receptors. A variety of factors and conditions were screened for their effects on PAR-i expression. Significant decreases in PAR-i expression were induced by the protein kinase C activator phorbol 12-myristate i3-acetate (87% at 3 h), the phospholipid inflammatory mediator lysophosphatidic acid (32% at 3 h), and the injury-related condition hypoglycemia (64 and 100% at 24 h in the absence and presence of dibutyryl cyclic AMP, respectively). The effect of hypoglycemia was shown by RNase protection to be at least partially pretranslational. Finally, thrombin's ability to enhance hypoglycemia-induced cell killing correlated temporally with PAR-i cell surface expression. Key Words: Thrombin receptor-PAR-i -Regulation-Neurons-Lysophosphatidic acid-Hypoglycemia-Antibody. J. Neurochem. 71, i034-i050 (i998).Over the last decade, evidence has accumulated implicating the multifunctional serine protease thrombin in a variety of developmental, physiologic, and pathophysiologic functions in the nervous system (for review, see Grand et al., 1996). Thrombin can induce a variety of effects on cultured neural cells, including neurite retraction (Gurwitz and Cunningham, 1988; Grand et al., 1989;Farmer et al., 1990), reversal of astrocyte stellation (Cavanaugh et al

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Thrombin receptor on rat primary hippocampal neurons: coupled calcium and cAMP responses

Cited by 20 publications

References 52 publications

Astrocytic control of synaptic NMDA receptors

Astrocytic control of synaptic NMDA receptors

Fast Turnover of L1 Adhesions in Neuronal Growth Cones Involving Both Surface Diffusion and Exo/Endocytosis of L1 Molecules

Injury‐Related Factors and Conditions Down‐Regulate the Thrombin Receptor (PAR‐1) in a Human Neuronal Cell Line

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