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Detection of the full-length transcript variant for neurokinin-1 receptor in human whole blood associated with enhanced reinforcement of clot by substance-P

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Abstract

We have recently reported that a neurotransmitter for pain, substance-P (SP), promotes platelet-dependent clot formation through neurokinin-1 receptors (NK1Rs), in which leukocytes appear to be involved (J Thromb Thrombolysis 2009;27:280–6). Two naturally occurring splice isoforms of NK1R with different signal transduction potency, namely the full-length and the truncated NK1Rs are identified. It is known that human leukocytes express truncated NK1Rs, while in vivo expression of the full-length NK1R has not yet been fully clarified. Modulatory effects of alternative splicing for NK1Rs on clot formation also remain to be evaluated. Expression of the transcript variant mRNA for NK1Rs in human whole blood (n = 20) was evaluated by real-time reverse transcription polymerase chain reaction (RT-PCR). A 15 min time series of the strength of clot, formed after reloading of calcium in citrated whole blood with or without SP (10 nM) and a NK1R antagonist Spantide (1 μM), was measured by using oscillating-probe viscoelastometry. The full-length transcript variant was detected in 5 samples among 20. SP significantly increased the clot strength while Spantide suppressed the SP-derived change. The extent of modulation by SP/NK1R pathway in a subgroup with expression of the full-length transcript variant was three times as potent as those in another subgroup without expression. We conclude that expression of the full-length transcript variant for NK1R can be detected in human whole blood and that such expression is associated with the enhanced reinforcement of clot by SP. Further study is required to nominate this mRNA as a biomarker for prothrombotic risks in painful conditions such as perioperative period.

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Abbreviations

ANOVA:

Analysis of variance

CCL5:

Chemokine (C–C motif) ligand 5

CD11b/CD18 (Mac-1):

Macrophage-1 antigen

cDNA:

Complementary deoxyribonucleic acid

Ct:

Threshold cycle

mRNA:

Messenger ribonucleic acid

NK1R:

Neurokinin-1 receptor

PSGL1:

P-selectin glycoprotein ligand 1

RT-PCR:

Reverse transcription polymerase chain reaction

SP:

Substance-P

t-PA:

Tissue-plasminogen activator

VTE:

Venous thromboembolism

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Acknowledgments

This study was supported in part by Grants-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology in Japan (No. 22791453, 22791454, and 22591748).

Conflict of interest

There are no financial or other potential conflict of interests.

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Authors and Affiliations

  1. Department of Anesthesiology, Faculty of Medicine, Saitama Medical University, Iruma-gun, Saitama, 350-0495, Japan

    Toshiharu Azma, Taishin Ito, Masanori Tsukamoto, Hiroshi Hoshijima & Hirosato Kikuchi

  2. Department of Anesthesiology, Hiroshima University Hospital, Hiroshima, Japan

    Yuki Sugimoto

  3. Department of Anesthesiology, Wakayama Medical University, Wakayama, Japan

    Hiroyuki Kinoshita

  4. Department of Anesthesiology, Hiroshima General Hospital, Hatsukaichi, Hiroshima, Japan

    Masakazu Nakao

Authors
  1. Toshiharu Azma
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  2. Yuki Sugimoto
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  3. Hiroyuki Kinoshita
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  4. Taishin Ito
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  5. Masanori Tsukamoto
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Correspondence to Toshiharu Azma.

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Azma, T., Sugimoto, Y., Kinoshita, H. et al. Detection of the full-length transcript variant for neurokinin-1 receptor in human whole blood associated with enhanced reinforcement of clot by substance-P. J Thromb Thrombolysis 33, 329–337 (2012). https://doi.org/10.1007/s11239-011-0650-1

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  • DOI: https://doi.org/10.1007/s11239-011-0650-1

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