scholarly journals Cyclic Adenosine Monophosphate, Inositol 1,4,5-trisphosphate, Calcium, and Phosphorylated Myosin Light Chain Regulation Through M2 and M3 Muscarinic Receptors of Scleral Fibroblast Cells in Rat Myopia Model

2021 ◽  
Vol 9 (B) ◽  
pp. 890-899
Author(s):  
Nanda Wahyu Anandita ◽  
Nurdiana Nurdiana ◽  
Endang Sri Wahyuni ◽  
Hidayat Sujuti
Keyword(s):  
Animal Models ◽  
Muscarinic Receptor ◽  
Light Chain ◽  
Rattus Norvegicus ◽  
Myosin Light Chain ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Fibroblast Cells ◽  
Inositol 1,4,5 Trisphosphate ◽  
M3 Muscarinic Receptor

AIM: This study aims to investigate the concentration of cyclic adenosine monophosphate (cAMP), inositol 1,4,5-trisphosphate (IP3), calcium (Ca2+), and the expression phosphorylated myosin light chain (MLC) in Rattus norvegicus scleral fibroblast cells. METHODOLOGY: This study utilized an in vitro experimental study by applying Rattus norvegicus scleral fibroblast cell culture. The cultured cells were divided into control and lens-induced myopia (LIM) groups. The control and LIM culture groups were each divided into five groups, namely, negative control, 0.1 μM acetylcholine, 0.1 μM himbacine, 0.1 μM methoctramine, and 0.1 μM 4-DAMP group. The cAMP, IP3, and Ca2+ concentration were analyzed in the 0th, 5th, 10th, 20th, and 30th. The phosphorylated MLC expression was analyzed using confocal microscope. RESULTS: In the LIM group, the highest cAMP concentration is visible at the 10th min on the himbacine group (0.304 ± 0; p = 0.043) and on the 4-DAMP group (0.346 ± 0; p = 0.043). The highest IP3 concentration is found on the LIM group at the 20th min in comparison to the control group (2503.6 ± 11 vs. 2039.2 ± 2.1; p = 0.046). The highest Ca2+ concentration belongs to the 4-DAMP treatment group from the 5th to the 30th min. The highest average phosphorylated MLC expression value in the LIM group is shown by the 0.1μM 4-DAMP treatment (184.2 ± 37.9c au). CONCLUSION: The regulation of cAMP, IP3, Ca2+, and phosphorylated MLC on the M2 and M3 muscarinic receptor of the scleral fibroblast cells of myopia animal models differs from normal animal models which may be due to interactions of M2 and M3 muscarinic receptor as compensation reaction or crosstalk on myopia induction.

A calmodulin antagonist reveals a calmodulin-independent interdomain interaction essential for activation of inositol 1,4,5-trisphosphate receptors

2008 ◽  
Vol 416 (2) ◽  
pp. 243-253 ◽  
Author(s):  
Yi Sun ◽  
Colin W. Taylor
Keyword(s):  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Channel Gating ◽  
Binding Motif ◽  
Calmodulin Antagonist ◽  
Binding Peptide ◽  
Selective Antagonist ◽  
Inositol 1,4,5 Trisphosphate ◽  
Interdomain Interaction

CaM (calmodulin) has been implicated in the regulation of IP3R [IP3 (inositol 1,4,5-trisphosphate) receptors] and a recent report suggested that CaM tightly tethered to IP3R was essential for IP3R activation [Nadif Kasri, Torok, Galione, Garnham, Callewaert, Missiaen, Parys and De Smedt (2006) J. Biol. Chem. 281, 8332–8338]. In the present study, we confirm that a CaM-binding peptide derived from MLCK (myosin light chain kinase) inhibits IP3-evoked Ca2+ release via all three IP3R subtypes. However, inhibition by MLCK peptide is not mimicked by other CaM antagonists that effectively block regulation of IP3R by CaM. Inhibition by MLCK peptide is rapid, fully reversible and occurs under conditions where there is no CaM associated with IP3R. MLCK peptide stimulates IP3 binding to IP3R1 and to its bacterially expressed N-terminal, but not after removal of the suppressor domain (residues 1–224). We suggest that MLCK peptide mimics a sequence within the suppressor domain that is similar to a 1-8-14 CaM-binding motif. The peptide may thereby unzip an interdomain interaction that is essential for IP3R activation. We conclude that CaM is not essential for IP3R activation, and that MLCK peptide is a selective antagonist of the IP3R that binds directly to the N-terminal to uncouple IP3 binding from channel gating. The results of the present study highlight the importance of the suppressor domain in IP3R activation and suggest that MLCK peptide may provide a route to novel non-competitive antagonists of IP3R.

676 Type 3 Muscarinic Receptor Deficiency Is Associated With Impaired Paracellular Permeability and Activation of Myosin Light Chain (MLC) 2

2013 ◽  
Vol 144 (5) ◽  
pp. S-123-S-124
Author(s):  
Leon McLean ◽  
Luigi Notari ◽  
Rex Sun ◽  
Jennifer A. Bohl ◽  
Shu Yan ◽  
...  
Keyword(s):  
Muscarinic Receptor ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Paracellular Permeability ◽  
Type 3

Phosphodiesterase 2A Localized in the Spinal Cord Contributes to Inflammatory Pain Processing

2014 ◽  
Vol 121 (2) ◽  
pp. 372-382 ◽  
Author(s):  
Wiebke Kallenborn-Gerhardt ◽  
Ruirui Lu ◽  
Aaron Bothe ◽  
Dominique Thomas ◽  
Jessica Schlaudraff ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Animal Models ◽  
Dorsal Horn ◽  
Cyclic Nucleotides ◽  
Inflammatory Pain ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Cyclic Guanosine Monophosphate ◽  
Guanosine Monophosphate ◽  
Pain Processing

Abstract Background: Phosphodiesterase 2A (PDE2A) is an evolutionarily conserved enzyme that catalyzes the degradation of the cyclic nucleotides, cyclic adenosine monophosphate, and/or cyclic guanosine monophosphate. Recent studies reported the expression of PDE2A in the dorsal horn of the spinal cord, pointing to a potential contribution to the processing of pain. However, the functions of PDE2A in spinal pain processing in vivo remained elusive. Methods: Immunohistochemistry, laser microdissection, and quantitative real-time reverse transcription polymerase chain reaction experiments were performed to characterize the localization and regulation of PDE2A protein and messenger RNA in the mouse spinal cord. Effects of the selective PDE2A inhibitor, BAY 60–7550 (Cayman Chemical, Ann Arbor, MI), in animal models of inflammatory pain (n = 6 to 10), neuropathic pain (n = 5 to 6), and after intrathecal injection of cyclic nucleotides (n = 6 to 8) were examined. Also, cyclic adenosine monophosphate and cyclic guanosine monophosphate levels in spinal cord tissues were measured by liquid chromatography tandem mass spectrometry. Results: The authors here demonstrate that PDE2A is distinctly expressed in neurons of the superficial dorsal horn of the spinal cord, and that its spinal expression is upregulated in response to hind paw inflammation. Administration of the selective PDE2A inhibitor, BAY 60–7550, increased the nociceptive behavior of mice in animal models of inflammatory pain. Moreover, BAY 60–7550 increased the pain hypersensitivity induced by intrathecal delivery of cyclic adenosine monophosphate, but not of cyclic guanosine monophosphate, and it increased the cyclic adenosine monophosphate levels in spinal cord tissues. Conclusion: Our findings indicate that PDE2A contributes to the processing of inflammatory pain in the spinal cord.

scholarly journals PENGARUH PEMBERIAN KOPI TERHADAP KUALITAS SPERMATOZOA TIKUS WISTAR JANTAN (Rattus norvegicus) YANG DIBERI PAPARAN ASAP ROKOK

2015 ◽  
Vol 3 (2) ◽  
Author(s):  
Ayu L. Dja’afara ◽  
Benny Wantouw ◽  
Lydia Tendean
Keyword(s):  
Cigarette Smoke ◽  
Rattus Norvegicus ◽  
Wistar Rats ◽  
Semen Quality ◽  
Sperm Quality ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Control Group ◽  
Camp Production

Abstract: Coffee contains caffeine which acts to increase cyclic adenosine monophosphate (cAMP) production in order to stimulate the motility of spermatozoa. Smoking affects the process of spermatogenesis, semen quality, and testosterone level. This study aimed to determine the effect of coffee on sperm quality of wistar rats exposed to cigarette smoke. This was a descriptive observational study. Samples were 6 wistar rats divided into 3 groups, each of 2 rats. The control group (P0) was exposed to cigarette smoke of 2 cigarettes/day. The P1 group was exposed to cigarette smoke (2 cigarettes/day) and was given 40 mg coffee solution; and the P2 group was exposed to cigarette smoke (2 cigarettes/day) and was given 80 mg coffee solution. The results showed that rats in P2 group showed increases and improvement in the spermatozoa concentration 70.9x106/ml, motility of spermatozoa category A by 55%, and morphologically normal spermatozoa by 55.5%. Conclusion: Coffee can improve the sperm quality of wistar rats Rattus norvegicus exposed to cigarette smoke.Keywords: cigarette, coffee, sperm qualityAbstrak: Kopi mengandung kafein yang berfungsi meningkatkan produksi siklik adenosin monofosfat (cAMP) yang merangsang gerakan spermatozoa. Rokok memengaruhi proses spermatogenesis, kualitas semen, dan kadar hormon testosteron. Penelitian ini bertujuan untuk mengetahui pengaruh kopi terhadap kualitas spermatozoa tikus wistar jantan yang diberi paparan asap rokok. Penelitian ini bersifat observasional deskriptif. Sampel sebanyak 6 ekor tikus wistar jantan: 2 ekor tikus wistar jantan sebagai kontrol (P0) yang hanya diberi paparan asap rokok 2 batang/hari; 2 ekor tikus wistar jantan diberi paparan asap rokok 2 batang/hari dan 40 mg larutan kopi (P1); dan 2 ekor tikus wistar jantan diberi paparan asap rokok 2 batang/hari dan 80 mg larutan kopi (P2). Hasil penelitian memperlihatkan pada kelompok P2 terjadi peningkatan konsentrasi spermatozoa sebesar 70,9x106/ml, peningkatan motilitas spermatozoa kategori A sebesar 55% dan morfologi normal spermatozoa sebesar 55,5%. Simpulan: Kopi dapat meningkatkan kualitas spermatozoa tikus wistar jantan Rattus norvegicus yang diberi paparan asap rokok.Kata kunci: rokok, kopi, kualitas spermatozoa

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