Serotonin-2 (5-HT2) Receptor-Mediated Signal Transduction in Human Ciliary Muscle Cells: Role in Ocular Hypotension

2006 ◽  
Vol 22 (6) ◽  
pp. 389-401 ◽  
Author(s):  
Najam A. Sharif ◽  
Curtis R. Kelly ◽  
Julie Y. Crider ◽  
Terry L. Davis
Keyword(s):  
Signal Transduction ◽  
Muscle Cells ◽  
Ciliary Muscle ◽  
Ocular Hypotension

Effect of Latanoprost Acid and Pilocarpine on Cultured Rabbit Ciliary Muscle Cells

2007 ◽  
Vol 39 (4) ◽  
pp. 232-240 ◽  
Author(s):  
Yisheng Zhong ◽  
Jian Gao ◽  
Wen Ye ◽  
Ping Huang ◽  
Yu Cheng ◽  
...  
Keyword(s):  
Muscle Cells ◽  
Ciliary Muscle

Abstract 486: Artificial miRNA Based Adenoviral Adam17 Silencing Reveals Essential Role of This Metalloprotease For EGF Receptor Transactivation In Primary Cultured Vascular Smooth Muscle Cells

Hypertension ◽  
2012 ◽  
Vol 60 (suppl_1) ◽  
Author(s):  
Katherine Elliott ◽  
Allison Bourne ◽  
Takehiko Takayanagi ◽  
Akira Takaguri ◽  
Kunie Eguchi ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Gene Silencing ◽  
Vascular Smooth Muscle Cells ◽  
Egf Receptor ◽  
Muscle Cells ◽  
Vascular Cells ◽  
Receptor Transactivation

siRNA mediated gene silencing has been recently utilized as a powerful molecular tool to study the functional significance of a specific protein. However, due to the transient nature of silencing and insufficient transfection efficiency, this approach can be problematic in primary cell culture. To overcome such weakness of the siRNA based silencing and in order to establish reliable gene silencing in vascular cells, we devised an adenoviral-encoded miRNA based gene silencing system. Here we report the results of silencing ADAM17 in cultured rat vascular smooth muscle cells (VSMCs) and its functional consequences in angiotensin II (AngII) signal transduction. Four distinct miRNA sequences targeting rat ADAM17 were chosen based on recommendations from Invitrogen’s Block-iT RNAi Designer algorithm. The miRNA sequences were inserted into a mammalian expression vector, pcDNA 6.2-GW/EmGFP-miR, and the effective silencing by these vectors was confirmed in HEK cells expressing HA-tagged rat ADAM17. The 4 cassettes carrying the miRNAs were inserted into pAd/CMV/V5-DEST and adenoviral solutions were obtained. Greater than 95% silencing of ADAM17 was achieved when VSMC were infected with 100-200 moi of the ADAM17 miRNA encoding adeonvirus for 72 h with enhancement of infection by fugene6. Relatively linear time and concentration dependencies were observed between 1 to 3 days and 10 to 100 moi of the infection. A miR-ADAM17 (100 moi) but not miR-control (100 moi) completely inhibited 100 nM AngII-induced HB-EGF shedding in VSMCs as assessed by a reporter assay. A miR-ADAM17 but not miR-control also inhibited AngII-induced EGF receptor transactivation and subsequent ERK1/2 activation in VSMCs as assessed by immunoblotting with phospho-selective antibodies. In conclusion, ADAM17 was found to be a major sheddase for HB-EGF contributing to the growth promoting signals induced by AngII in VSMCs. An artificial miRNA-base adenoviral approach appears to be a reliable gene-silencing strategy for signal transduction research in primary cultured vascular cells.

Signal Transduction Pathways in Smooth Muscle Cells

2004 ◽  
pp. 213-231 ◽  
Author(s):  
Sabina Vogel ◽  
Thomas Kubin ◽  
Miroslav Barancik ◽  
Elisabeth Deindl ◽  
Dietmar von der Ahe ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Signal Transduction Pathways

Functional organization of ciliary muscle in the cat

1962 ◽  
Vol 203 (5) ◽  
pp. 857-859 ◽  
Author(s):  
Harris Ripps ◽  
Irwin M. Siegel ◽  
William B. Getz
Keyword(s):  
Functional Organization ◽  
Muscle Cells ◽  
Nerve Fibers ◽  
Fixed Number ◽  
Ciliary Muscle ◽  
Frequency Variation ◽  
Efferent Nerve ◽  
Ocular Accommodation ◽  
Before And After ◽  
Stimulation Of

Ocular accommodation was elicited by electric stimulation of the ciliary ganglion in cats. The responses to frequency variation were recorded before and after cutting a branch of the postganglionic motor supply to the ciliary muscle. At all stimulus frequencies the responses obtained after section were shown to be a constant fraction of those obtained in the intact preparation. This suggests that a fixed number of muscle cells are affected when a portion of the efferent nerve fibers is cut. It was concluded that smooth ciliary muscle is organized as independent functional units, akin to the motor-unit arrangement of skeletal muscle; a given nerve fiber influences the response of a limited number of muscle cells.

β-Adrenoceptor activation and PKA regulate delayed rectifier K+ channels of vascular smooth muscle cells

1998 ◽  
Vol 275 (2) ◽  
pp. H448-H459 ◽  
Author(s):  
E. Alejandro Aiello ◽  
A. Todd Malcolm ◽  
Michael P. Walsh ◽  
William C. Cole
Keyword(s):  
Signal Transduction ◽  
Smooth Muscle ◽  
Portal Vein ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Catalytic Subunit ◽  
Delayed Rectifier ◽  
Rabbit Portal Vein

Macroscopic 4-aminopyridine (4-AP)-sensitive, delayed rectifier K+ current of vascular smooth muscle cells is increased during β-adrenoceptor activation with isoproterenol via a signal transduction pathway involving adenylyl cyclase and cAMP-dependent protein kinase (PKA) (Aiello, E. A., M. P. Walsh, and W. C. Cole. Am. J. Physiol. 268 ( Heart Circ. Physiol. 37): H926–H934, 1995.). In this study, we identified the single delayed rectifier K+(KDR) channel(s) of rabbit portal vein myocytes affected by treatment with isoproterenol or the catalytic subunit of PKA. 4-AP-sensitive KDR channels of 15.3 ± 0.6 pS ( n = 5) and 14.8 ± 0.6 pS ( n = 5) conductance, respectively, were observed in inside-out (I-O) and cell-attached (C-A) membrane patches in symmetrical KCl recording conditions. The kinetics of activation (time constant of 10.7 ± 3.02 ms) and inactivation (fast and slow time constants of 0.3 and 2.5 s, respectively) of ensemble currents produced by these channels mimicked those reported for inactivating, 4-AP-sensitive whole cell KDR current of vascular myocytes. Under control conditions, the open probability ( NP o) of KDR channels of C-A membrane patches at −40 mV was 0.014 ± 0.005 ( n = 8). Treatment with 1 μM isoproterenol caused a significant, approximately threefold increase in NP o to 0.041 ± 0.02 ( P < 0.05). KDR channels of I-O patches exhibited rundown after ∼5 min, which was not affected by ATP (5 mM) in the bath solution. Treatment with the purified catalytic subunit of PKA (50 nM; 5 mM ATP) restored KDRchannel activity and caused NP o to increase from 0.011 ± 0.003 to 0.138 ± 0.03 ( P < 0.05; n = 11). These data indicate that small-conductance, 15-pS KDRchannels are responsible for inactivating the macroscopic delayed rectifier K+ current of rabbit portal vein myocytes and that the activity of these channels is enhanced by a signal transduction mechanism involving β-adrenoceptors and phosphorylation by PKA at a membrane potential consistent with that observed in the myocytes in situ.

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