Hög aktivitetsnivå / Nature skalar upp HoloMonitoruppmärksamheten 2

I förra veckan publicerade Natures edition Scientific Reports en komplex forskningsrapport som var ett samarbete mellan forskare från Lunds Universitet, Karolinska Institutet och kinesiska Guangzhou Medical University.

Rapporten som publicerades 29/8 är benämnd

Nexilin/NEXN controls actin polymerization in smooth muscle and is regulated by myocardin family coactivators and YAP

Abstract
Nexilin, encoded by the NEXN gene, is expressed in striated muscle and localizes to Z-discs, influencing mechanical stability. We examined Nexilin/NEXN in smooth muscle cells (SMCs), and addressed if Nexilin localizes to dense bodies and dense bands and whether it is regulated by actin-controlled coactivators from the MRTF (MYOCD, MKL1, MKL2) and YAP/TAZ (YAP1 and WWTR1) families. NEXN expression in SMCs was comparable to that in striated muscles. Immunofluorescence and immunoelectron microscopy suggested that Nexilin localizes to dense bodies and dense bands. Correlations at the mRNA level suggested that NEXN expression might be controlled by actin polymerization. Depolymerization of actin using Latrunculin B repressed the NEXN mRNA and protein in bladder and coronary artery SMCs. Overexpression and knockdown supported involvement of both YAP/TAZ and MRTFs in the transcriptional control of NEXN. YAP/TAZ and MRTFs appeared equally important in bladder SMCs, whereas MRTFs dominated in vascular SMCs. Expression of NEXN was moreover reduced in situations of SMC phenotypic modulation in vivo. The proximal promoter of NEXN conferred control by MRTF-A/MKL1 and MYOCD. NEXN silencing reduced actin polymerization and cell migration, as well as SMC marker expression. NEXN targeting by actin-controlled coactivators thus amplifies SMC differentiation through the actin cytoskeleton, probably via dense bodies and dense bands.

Ur den klipper jag in utdrag som är PHI-relevant.

Material and Methods

Cell migration

Cell migration was measured using a scratch assay, and the cell confluence as well as the speed of the cell motility in the wound were followed over time using a real-time cell imaging system (HoloMonitor M4, Phase Holographic Imaging, Lund Sweden).
HBSMCs were seeded in 6-well plates (Sarstedt, 83.3920.005). 24 h after seeding, cells were transduced with 300 MOI of Ad-GFP-U6-h-NEXN-shRNA or control virus, respectively.
After another 24 h, the virus-containing media were exchanged for fresh media with 10% FBS to establish cell confluence. 12 h before the assay started, FBS was omitted from the medium. A scratch was made using a Gelloader Pipette Tip (Sarstedt, 70.1190.100), and the cell-free area was imaged every 40 min to monitor cell motility, cell divisions, and cell confluence.
Images were automatically acquired at 40 min intervals for the duration of the experiment (22 h) and stored by the HoloMonitor software system. For NEXN silencing using siRNA, HBSMCs were seeded in 6-well plates as above. 24 h after seeding, cells were transfected with 50 nM NEXN siRNA and 50 nM Negative Control siRNA, respectively. Cells were then incubated in the siRNA transfection medium for 24 h, followed by a 24 h incubation in DMEM/Ham’s F-12 medium with 10% FBS. 48 h after the first siRNA transfection, another siRNA transfection was done using DMEM/Ham’s F-12 medium with 0% FBS. 24 h after the second siRNA transfection a scratch was made, and images were acquired at 20 min intervals for the duration of the experiment (15 h) in the HoloMonitor as described above. Cell confluence and motility was analyzed using Hstudio M4.

Min kommentar
Denna forskningsrapport är som jag skrev i ingressen komplex. Eller rättare sagt komplicerad och riktigt svår att uttolka på ett begripligt sätt.
Hursom,av det jag förstår berör studien en gen ( Nexilin ) och dess påverkan på cellers muskelfunktion (häpp). Man har fokuserat på hjärt-kärlsystemet, mag-tarmkanalen och urinblåsan.
Genen nexilin är sen tidigare forskning känd för sin stabiliserande verkan, uttryckt som mechanical stability.
Då som del i muskelceller. Det forskarna nu studerat är hur genen fungerar i annat sammanhang.
 Alltså hjärt-kärl,mag-tarm och urinblåsa och deras uppbyggnad på cellnivå.
Resultaten de kommit fram till är som jag förstår det en utökad kunskap om genen och dess funktion.
Som sagt, en komplex forskning. Förmodligen söker de svar på nånting så svårt och komplicerat som kroppens DNA-kedja och dess minsta beståndsdelar. Eftersom forskningen bedrevs på 3 fronter kan man anta att den har stort intresse bland DNA forskare världen över.
I det sammanhanget är det absolut värdefullt för PHI,s vidkommande att deras HoloMonitorteknik visat sig vara användbar. Sen att få denna forskning,där PHI och HoloMonitorn omnämns, publicerad i det högst rankade organet bland forskare och vetenskapsmän (Nature) är naturligtvis en kvalitetsstämpel av rang.
Bolaget har ännu inte uppmärksammat denna rapport + föregående Natureinlägg på sin hemsida.
Jag tror att de kommer sätta bägge rapporterna inom guldram och stolt visa upp dessa för de initierade.