Real-TimeReflectance Confocal Microscopy, a Noninvasive Tool for in vivo Quantitative Evaluation of Comedolysis in the Rhino Mouse Model

K. Nakano; K. Kiyokane; C. Benvenuto-Andrade; S. González

doi:10.1159/000096169

Comparison between reflectance confocal microscopy and 2-photon microscopy in early detection of cutaneous radiation injury in a mouse model in vivo

Journal of Biophotonics ◽

10.1002/jbio.201700337 ◽

2018 ◽

Vol 11 (10) ◽

pp. e201700337 ◽

Cited By ~ 2

Author(s):

Won Hyuk Jang ◽

Soonjae Kwon ◽

Sehwan Shim ◽

Won-Suk Jang ◽

Jae Kyung Myung ◽

...

Keyword(s):

Confocal Microscopy ◽

Early Detection ◽

Mouse Model ◽

Radiation Injury ◽

Reflectance Confocal Microscopy

Download Full-text

Conditional Up-Regulation of SERCA2a Exacerbates RyR2-Dependent Ventricular and Atrial Arrhythmias

International Journal of Molecular Sciences ◽

10.3390/ijms21072535 ◽

2020 ◽

Vol 21 (7) ◽

pp. 2535

Author(s):

Bin Liu ◽

Qing Lou ◽

Heather Smith ◽

Florencia Velez-Cortes ◽

Wolfgang H. Dillmann ◽

...

Keyword(s):

Confocal Microscopy ◽

Mouse Model ◽

Transgenic Mice ◽

Cardiac Disease ◽

Ventricular Myocytes ◽

Atrial Arrhythmias ◽

Knock Out ◽

Disease Therapy ◽

Upregulated Genes

Ryanodine receptor 2 (RyR2) and SERCA2a are two major players in myocyte calcium (Ca) cycling that are modulated physiologically, affected by disease and thus considered to be potential targets for cardiac disease therapy. However, how RyR2 and SERCA2a influence each others’ activities, as well as the primary and secondary consequences of their combined manipulations remain controversial. In this study, we examined the effect of acute upregulation of SERCA2a on arrhythmogenesis by conditionally overexpressing SERCA2a in a mouse model featuring hyperactive RyR2s due to ablation of calsequestrin 2 (CASQ2). CASQ2 knock-out (KO) mice were crossbred with doxycycline (DOX)-inducible SERCA2a transgenic mice to generate KO-TG mice. In-vivo ECG studies have shown that induction of SERCA2a (DOX+) overexpression markedly exacerbated both ventricular and atrial arrhythmias in vivo, compared with uninduced KO-TG mice (DOX-). Consistent with that, confocal microscopy in both atrial and ventricular myocytes demonstrated that conditional upregulation of SERCA2a enhanced the rate of occurrence of diastolic Ca release events. Additionally, deep RNA sequencing identified 17 downregulated genes and 5 upregulated genes in DOX+ mice, among which Ppp1r13l, Clcn1, and Agt have previously been linked to arrhythmias. Our results suggest that conditional upregulation of SERCA2a exacerbates hyperactive RyR2-mediated arrhythmias by further elevating diastolic Ca release.

Download Full-text

Observation of Chronic Graft-Versus-Host Disease Mouse Model Cornea with In Vivo Confocal Microscopy

Diagnostics ◽

10.3390/diagnostics11081515 ◽

2021 ◽

Vol 11 (8) ◽

pp. 1515

Author(s):

Shota Shimizu ◽

Shinri Sato ◽

Hiroko Taniguchi ◽

Eisuke Shimizu ◽

Jingliang He ◽

...

Keyword(s):

Confocal Microscopy ◽

Mouse Model ◽

Graft Versus Host Disease ◽

Pathological Condition ◽

Inflammatory Cells ◽

In Vivo Confocal Microscopy ◽

Host Disease ◽

Epithelial Damage ◽

Graft Versus Host

Graft-versus-host disease (GVHD) is a major complication after hematopoietic stem cell transplantation (HSCT), and ocular GVHD can cause severe dry eye disease that can lead to visual impairment. Epithelial damage, vascular invasion, corneal fibrosis, and corneal perforation may occur in severe cases. It is generally accepted that inflammatory cells such as dendritic cells and T cells contribute to this pathological condition. However, it is still unknown what pathological condition occurs on the ocular surface after HSCT, and when. We therefore observed the dynamics of inflammatory cells in the cornea of chronic GVHD (cGVHD) model mice from 1 to 4 weeks after bone marrow transplantation (BMT) by in vivo confocal microscopy (IVCM) and considered the relationship with the pathophysiology of ocular GVHD (tear volume, corneal epithelial damage). In the allogeneic group, neovascularization occurred in all eyes at 1 week after BMT, although almost all vessels disappeared at 2 weeks after BMT. In addition, we revealed that infiltration of globular cells, and tortuosity and branching of nerves in the cornea occurred in both cGVHD mice and human cGVHD patients. Thus, we consider that cGVHD mouse model study by IVCM reproduces the state of ocular GVHD and may contribute to elucidating the pathological mechanism for ocular GVHD.

Download Full-text

Targeted therapy for the post-operative conjunctiva: SPARC silencing reduces collagen deposition

British Journal of Ophthalmology ◽

10.1136/bjophthalmol-2018-311937 ◽

2018 ◽

Vol 102 (10) ◽

pp. 1460-1470 ◽

Cited By ~ 1

Author(s):

Li Fong Seet ◽

Yang Fei Tan ◽

Li Zhen Toh ◽

Stephanie WL Chu ◽

Ying Shi Lee ◽

...

Keyword(s):

Targeted Therapy ◽

Confocal Microscopy ◽

Mouse Model ◽

Anterior Segment ◽

Annexin V ◽

Glaucoma Surgery ◽

Tissue Response ◽

Layer By Layer ◽

Glaucoma Filtration Surgery

BackgroundTo develop targeted antifibrotic therapy for glaucoma filtration surgery; this study determines the effectiveness of small interfering RNA (siRNA) to reduce in vivo secreted protein acidic and rich in cysteine (SPARC) expression using the mouse model of conjunctival scarring.Methods Experimental surgery was performed as described for the mouse model of conjunctival scarring. Scrambled (siScram) or Sparc (siSparc) siRNAs, loaded on layer-by-layer (LbL) nanoparticles, were injected into the conjunctiva immediately after surgery. Expression of Sparc, Col1a1, Fn1 and Mmp14 was measured by real-time PCR and immunoblotting on days 7 and 14 postsurgery. Live imaging of the operated eyes was performed using slit lamp, anterior segment-optical coherence tomography and confocal microscopy. Tissue pathology was evaluated by histochemical and immunofluorescent analyses of operated conjunctival cryosections. Tissue apoptosis was quantitated by annexin V assay.Results siSparc, delivered via expanded LbL nanoparticles, significantly inhibited Sparc transcription in both day 7 (2.04-fold) and day 14 (1.39-fold) treated tissues. Sparc suppression on day 7 was associated with a significant reduction of Col1a1 (2.52-fold), Fn1 (2.89-fold) and Mmp14 (2.23-fold) mRNAs. At the protein level, both SPARC and collagen 1A1 (COL1A1) were significantly reduced at both time points with siSparc treatment. Nanoparticles were visualised within cell-like structures by confocal microscopy, while overt tissue response or apoptosis was not observed.Conclusions SPARC targeted therapy effectively reduced both SPARC and collagen production in the operated mouse conjunctiva. This proof-of-concept study suggests that targeted treatment of fibrosis in glaucoma surgery is safe and feasible, with the potential to extend to a range of potential genes associated with fibrosis.

Download Full-text