hypertonic buffer  (Gold Biotechnology Inc)

Journal: Frontiers in Medicine

Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1302119

Figure Lengend Snippet: (A–C) Cyst severity is significantly reduced in vector-treated and hypertonic buffer-injected eyes at 1 month post injection (MPI). OCT images of vector-treated, hypertonic buffer-injected, and untreated eyes were collected at 1, 2, and 5 MPI. Volumetric scans were taken centering on the optic nerve. Representative OCT images demonstrate cyst changes at 1 MPI (A) . Cyst severity (B) was determined via manual segmentation of cyst area using the ImageJ software. Multiple comparisons were performed between every group at each time point. Non-significant comparisons were not annotated in the graphs. Vector-treated ( n = 10) or hypertonic buffer-injected Rs1 -KO eyes demonstrated significantly less cyst severity than untreated eyes (B) at 1 MPI. ONL thickness (C) remained stable from 1 to 5 MPI in all cohorts; however, at 5 MPI, vector treated eyes had statistically thinner ONLs than untreated eyes.

Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

Techniques: Plasmid Preparation, Injection, Software

Journal: Frontiers in Medicine

Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1302119

Figure Lengend Snippet: (A–E) Hypertonic buffer robustly outperforms untreated and vector-treated eyes in light-adapted ERGs. Cone-dependent retina function was measured using two separate light-adapted ERG assays—the 5 Hz flicker (A) and 3.0 flash (B) . ERG amplitudes are shown from 1-month post injection (MPI) to 5 MPI for vector-treated, hypertonic buffer-injected, and untreated eyes in Rs1 -KO mice, and representative ERG traces at 5 MPI are illustrated. In response to both light-adapted assays, hypertonic buffer-injected eyes showed higher amplitudes than untreated and vector-treated eyes to 5 MPI (A) . The combined rod-cone function was measured by the 3.0 cd·s/m 2 standard combined response (SCR) under dark-adapted conditions (C) . At 1, 3, and 5-MPI, there was no difference between vector-treated, hypertonic buffer-injected, or untreated eyes. Isolated rod function was measured using 0.01 cd·s/m 2 dim flash after dark adaptation (D) . There was no persistent significant difference between hypertonic buffer-injected, isotonic buffer-injected, or untreated eyes in rod dominant ERG settings.

Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

Techniques: Plasmid Preparation, Injection, Isolation

Journal: Frontiers in Medicine

Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1302119

Figure Lengend Snippet: (A,B) Vector-treated and hypertonic buffer-injected Rs1 -KO mice perform faster in a light-adapted swim maze than untreated Rs1 -KO mice. Mice were placed in a swim maze and testing was performed under light-adapted (A) or dark-adapted (B) conditions. Time-to-platform was recorded. Each data point is representative of the average time-to-platform for a single mouse. In light-adapted conditions (untreated n = 8, vector-treated n = 4, hypertonic buffer n = 10.), we observed that vector-treated mice (2.92 s) and hypertonic buffer-injected mice (3.05 s) had a generally faster trend in average swim performance when finding a random platform than untreated mice (4.50 s) in lighted environments. In a dark-adapted environment, there is a less impactful effect on average time-to-platform between vector-treated (11.31 s), hypertonic buffer-injected (13.78 s), and untreated eyes (11.13 s), suggesting a limited effect on visual function in the dark (B) .

Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

Techniques: Plasmid Preparation, Injection

Journal: Frontiers in Medicine

Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1302119

Figure Lengend Snippet: (A–C) Cyst area is reduced in hypertonic buffer and isotonic buffer-injected eyes, with greater reduction observed in the hypertonic buffer group. OCT images were collected at 2- and 3-weeks post injection (WPI), 1-month post-injection (MPI), and 2 MPI. Cyst area quantification (A) and outer nuclear layer (ONL) thickness (B) measurements were taken to assess cyst severity and photoreceptor cell survival, respectively. Representative OCT images demonstrate cysts at respective time points in all experimental groups (C) . Each data point is representative of one eye. At 3 WPI, there was less cyst severity in the hypertonic buffer-injected eyes compared to untreated eyes (A) . At 1 MPI, hypertonic and isotonic buffer-treated eyes both have less severe cysts than untreated and sham punctured eyes. This effect persisted until 2 MPI for the hypertonic buffer-injected eyes compared to untreated and sham punctured eyes. ONL thickness remains stable to 5 MPI and does not significantly differ between injection groups and untreated controls (B) .

Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

Techniques: Injection

Journal: Frontiers in Medicine

Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1302119

Figure Lengend Snippet: (A–E) Hypertonic buffer-injected eyes experience a robust rescue of cone functioning. Cone-dependent retina function was measured using two separate light-adapted ERG assays—the 3.0 cd·s/m 2 flash (A) , and the 5 Hz flicker (B) in hypertonic buffer, isotonic buffer, and sham punctured eyes in Rs1 -KO mice. Each data point represents one eye. ERG amplitudes are shown from 1-month post injection (MPI) to 5 MPI. In both light-adapted assays (A,B) , hypertonic and isotonic buffer-injected eyes showed higher amplitudes compared to untreated eyes at 1 MPI. Hypertonic buffer-injected eyes showed improvements over untreated and sham punctured eyes until 5 MPI. The combined rod-cone function was measured by the 3.0 cd·s/m 2 SCR under dark-adapted conditions (C) . Rod-dependent retina function was measured by the 0.01 dim flash (D) . In the 0.01 dim flash, hypertonic buffer-injected eyes showed improvements over untreated at 1 MPI and over both untreated and isotonic buffer-injected eyes at 2 MPI. However, significant differences over time were not observed.

Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

Techniques: Injection

Journal: Frontiers in Medicine

Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1302119

Figure Lengend Snippet: (A–F) Peak cyst severity on OCT is negatively associated with cone function on ERG at 5 MPI. Peak cyst severity of hypertonic buffer-injected, isotonic buffer-injected, sham-punctured and untreated eyes was determined by comparing the cyst area measurements of each eye at 1, 2, and 5 MPI. The cyst area of each eye representing their peak severity was correlated with its ERG amplitudes at 5 MPI. Each dot represents one eye. A negative correlation was found between cyst area and ERG amplitudes in the light-adapted 5 Hz flicker and 3.0 cd·s/m 2 flash (A,B) . Little to no correlation was found between peak cyst area and ERG amplitudes in dark-adapted 0.01 cd·s/m 2 or 3.0 cd·s/m 2 SCR (C–F) . These findings suggest increasing cyst area has greater impact on impairing cone electrical function, than rod photoreceptors.

Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

Techniques: Injection

Journal: Frontiers in Medicine

Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

doi: 10.3389/fmed.2024.1302119

Figure Lengend Snippet: (A,B) Increased cone density at 5 months after subretinal injections of a hypertonic or an isotonic buffer. Retinal sections were collected from untreated Rs1 -KO eyes, hypertonic buffer-injected eyes, isotonic buffer-injected eyes, and sham puncture eyes at approximately 6-months of age. Sections were stained by PNA to visualize cone outer segments. The number of cone outer segments per 100 μm of the retina was quantified by three participants masked to treatment groups, and quantifications by these three participants were averaged. Representative immunofluorescent staining (red: PNA) of cone outer segments (arrow), demonstrating increased density of cones in hypertonic buffer-injected and isotonic buffer-injected eyes compared to untreated eyes with sparse cone staining (A) . Synapses were visualized by an anti-RIBEYE antibody (green). Each data point represents the averaged cone density of one eye. Hypertonic buffer-injected eyes and isotonic buffer-injected eyes show significantly higher cone density per 100 μm than untreated Rs1 -KO eyes (B) .

Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

Techniques: Injection, Staining