Chromatin remodeler Chd7 regulates photoreceptor development and outer segment length.

Mutations in the chromatin remodeling factor CHD7 are the predominant cause of CHARGE syndrome, a congenital disorder that frequently includes ocular coloboma. Although CHD7 is known to be required for proper ocular morphogenesis, its role in retinal development has not been thoroughly investigated. Given that individuals with CHARGE syndrome can experience visual impairment even in the absence of coloboma, a better understanding of CHD7 function in the retina is needed. In this study, we characterized the expression pattern of Chd7 in the developing zebrafish and mouse retina and documented ocular and retinal phenotypes in Chd7 loss-of-function mutants. Zebrafish Chd7 was expressed throughout the retinal neuroepithelium when retinal progenitor cells were actively proliferating, and later in subsets of newly post-mitotic retinal cells. At stages of retinal development when most retinal cell types had terminally differentiated, Chd7 expression remained strong in the ganglion cell layer and in some cells in the inner nuclear layer. Intriguingly, strong expression of Chd7 was also observed in the outer nuclear layer where it was co-expressed with markers of post-mitotic cone and rod photoreceptors. Expression of mouse CHD7 displayed a similar pattern, including expression in the ganglion cells, subsets of inner nuclear layer cells, and in the distal outer nuclear layer as late as P15. Two different mutant chd7 zebrafish lines were characterized for ocular and retinal defects. These mutants displayed microphthalmia, reduced numbers of cone photoreceptors, and truncated rod and cone photoreceptor outer segments. Reduced cone photoreceptor number and abnormal outer segments were also observed in heterozygous Chd7 mutant mice. Taken together, our results in zebrafish and mouse reveal a conserved, previously undescribed role for Chd7 in retinal development and photoreceptor outer segment morphogenesis. Moreover, our work suggests an avenue of future investigation into the pathogenesis of visual system defects in CHARGE syndrome.

Validation of a laser-assisted wound measurement device in a wound healing model.

In the treatment and monitoring of a diabetic or chronic wound, accurate and repeatable measurement of the wound provides indispensable data for the patient's medical record. This study aims to measure the accuracy of the laser-assisted wound measurement (LAWM) device against traditional methods in the measurement of area, depth and volume. We measured four 'healing' wounds in a Play-Doh(®) -based model over five subsequent states of wound healing progression in which the model was irregularly filled in to replicate the healing process. We evaluated the LAWM device against traditional methods including digital photograph assessment with National Institutes of Health ImageJ software, measurements of depth with a ruler and weight-to-volume assessment with dental paste. Statistical analyses included analysis of variance (ANOVA) and paired t-tests. We demonstrate that there are significantly different and nearly statistically significant differences between traditional ruler depth measurement and LAWM device measurement, but there are no statistically significant differences in area measurement. Volume measurements were found to be significantly different in two of the wounds. Rate of percentage change was analysed for volume and depth in the wound healing model, and the LAWM device was not significantly different than the traditional measurement technique. While occasionally inaccurate in its absolute measurement, the LAWM device is a useful tool in the clinician's arsenal as it reliably measures rate of percentage change in depth and volume and offers a potentially aseptic alternative to traditional measurement techniques.

Pilot study to evaluate a novel three-dimensional wound measurement device.

As the burden of diabetes continues to grow and treatment standards require careful tracking of wound progress, clinicians increasingly need to rely on technological improvements in wound measurement technologies to track the progress of their treatments. This study aims to determine the accuracy of a new three-dimensional wound measurement (3DWM) device against laser-assisted wound measurement (LAWM) devices and traditional methods of wound measurement. Using several wound models, we demonstrate that the 3DWM device measures wound area, depth and volume similarly to the other methods tested. This is especially apparent when changes in wound measurements were compared between the two devices. Differences between the two technologies were apparent when analysing wound measurement time and measurement repeatability. There was a significantly lower incidence of error in measurements between the 3DWM device and the LAWM device. Finally, the measurement time was significantly faster with the 3DWM device compared to the LAWM device. Together, these data demonstrate that the 3DWM device provides an accurate and reproducible method for measuring changes in wound healing similar to other available technologies. Further, the use of the 3DWM device provides a faster and more consistent measurement, which is critical for clinical application and use.

Her9/Hes4 is required for retinal photoreceptor development, maintenance, and survival.

The intrinsic and extrinsic factors that regulate vertebrate photoreceptor specification and differentiation are complex, and our understanding of all the players is far from complete. Her9, the zebrafish ortholog of human HES4, is a basic helix-loop-helix-orange transcriptional repressor that regulates neurogenesis in several developmental contexts. We have previously shown that her9 is upregulated during chronic rod photoreceptor degeneration and regeneration in adult zebrafish, but little is known about the role of her9 during retinal development. To better understand the function of Her9 in the retina, we generated zebrafish her9 CRISPR mutants. Her9 homozygous mutants displayed striking retinal phenotypes, including decreased numbers of rods and red/green cones, whereas blue and UV cones were relatively unaffected. The reduction in rods and red/green cones correlated with defects in photoreceptor subtype lineage specification. The remaining rods and double cones displayed abnormal outer segments, and elevated levels of apoptosis. In addition to the photoreceptor defects, her9 mutants also possessed a reduced proliferative ciliary marginal zone, and decreased and disorganized Müller glia. Mutation of her9 was larval lethal, with no mutants surviving past 13 days post fertilization. Our results reveal a previously undescribed role for Her9/Hes4 in photoreceptor differentiation, maintenance, and survival.

Author Correction: Her9/Hes4 is required for retinal photoreceptor development, maintenance, and survival.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

The role of estrogen in the modulation of autologous fat graft outcomes.

BACKGROUND: Autologous fat grafting is a widely used procedure, yet the mechanisms that regulate graft outcomes are poorly understood. Estrogen signaling is a potent regulator of lipid handling, inflammation, fibrosis, and adipocyte progenitor recruitment in adipose tissues. To date, no studies have investigated the effect of circulating estrogens on fat graft outcomes. METHODS: Immunosuppressed (Nu/Nu) mice underwent ovariectomy or sham surgery. Forty-five days later, half the mice (donors) were killed, and adipose tissue was taken and transplanted into the remaining cohort (recipients). Forty-five days after transplantation, grafts were dissected, weighed, and assessed for expression of vascular endothelial growth factor, estrogen receptor-α, and vascular density. RESULTS: Grafts harvested from and transplanted into sham environments are smaller but more highly vascularized compared with ovariectomy environments. The estrogenic effects on grafts are more critical at the site of the donor tissue than the recipient. Finally, expression of estrogen receptor-α in the grafted tissue correlates with the observed graft characteristics, which is altered by both the donor and recipient environments. CONCLUSIONS: Circulating estrogens have significant effects on fat graft outcomes, primarily at the site of the donor tissue. As there are well-established depot-specific estrogenic responses, the choice of adipose depot used as a donor for fat grafting may affect outcomes. In addition, outcomes may be confounded by the patient's hormonal status. Understanding the mechanisms by which estrogen signaling regulates graft outcomes is important in refining this commonly used clinical procedure.

Validation of a laser-assisted wound measurement device for measuring wound volume.

BACKGROUND: Accurate and precise wound measurement is an essential part of the medical record when monitoring a patient with a chronic wound. This study was designed to determine if a new device, a laser-assisted wound measurement (LAWM) device, provides valid measurements for wound area, depth, and volume. METHODS: We compared four methods to evaluate the area and volume of 12 wounds of differing size and depth that were created on the dorsum of a sacrificed pig. We evaluated the LAWM device, digital photograph assessment with National Institutes of Health ImageJ software, measurements of depth with a ruler, and weight-to-volume assessment with dental paste. We then sought to cross validate this data with further analyses obtained from these measurements using a Play-Doh®-based wound as a model for constant area with different depths. RESULTS: We demonstrate that the LAWM device measures wound area accurately. Depth (and therefore volume) measurements, however, are artificially low. This inaccuracy is the same for shallow and deep wounds. CONCLUSIONS: The inaccuracy in the depth and volume measurements with the LAWM device results in an artificially low measurement. However, this may not affect percentage difference measurements. Further studies will need to be performed to determine if this device can accurately determine wound changes in the clinical setting.