An expanded GCaMP reporter toolkit for functional imaging in Caenorhabditis elegans.

In living organisms, changes in calcium flux are integral to many different cellular functions and are especially critical for the activity of neurons and myocytes. Genetically encoded calcium indicators (GECIs) have been popular tools for reporting changes in calcium levels in vivo. In particular, GCaMPs, derived from GFP, are the most widely used GECIs and have become an invaluable toolkit for neurophysiological studies. Recently, new variants of GCaMP, which offer a greater variety of temporal dynamics and improved brightness, have been developed. However, these variants are not readily available to the Caenorhabditis elegans research community. This work reports a set of GCaMP6 and jGCaMP7 reporters optimized for C. elegans studies. Our toolkit provides reporters with improved dynamic range, varied kinetics, and targeted subcellular localizations. Besides optimized routine uses, this set of reporters is also well suited for studies requiring fast imaging speeds and low magnification or low-cost platforms.

An expanded GCaMP reporter toolkit for functional imaging in C. elegans.

In living organisms, changes in calcium flux are integral to many different cellular functions and are especially critical for the activity of neurons and myocytes. Genetically encoded calcium indicators (GECIs) have been popular tools for reporting changes in calcium levels in vivo . In particular, GCaMP, derived from GFP, are the most widely used GECIs and have become an invaluable toolkit for neurophysiological studies. Recently, new variants of GCaMP, which offer a greater variety of temporal dynamics and improved brightness, have been developed. However, these variants are not readily available to the Caenorhabditis elegans research community. This work reports a set of GCaMP6 and jGCaMP7 reporters optimized for C. elegans studies. Our toolkit provides reporters with improved dynamic range, varied kinetics, and targeted subcellular localizations. Besides optimized routine uses, this set of reporters are also well-suited for studies requiring fast imaging speeds and low magnification or low-cost platforms.

Metabolic, cardiopulmonary, and gait profiles of recently injured and noninjured runners.

OBJECTIVE: To examine whether runners recovering from a lower body musculoskeletal injury have different metabolic, cardiopulmonary, and gait responses compared with healthy runners. DESIGN: Cross-sectional study. SETTING: Research laboratory at an academic institution. METHODS: Healthy runners (n = 50) were compared with runners who were recently injured but had returned to running (n = 50). Both groups were participating in similar cross-training modalities such as swimming, weight training, biking, and yoga. Running gait was analyzed on a treadmill using 3-dimensional motion capture, and metabolic and cardiopulmonary measures were captured simultaneously with a portable metabolic analyzer. MAIN OUTCOME MEASURES: Rate of oxygen consumption, heart rate, ventilation, carbohydrate and fat oxidation values, gait temporospatial parameters and range of motion measures (ROM) in the sagittal plane, energy expenditure, and vertical displacement of the body's center of gravity (COG). RESULTS: The self-selected running speed was different between the injured and healthy runners (9.7 ± 1.1 km/h and 10.6 ± 1.1 km/h, respectively; P = .038). No significant group differences were noted in any metabolic or cardiopulmonary variable while running at the self-selected or standard speed (13.6 km/h). The vertical displacement of the COG was less in the injured group (8.4 ± 1.4 cm and 8.9 ± 1.4, respectively; P = .044). ROM about the right ankle in the sagittal plane at the self-selected running speed during the gait cycle was less in the injured runners compared with the healthy runners (P < .05). CONCLUSIONS: Runners with a recent lower body injury who have returned to running have similar cardiopulmonary and metabolic responses to running as healthy runners at the self-selected and standard speeds; this finding may be due in part to participation in cross-training modes that preserve cardiopulmonary and metabolic adaptations. Injured runners may conserve motion by minimizing COG displacement and ankle joint ROM during a gait cycle.