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The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats

Tasks that accurately measure dexterity in animal models are critical to understand hand function. Current rat behavioral tasks that measure dexterity ...

Burke Medical Research Institute

Chicks are bipedal precocious vertebrates that achieve adaptive locomotor skill within hours after hatching. Development of limb movement has been extensively studied in the chicken embryo, but few studies have focused on the preparations leading to precocious locomotor skill. Chicks typically hatch after 21 days of incubation, and recent studies provided evidence that the neural circuits for intralimb control of stepping are established between embryonic days (E) 18-20. It has also been shown that variations in light exposure during embryogenesis can accelerate or delay the onset of hatching and walking by 1 to 2 days. Our earlier work revealed that despite these differences in time to hatch, chicks incubated in different light conditions achieved similar locomotor skill on the day of hatching. Results suggested to us that light exposure during incubation may have accelerated development of locomotor circuits in register with earlier hatching. Thus, in this study, embryos were incubated in 1 of 3 light conditions to determine if development of interlimb coordination at a common time point, 19 days of incubation, varied with light exposure during embryogenesis. Leg muscle activity was recorded bilaterally and burst analyses were performed for sequences of spontaneous locomotor-related activity in one or more ankle muscles to quantify the extent of interlimb coordination in ovo. We report findings indicating that the extent of interlimb coordination varied with light exposure, and left-right alternating steps were a more reliable attribute of interlimb coordination for embryos incubated in constant bright light. We provide evidence that morphological development of the leg varied with light exposure. Based on these findings, we propose that light can accelerate the development of interlimb coordination in register with earlier hatching. Our results lead us to further propose that alternating left-right stepping is the default pattern of interlimb coordination produced by locomotor circuits during embryogenesis.

Light accelerates morphogenesis and acquisition of interlimb stepping in chick embryos.

This chapter discusses atomic force microscopy (AFM) for the benefit of microbiologists who are interested in using this technique to examine the structures and dynamics of bacteria. AFM is a powerful technique for imaging biological samples at the nanometer to micrometer scale under nondestructive conditions. In order to be imaged with AFM, bacteria must be supported by a surface, which presents challenges because many laboratory strains of bacteria are planktonic. Still, in nature many bacteria live at surfaces and interfaces. This chapter discusses the benefits and difficulties of different methods that have been used to support bacteria on surfaces for AFM imaging and presents two methods in detail used to successfully grow and image bacteria at solid-liquid and solid-air interfaces. Using these methods it is possible to study bacterial morphology and interactions in a native state. These explorations by AFM have important applications to the study of different kinds of bacteria, interfacial bacterial communities, and biofilms.

Atomic force microscopy of bacterial communities.

A new method of isolating host-independent Bdellovibrio bacteriovorus has been developed. Filtered suspensions of host-dependent cells are dropped in small volumes onto 0.2 microm membranes laid on rich media agar. Significant growth is observed within 1-2 days; these cells were confirmed to be B. bacteriovorus using microscopic observations and PCR.

Rapid isolation of host-independent Bdellovibrio bacteriovorus.

Domestic chicks walk within 3-4 hr after hatching following 21 days of incubation. However, differences in light exposure can vary incubation duration. Based on pilot studies, we predicted that there would be a positive relationship between incubation duration and locomotor competence at hatching. Embryos were incubated in one of three conditions that varied light duration and intensity, and overground locomotor performance was tested on the day of hatching. Chicks incubated in continuous bright light hatched 1-2 days earlier than chicks incubated in less or no light. Kinematic findings indicated that locomotor skill was similar across incubation conditions and led us to reject our hypothesis. We propose that light may accelerate locomotor development without adversely affecting skill. Our findings raise two important implications for future studies: whether light exposure accelerates locomotor circuit development; and/or it unmasks adaptive motor skill by accelerating development of other physiological systems.

Kinematic analysis of overground locomotion in chicks incubated under different light conditions.

In an earlier study of newly hatched chicks we reported that continuous bright light exposure throughout incubation accelerated locomotor development and continuous dark exposure delayed it, compared to less intense, intermittent light exposure. Commonly studied gait parameters indicated locomotor skill was similar across groups. However, dark incubated chicks walked with a greater step width, raising the possibility of differences in dynamic balance and control of forward progression. In this study, we established methods to retrospectively examine the previously published locomotor data for differences in lateral drift. We hypothesized that chicks incubated in darkness would exhibit more drift than chicks incubated in light. Analyses identified differences in forward progression between chicks incubated in the two extreme light conditions, supporting the study's hypothesis. We discuss the significance of our findings and potential design considerations for future studies of light-accelerated motor development in precocial and nonprecocial animals.

Drift during overground locomotion in newly hatched chicks varies with light exposure during embryogenesis.

Neurological injuries or disease can impair the function of motor circuitry controlling forearm supination, and recovery is often limited. Preclinical animal models are essential tools for developing therapeutic interventions to improve motor function after neurological damage. Here we describe the supination assessment task, an automated measure of quantifying forelimb supination in the rat.

The supination assessment task: An automated method for quantifying forelimb rotational function in rats.

Rodents are the primary animal model of corticospinal injury and repair, yet current behavioral tests do not show the large deficits after injury observed in humans. Forearm supination is critical for hand function and is highly impaired by corticospinal injury in both humans and rats. Current tests of rodent forelimb function do not measure this movement.

An Automated Test of Rat Forelimb Supination Quantifies Motor Function Loss and Recovery After Corticospinal Injury.

4-Aminopyridine (4-AP) is a Food and Drug Administration-approved drug to improve motor function in people with multiple sclerosis. Preliminary results suggest the drug may act on intact neural circuits and not just on demyelinated ones.

Clinically Relevant Levels of 4-Aminopyridine Strengthen Physiological Responses in Intact Motor Circuits in Rats, Especially After Pyramidal Tract Injury.

Anil Sindhurakar

Burke Medical Research Institute

The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats