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Schaeffer, L.

Publications and source records attributed to Schaeffer, L..

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Methods for chair restraint and training of the common marmoset on oculomotor tasks

The oculomotor system is the most thoroughly understood sensorimotor system in the brain, due in large part to electrophysiological studies carried out in macaque monkeys trained to perform ocuolomotor tasks. A disadvantage of the macaque model is that many cortical oculomotor areas of interest lie within sulci, making high-density array and laminar recordings impractical. Further, many techniques of molecular biology developed in rodents, such as transgenic animals and optogenetic manipulation of neuronal subtypes, are limited in this species. The common marmoset (Callithrix jacchus) may potentially bridge the gap between systems neuroscience in macaques and molecular techniques, and additionally possesses a smooth cortex allowing easy access to frontoparietal oculomotor areas. To date, techniques for restraint and training of these animals to perform oculomotor tasks remain in an early stage of development. Here we provide details of a custom-designed chair for restraint of marmosets, a combination head restraint/recording chamber providing stability suitable for eye movement and neural recordings, and a training protocol for oculomotor tasks. As proof-of-principle, we report the results of a psychophysical study in marmosets trained to perform a saccade task using these methods, showing that, as in rhesus and humans, marmosets exhibit a \"gap effect\" - a decrease in reaction time when the fixation stimulus is removed prior to the onset of a visual saccade target. These results provide evidence that the common marmoset is a suitable model for neurophysiogical investigations of oculomotor control.\n\nNEW AND NOTEWORTHYThe ability to carry out neuronal recordings in behaving primates has provided a wealth of information regarding the neural circuits underlying the control of eye movements. Such studies require restraint of the animal within a primate chair, head fixation, methods of acclimating the animals to this restraint, and the use of operant conditioning methods for training on oculomotor tasks. In contrast to the macaque model, relatively few studies have reported in detail methods for use in the common marmoset. Here we detail custom-designed equipment and methods by which we have used to successfully train head-restrained marmosets to perform basic oculomotor tasks.

neuroscience

Efficient genome editing in primary cells and in vivo using viral-derived "Nanoblades" loaded with Cas9/sgRNA ribonucleoproteins

Programmable nucleases have enabled rapid and accessible genome engineering in eukaryotic cells and living organisms. However, their delivery into target cells can be technically challenging when working with primary cells or in vivo. Using engineered murine leukemia virus-like particles loaded with Cas9/sgRNA ribonucleoproteins (\"Nanoblades\"), we were able to induce efficient genome-editing in cell lines and primary cells including human induced pluripotent stem cells, human hematopoietic stem cells and mouse bone-marrow cells. Transgene-free Nanoblades were also capable of in vivo genome-editing in mouse embryos and in the liver of injected mice. Nanoblades can be complexed with donor DNA for \"all-in-one\" homology-directed repair or programmed with modified Cas9 variants to mediate transcriptional up-regulation of target genes. Nanoblades preparation process is simple, relatively inexpensive and can be easily implemented in any laboratory equipped for cellular biology.

cell biology