Barbara Heider

Two-Photon Imaging of Calcium in Virally Transfected Striate Cortical Neurons of Behaving Monkey

Two-photon scanning microscopy has advanced our understanding of neural signaling in non-mammalian species and mammals. Various developments are needed to perform two-photon scanning microscopy over prolonged periods in non-human primates performing a behavioral task. In striate cortex in two macaque monkeys, cortical neurons were transfected with a genetically encoded fluorescent calcium sensor, memTNXL, using AAV1 as a viral vector. By constructing an extremely rigid and stable apparatus holding both the two-photon scanning microscope and the monkeys head, single neurons were imaged at high magnification for prolonged periods with minimal motion artifacts for up to ten months. Structural images of single neurons were obtained at high magnification. Changes in calcium during visual stimulation were measured as the monkeys performed a fixation task. Overall, functional responses and orientation tuning curves were obtained in 18.8% of the 234 labeled and imaged neurons. This demonstrated that the two-photon scanning microscopy can be successfully obtained in behaving primates.

Two-photon scanning microscopy of in vivo sensory responses of cortical neurons genetically encoded with a fluorescent voltage sensor in rat

A fluorescent voltage sensor protein “Flare” was created from a Kv1.4 potassium channel with YFP situated to report voltage-induced conformational changes in vivo. The RNA virus Sindbis introduced Flare into neurons in the binocular region of visual cortex in rat. Injection sites were selected based on intrinsic optical imaging. Expression of Flare occurred in the cell bodies and dendritic processes. Neurons imaged in vivo using two-photon scanning microscopy typically revealed the soma best, discernable against the background labeling of the neuropil. Somatic fluorescence changes were correlated with flashed visual stimuli; however, averaging was essential to observe these changes. This study demonstrates that the genetic modification of single neurons to express a fluorescent voltage sensor can be used to assess neuronal activity in vivo.

Neural Representation During Visually Guided Reaching in Macaque Posterior Parietal Cortex

Visually guided hand movements in primates require an interconnected network of various cortical areas. Single unit firing rate from area 7a and dorsal prelunate (DP) neurons of macaque posterior parietal cortex (PPC) was recorded during reaching movements to targets at variable locations and under different eye position conditions. In the eye position-varied task, the reach target was always foveated; thus eye position varied with reach target location. In the retinal-varied task, the monkey reached to targets at variable retinotopic locations while eye position was kept constant in the center. Spatial tuning was examined with respect to temporal (task epoch) and contextual (task condition) aspects, and response fields were compared. The analysis showed distinct tuning types. The majority of neurons changed their gain field tuning and retinotopic tuning between different phases of the task. Between the onset of visual stimulation and the preparatory phase (before the go signal), about one half the neurons altered their firing rate significantly. Spatial response fields during preparation and initiation epochs were strongly influenced by the task condition (eye position varied vs. retinal varied), supporting a strong role of eye position during visually guided reaching. DP neurons, classically considered visual, showed reach related modulation similar to 7a neurons. This study shows that both area 7a and DP are modulated during reaching behavior in primates. The various tuning types in both areas suggest distinct populations recruiting different circuits during visually guided reaching.

Spatial effects of shifting prisms on properties of posterior parietal cortex neurons

The posterior parietal cortex contains multiple spatial representations and is involved in online monitoring of visually guided hand movements. Single unit recordings were performed in two areas of macaque monkey posterior parietal cortex during a visually guided reaching task with variable eye position. To test the adaptability of neural responses, shifting prisms were introduced to create a discrepancy between perceived and actual reach location. The majority of neurons changed average firing rate and/or eye position tuning during the prism exposure. The direction of tuning change did not correlate with the direction of prism shift, suggesting more generalized network effects due to the perturbation. Population analysis using Euler angles and translations demonstrated systematic transformations between conditions supporting the notion of network behaviour.

Optical imaging of visually guided reaching in macaque posterior parietal cortex

Sensorimotor transformation for reaching movements in primates requires a large network of visual, parietal, and frontal cortical areas. We performed intrinsic optical imaging over posterior parietal cortex including areas 7a and the dorsal perilunate in macaque monkeys during visually guided hand movements. Reaching was performed while foveating one of nine static reach targets; thus eye-position-varied concurrently with reach position. The hemodynamic reflectance signal was analyzed during specific phases of the task including pre-reach, reach, and touch epochs. The eye position maps changed substantially as the task progressed: First, direction of spatial tuning shifted from a weak preference close to the center to the lower eye positions in both cortical areas. Overall tuning strength was greater in area 7a. Second, strength of spatial tuning increased from the early pre-reach to the later touch epoch. These consistent temporal changes suggest that dynamic properties of the reflectance signal were modulated by task parameters. The peak amplitude and peak delay of the reflectance signal showed considerable differences between eye position but were similar between areas. Compared with a detection task using a lever response, the reach task yielded higher amplitudes and longer delays. These findings demonstrate a spatially tuned topographical representation for reaching in both areas and suggest a strong synergistic combination of various feedback signals that result in a spatially tuned amplification of the hemodynamic response in posterior parietal cortex.