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Viral vector-mediated selective and reversible blockade of the pathway for visual orienting in mice.

Sooksawate T, Isa K, Matsui R, Kato S, Kinoshita M, Kobayashi K, Watanabe D, Kobayashi K, Isa T - Front Neural Circuits (2013)

Bottom Line: A neuron-specific retrograde gene transfer vector with the gene encoding enhanced tetanus neurotoxin (eTeNT) tagged with enhanced green fluorescent protein (EGFP) under the control of a tetracycline responsive element was injected into the left medial pontine reticular formation. 7-17 days later, an adeno-associated viral vector with a highly efficient Tet-ON sequence, rtTAV16, was injected into the right superior colliculus. 5-9 weeks later, the daily administration of doxycycline (Dox) was initiated.Anti-GFP immunohistochemistry revealed that a number of neurons in the intermediate and deep layers of the right superior colliculus were positively stained, indicating eTeNT expression.A second round of Dox administration, starting from 28 days after the termination of the first Dox administration, resulted in the reappearance of the behavioral impairment.

View Article: PubMed Central - PubMed

Affiliation: Department of Developmental Physiology, National Institute for Physiological Sciences Okazaki, Japan ; Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University Bangkok, Thailand.

ABSTRACT
Recently, by using a combination of two viral vectors, we developed a technique for pathway-selective and reversible synaptic transmission blockade, and successfully induced a behavioral deficit of dexterous hand movements in macaque monkeys by affecting a population of spinal interneurons. To explore the capacity of this technique to work in other pathways and species, and to obtain fundamental methodological information, we tried to block the crossed tecto-reticular pathway, which is known to control orienting responses to visual targets, in mice. A neuron-specific retrograde gene transfer vector with the gene encoding enhanced tetanus neurotoxin (eTeNT) tagged with enhanced green fluorescent protein (EGFP) under the control of a tetracycline responsive element was injected into the left medial pontine reticular formation. 7-17 days later, an adeno-associated viral vector with a highly efficient Tet-ON sequence, rtTAV16, was injected into the right superior colliculus. 5-9 weeks later, the daily administration of doxycycline (Dox) was initiated. Visual orienting responses toward the left side were impaired 1-4 days after Dox administration. Anti-GFP immunohistochemistry revealed that a number of neurons in the intermediate and deep layers of the right superior colliculus were positively stained, indicating eTeNT expression. After the termination of Dox administration, the anti-GFP staining returned to the baseline level within 28 days. A second round of Dox administration, starting from 28 days after the termination of the first Dox administration, resulted in the reappearance of the behavioral impairment. These findings showed that pathway-selective and reversible blockade of synaptic transmission also causes behavioral effects in rodents, and that the crossed tecto-reticular pathway clearly controls visual orienting behaviors.

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Related in: MedlinePlus

(A) A photograph showing the visual orienting response of a mouse toward the unaffected side. (B) Effect of the Tet-induced tetanus neurotoxin-expressing system on the visual orienting response of the affected side after the 1st Dox administration for 7 days, termination of Dox administration for 28 days, and the 2nd Dox administration for 7 days.
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Figure 3: (A) A photograph showing the visual orienting response of a mouse toward the unaffected side. (B) Effect of the Tet-induced tetanus neurotoxin-expressing system on the visual orienting response of the affected side after the 1st Dox administration for 7 days, termination of Dox administration for 28 days, and the 2nd Dox administration for 7 days.

Mentions: A visual orienting response test was used to evaluate visual orienting behavior toward a visual target presented either on the affected or unaffected side of the tested mouse. The mouse was placed inside a small glass cylinder (15 cm height, 14 cm inner diameter), and a stimulus (a small long-handle stainless steel chemical spoon) was quickly moved into the visual field of the mouse from behind, in the same horizontal plane as its eyes, until the stimulus was located between its eyes (Smith et al., 1998). Normally, when the stimulus entered its visual field, the mouse turned its head toward the stimulus (Figure 3A). The procedure was conducted bilaterally in a pseudorandom sequence (10 trials per side each day). The number of times the mouse successfully oriented itself to the stimulus was counted.


Viral vector-mediated selective and reversible blockade of the pathway for visual orienting in mice.

Sooksawate T, Isa K, Matsui R, Kato S, Kinoshita M, Kobayashi K, Watanabe D, Kobayashi K, Isa T - Front Neural Circuits (2013)

(A) A photograph showing the visual orienting response of a mouse toward the unaffected side. (B) Effect of the Tet-induced tetanus neurotoxin-expressing system on the visual orienting response of the affected side after the 1st Dox administration for 7 days, termination of Dox administration for 28 days, and the 2nd Dox administration for 7 days.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3795302&req=5

Figure 3: (A) A photograph showing the visual orienting response of a mouse toward the unaffected side. (B) Effect of the Tet-induced tetanus neurotoxin-expressing system on the visual orienting response of the affected side after the 1st Dox administration for 7 days, termination of Dox administration for 28 days, and the 2nd Dox administration for 7 days.
Mentions: A visual orienting response test was used to evaluate visual orienting behavior toward a visual target presented either on the affected or unaffected side of the tested mouse. The mouse was placed inside a small glass cylinder (15 cm height, 14 cm inner diameter), and a stimulus (a small long-handle stainless steel chemical spoon) was quickly moved into the visual field of the mouse from behind, in the same horizontal plane as its eyes, until the stimulus was located between its eyes (Smith et al., 1998). Normally, when the stimulus entered its visual field, the mouse turned its head toward the stimulus (Figure 3A). The procedure was conducted bilaterally in a pseudorandom sequence (10 trials per side each day). The number of times the mouse successfully oriented itself to the stimulus was counted.

Bottom Line: A neuron-specific retrograde gene transfer vector with the gene encoding enhanced tetanus neurotoxin (eTeNT) tagged with enhanced green fluorescent protein (EGFP) under the control of a tetracycline responsive element was injected into the left medial pontine reticular formation. 7-17 days later, an adeno-associated viral vector with a highly efficient Tet-ON sequence, rtTAV16, was injected into the right superior colliculus. 5-9 weeks later, the daily administration of doxycycline (Dox) was initiated.Anti-GFP immunohistochemistry revealed that a number of neurons in the intermediate and deep layers of the right superior colliculus were positively stained, indicating eTeNT expression.A second round of Dox administration, starting from 28 days after the termination of the first Dox administration, resulted in the reappearance of the behavioral impairment.

View Article: PubMed Central - PubMed

Affiliation: Department of Developmental Physiology, National Institute for Physiological Sciences Okazaki, Japan ; Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University Bangkok, Thailand.

ABSTRACT
Recently, by using a combination of two viral vectors, we developed a technique for pathway-selective and reversible synaptic transmission blockade, and successfully induced a behavioral deficit of dexterous hand movements in macaque monkeys by affecting a population of spinal interneurons. To explore the capacity of this technique to work in other pathways and species, and to obtain fundamental methodological information, we tried to block the crossed tecto-reticular pathway, which is known to control orienting responses to visual targets, in mice. A neuron-specific retrograde gene transfer vector with the gene encoding enhanced tetanus neurotoxin (eTeNT) tagged with enhanced green fluorescent protein (EGFP) under the control of a tetracycline responsive element was injected into the left medial pontine reticular formation. 7-17 days later, an adeno-associated viral vector with a highly efficient Tet-ON sequence, rtTAV16, was injected into the right superior colliculus. 5-9 weeks later, the daily administration of doxycycline (Dox) was initiated. Visual orienting responses toward the left side were impaired 1-4 days after Dox administration. Anti-GFP immunohistochemistry revealed that a number of neurons in the intermediate and deep layers of the right superior colliculus were positively stained, indicating eTeNT expression. After the termination of Dox administration, the anti-GFP staining returned to the baseline level within 28 days. A second round of Dox administration, starting from 28 days after the termination of the first Dox administration, resulted in the reappearance of the behavioral impairment. These findings showed that pathway-selective and reversible blockade of synaptic transmission also causes behavioral effects in rodents, and that the crossed tecto-reticular pathway clearly controls visual orienting behaviors.

Show MeSH
Related in: MedlinePlus