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Visual associative learning in restrained honey bees with intact antennae.

Dobrin SE, Fahrbach SE - PLoS ONE (2012)

Bottom Line: Here we report, for a simple visual learning task, the first successful performance by restrained honey bees with intact antennae.A negative correlation was found between age of foragers and their performance in the visual PER task.Using the adaptations to the traditional PER task outlined here, future studies can exploit pharmacological and physiological techniques to explore the neural circuit basis of visual learning in the honey bee.

View Article: PubMed Central - PubMed

Affiliation: Neuroscience Program, Wake Forest University Graduate School of Arts and Sciences, Winston-Salem, North Carolina, United States of America. dobrinse@wfu.edu

ABSTRACT
A restrained honey bee can be trained to extend its proboscis in response to the pairing of an odor with a sucrose reward, a form of olfactory associative learning referred to as the proboscis extension response (PER). Although the ability of flying honey bees to respond to visual cues is well-established, associative visual learning in restrained honey bees has been challenging to demonstrate. Those few groups that have documented vision-based PER have reported that removing the antennae prior to training is a prerequisite for learning. Here we report, for a simple visual learning task, the first successful performance by restrained honey bees with intact antennae. Honey bee foragers were trained on a differential visual association task by pairing the presentation of a blue light with a sucrose reward and leaving the presentation of a green light unrewarded. A negative correlation was found between age of foragers and their performance in the visual PER task. Using the adaptations to the traditional PER task outlined here, future studies can exploit pharmacological and physiological techniques to explore the neural circuit basis of visual learning in the honey bee.

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Description of the experimental paradigm.A. Worker honey bees were restrained in plastic drinking straws using a yoke made of insect pins placed on either side on the neck. Honey bees were supported from below using a rolled paper tissue. A small window was cut in the straw to allow full extension of the proboscis. B. Restrained honey bees were placed in front of individual light presentation screens. Each screen could be illuminated with a blue or green led and had a red LED mounted on top to indicate US presentation to the experimenter. C. A series of projection screens allowed simultaneous conditioning of up to ten honey bees. D. Both the rewarded and unrewarded trials used the same timing of CS/US presentation. Following a 3 sec countdown (not depicted), the CS presentation lasted 5 sec during the final 3 sec of which the US was presented. Proboscis extensions (responses) were recorded to the CS before and during the US presentation.
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pone-0037666-g001: Description of the experimental paradigm.A. Worker honey bees were restrained in plastic drinking straws using a yoke made of insect pins placed on either side on the neck. Honey bees were supported from below using a rolled paper tissue. A small window was cut in the straw to allow full extension of the proboscis. B. Restrained honey bees were placed in front of individual light presentation screens. Each screen could be illuminated with a blue or green led and had a red LED mounted on top to indicate US presentation to the experimenter. C. A series of projection screens allowed simultaneous conditioning of up to ten honey bees. D. Both the rewarded and unrewarded trials used the same timing of CS/US presentation. Following a 3 sec countdown (not depicted), the CS presentation lasted 5 sec during the final 3 sec of which the US was presented. Proboscis extensions (responses) were recorded to the CS before and during the US presentation.

Mentions: Focal foragers (either normal age or precocious) were collected for use in PER studies by placing a wire screen (3 mm spacing) temporarily over the entrance of the hive to prevent honey bees from entering. For collections from the SCCs, individual honey bees were captured in glass vials and immediately placed on ice in the field. For collections of foragers from the typical colony, batches of 15 honey bees were captured in individual glass vials (each batch taking 10–30 min to collect) before being brought into the laboratory and placed on ice. Once immobilized, honey bees were restrained in individual plastic straws (76 mm×13 mm) with the antennae intact. A small window was cut in the straw to allow the proboscis to freely extend. Rolled tissue paper supported the honey bee from below and 2 insect pins were placed through the walls of the straw, on either side on the honey bee’s “neck” to prevent escape as previously described (Fig 1A; [19]). Honey bees were fed 50% sucrose (w/v) ad libitum when they regained movement (approximately 5–10 min after removal from ice) and placed in a dark room (29–32°C) overnight. All subsequent steps were conducted under red light illumination invisible to honey bees [20].


Visual associative learning in restrained honey bees with intact antennae.

Dobrin SE, Fahrbach SE - PLoS ONE (2012)

Description of the experimental paradigm.A. Worker honey bees were restrained in plastic drinking straws using a yoke made of insect pins placed on either side on the neck. Honey bees were supported from below using a rolled paper tissue. A small window was cut in the straw to allow full extension of the proboscis. B. Restrained honey bees were placed in front of individual light presentation screens. Each screen could be illuminated with a blue or green led and had a red LED mounted on top to indicate US presentation to the experimenter. C. A series of projection screens allowed simultaneous conditioning of up to ten honey bees. D. Both the rewarded and unrewarded trials used the same timing of CS/US presentation. Following a 3 sec countdown (not depicted), the CS presentation lasted 5 sec during the final 3 sec of which the US was presented. Proboscis extensions (responses) were recorded to the CS before and during the US presentation.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3368934&req=5

pone-0037666-g001: Description of the experimental paradigm.A. Worker honey bees were restrained in plastic drinking straws using a yoke made of insect pins placed on either side on the neck. Honey bees were supported from below using a rolled paper tissue. A small window was cut in the straw to allow full extension of the proboscis. B. Restrained honey bees were placed in front of individual light presentation screens. Each screen could be illuminated with a blue or green led and had a red LED mounted on top to indicate US presentation to the experimenter. C. A series of projection screens allowed simultaneous conditioning of up to ten honey bees. D. Both the rewarded and unrewarded trials used the same timing of CS/US presentation. Following a 3 sec countdown (not depicted), the CS presentation lasted 5 sec during the final 3 sec of which the US was presented. Proboscis extensions (responses) were recorded to the CS before and during the US presentation.
Mentions: Focal foragers (either normal age or precocious) were collected for use in PER studies by placing a wire screen (3 mm spacing) temporarily over the entrance of the hive to prevent honey bees from entering. For collections from the SCCs, individual honey bees were captured in glass vials and immediately placed on ice in the field. For collections of foragers from the typical colony, batches of 15 honey bees were captured in individual glass vials (each batch taking 10–30 min to collect) before being brought into the laboratory and placed on ice. Once immobilized, honey bees were restrained in individual plastic straws (76 mm×13 mm) with the antennae intact. A small window was cut in the straw to allow the proboscis to freely extend. Rolled tissue paper supported the honey bee from below and 2 insect pins were placed through the walls of the straw, on either side on the honey bee’s “neck” to prevent escape as previously described (Fig 1A; [19]). Honey bees were fed 50% sucrose (w/v) ad libitum when they regained movement (approximately 5–10 min after removal from ice) and placed in a dark room (29–32°C) overnight. All subsequent steps were conducted under red light illumination invisible to honey bees [20].

Bottom Line: Here we report, for a simple visual learning task, the first successful performance by restrained honey bees with intact antennae.A negative correlation was found between age of foragers and their performance in the visual PER task.Using the adaptations to the traditional PER task outlined here, future studies can exploit pharmacological and physiological techniques to explore the neural circuit basis of visual learning in the honey bee.

View Article: PubMed Central - PubMed

Affiliation: Neuroscience Program, Wake Forest University Graduate School of Arts and Sciences, Winston-Salem, North Carolina, United States of America. dobrinse@wfu.edu

ABSTRACT
A restrained honey bee can be trained to extend its proboscis in response to the pairing of an odor with a sucrose reward, a form of olfactory associative learning referred to as the proboscis extension response (PER). Although the ability of flying honey bees to respond to visual cues is well-established, associative visual learning in restrained honey bees has been challenging to demonstrate. Those few groups that have documented vision-based PER have reported that removing the antennae prior to training is a prerequisite for learning. Here we report, for a simple visual learning task, the first successful performance by restrained honey bees with intact antennae. Honey bee foragers were trained on a differential visual association task by pairing the presentation of a blue light with a sucrose reward and leaving the presentation of a green light unrewarded. A negative correlation was found between age of foragers and their performance in the visual PER task. Using the adaptations to the traditional PER task outlined here, future studies can exploit pharmacological and physiological techniques to explore the neural circuit basis of visual learning in the honey bee.

Show MeSH
Related in: MedlinePlus