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Effects of Flight on Gene Expression and Aging in the Honey Bee Brain and Flight Muscle.

Margotta JW, Mancinelli GE, Benito AA, Ammons A, Roberts SP, Elekonich MM - Insects (2012)

Bottom Line: To investigate the effects of flight, behavioral state and age on gene expression, we used whole-genome microarrays and real-time PCR.Our data suggest that the transition from behaviors requiring little to no flight (nursing) to those requiring prolonged flight bouts (foraging), rather than the amount of previous flight per se, has a major effect on gene expression.Combined with our real-time PCR data, these data suggest an epigenetic control and energy balance role in honey bee functional senescence.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, University of Nevada, Las Vegas, NV 89154, USA. margotta@unlv.nevada.edu.

ABSTRACT
Honey bees move through a series of in-hive tasks (e.g., "nursing") to outside tasks (e.g., "foraging") that are coincident with physiological changes and higher levels of metabolic activity. Social context can cause worker bees to speed up or slow down this process, and foragers may revert back to their earlier in-hive tasks accompanied by reversion to earlier physiological states. To investigate the effects of flight, behavioral state and age on gene expression, we used whole-genome microarrays and real-time PCR. Brain tissue and flight muscle exhibited different patterns of expression during behavioral transitions, with expression patterns in the brain reflecting both age and behavior, and expression patterns in flight muscle being primarily determined by age. Our data suggest that the transition from behaviors requiring little to no flight (nursing) to those requiring prolonged flight bouts (foraging), rather than the amount of previous flight per se, has a major effect on gene expression. Following behavioral reversion there was a partial reversion in gene expression but some aspects of forager expression patterns, such as those for genes involved in immune function, remained. Combined with our real-time PCR data, these data suggest an epigenetic control and energy balance role in honey bee functional senescence.

No MeSH data available.


Microarray experimental design. The microarray experiment was designed with 132 arrays in a “round-robin” style. Each arrow represents 6 arrays. YN = young nurse (10 days-old; <1 day flight); RN = reverted nurse (25–26 days-old; 7–9 days flight); OF = old forager (25–26 days-old; 10–12 days flight); TH = typical-aged forager-high flight (19–22 days old; 7–9 days flight); ON = older nurse (19–22 days old; <1 day flight); TL = typical-aged forager-low flight (19–22 days old; 2–3 days flight); PF = precocious forager (8–10 days old; 2–3 days flight).
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insects-04-00009-f001: Microarray experimental design. The microarray experiment was designed with 132 arrays in a “round-robin” style. Each arrow represents 6 arrays. YN = young nurse (10 days-old; <1 day flight); RN = reverted nurse (25–26 days-old; 7–9 days flight); OF = old forager (25–26 days-old; 10–12 days flight); TH = typical-aged forager-high flight (19–22 days old; 7–9 days flight); ON = older nurse (19–22 days old; <1 day flight); TL = typical-aged forager-low flight (19–22 days old; 2–3 days flight); PF = precocious forager (8–10 days old; 2–3 days flight).

Mentions: Brains and thoraces from the same individuals of all behavioral groups were compared on a total of 132 arrays. The samples were hybridized against each other using a loop design [36] (See Figure 1 for experiment design). The microarray hybridization procedure followed previously described methods [37]. Extracted total RNA, cDNA, and labeled aRNA sample concentrations were quantified using a Nanodrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). Five hundred nanograms of RNA were amplified using the MessageAmp II aRNA amplification kit (Ambion, Austin, TX, USA). For each individual, 4 micrograms of brain RNA were labeled with cy3 and 4 micrograms of thorax RNA were labeled with cy5 using a Kreatech Labeling Kit (Applied Biosystems, Salt Lake City, UT, USA). The directionality of the dye labeling was switched between replicates to avoid a dye bias. Whole genome oligonucleotide arrays (W.M Keck Center for Comparative and Functional Genomics, University of Illinois, Urbana-Champaign) were hybridized with 120 picomoles of each labeled tissue’s probes. Arrays were then scanned with a GenePix scanner (Molecular Devices, Sunnyvale, CA, USA) and visualized using GenePix software (Agilent Technologies, Santa Clara, CA, USA). Prior to data analysis, each array was visually inspected for any inconsistencies in dye incorporation.


Effects of Flight on Gene Expression and Aging in the Honey Bee Brain and Flight Muscle.

Margotta JW, Mancinelli GE, Benito AA, Ammons A, Roberts SP, Elekonich MM - Insects (2012)

Microarray experimental design. The microarray experiment was designed with 132 arrays in a “round-robin” style. Each arrow represents 6 arrays. YN = young nurse (10 days-old; <1 day flight); RN = reverted nurse (25–26 days-old; 7–9 days flight); OF = old forager (25–26 days-old; 10–12 days flight); TH = typical-aged forager-high flight (19–22 days old; 7–9 days flight); ON = older nurse (19–22 days old; <1 day flight); TL = typical-aged forager-low flight (19–22 days old; 2–3 days flight); PF = precocious forager (8–10 days old; 2–3 days flight).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4553427&req=5

insects-04-00009-f001: Microarray experimental design. The microarray experiment was designed with 132 arrays in a “round-robin” style. Each arrow represents 6 arrays. YN = young nurse (10 days-old; <1 day flight); RN = reverted nurse (25–26 days-old; 7–9 days flight); OF = old forager (25–26 days-old; 10–12 days flight); TH = typical-aged forager-high flight (19–22 days old; 7–9 days flight); ON = older nurse (19–22 days old; <1 day flight); TL = typical-aged forager-low flight (19–22 days old; 2–3 days flight); PF = precocious forager (8–10 days old; 2–3 days flight).
Mentions: Brains and thoraces from the same individuals of all behavioral groups were compared on a total of 132 arrays. The samples were hybridized against each other using a loop design [36] (See Figure 1 for experiment design). The microarray hybridization procedure followed previously described methods [37]. Extracted total RNA, cDNA, and labeled aRNA sample concentrations were quantified using a Nanodrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). Five hundred nanograms of RNA were amplified using the MessageAmp II aRNA amplification kit (Ambion, Austin, TX, USA). For each individual, 4 micrograms of brain RNA were labeled with cy3 and 4 micrograms of thorax RNA were labeled with cy5 using a Kreatech Labeling Kit (Applied Biosystems, Salt Lake City, UT, USA). The directionality of the dye labeling was switched between replicates to avoid a dye bias. Whole genome oligonucleotide arrays (W.M Keck Center for Comparative and Functional Genomics, University of Illinois, Urbana-Champaign) were hybridized with 120 picomoles of each labeled tissue’s probes. Arrays were then scanned with a GenePix scanner (Molecular Devices, Sunnyvale, CA, USA) and visualized using GenePix software (Agilent Technologies, Santa Clara, CA, USA). Prior to data analysis, each array was visually inspected for any inconsistencies in dye incorporation.

Bottom Line: To investigate the effects of flight, behavioral state and age on gene expression, we used whole-genome microarrays and real-time PCR.Our data suggest that the transition from behaviors requiring little to no flight (nursing) to those requiring prolonged flight bouts (foraging), rather than the amount of previous flight per se, has a major effect on gene expression.Combined with our real-time PCR data, these data suggest an epigenetic control and energy balance role in honey bee functional senescence.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, University of Nevada, Las Vegas, NV 89154, USA. margotta@unlv.nevada.edu.

ABSTRACT
Honey bees move through a series of in-hive tasks (e.g., "nursing") to outside tasks (e.g., "foraging") that are coincident with physiological changes and higher levels of metabolic activity. Social context can cause worker bees to speed up or slow down this process, and foragers may revert back to their earlier in-hive tasks accompanied by reversion to earlier physiological states. To investigate the effects of flight, behavioral state and age on gene expression, we used whole-genome microarrays and real-time PCR. Brain tissue and flight muscle exhibited different patterns of expression during behavioral transitions, with expression patterns in the brain reflecting both age and behavior, and expression patterns in flight muscle being primarily determined by age. Our data suggest that the transition from behaviors requiring little to no flight (nursing) to those requiring prolonged flight bouts (foraging), rather than the amount of previous flight per se, has a major effect on gene expression. Following behavioral reversion there was a partial reversion in gene expression but some aspects of forager expression patterns, such as those for genes involved in immune function, remained. Combined with our real-time PCR data, these data suggest an epigenetic control and energy balance role in honey bee functional senescence.

No MeSH data available.