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PPL2ab neurons restore sexual responses in aged Drosophila males through dopamine.

Kuo SY, Wu CL, Hsieh MY, Lin CT, Wen RK, Chen LC, Chen YH, Yu YW, Wang HD, Su YJ, Lin CJ, Yang CY, Guan HY, Wang PY, Lan TH, Fu TF - Nat Commun (2015)

Bottom Line: We find that increasing DA levels in a subset of cells in the PPL2ab neuronal cluster is necessary and sufficient for increased sustained courtship in both young and aged male flies.Our results indicate that preventing the age-related decline in DA levels in PPL2ab neurons alleviates diminished courtship behaviours in male Drosophila.These results may provide the foundation for deciphering the circuitry involved in sexual motivation in the male Drosophila brain.

View Article: PubMed Central - PubMed

Affiliation: Department of Applied Chemistry, National Chi Nan University, 54561 Nantou, Taiwan.

ABSTRACT
Male sexual desire typically declines with ageing. However, our understanding of the neurobiological basis for this phenomenon is limited by our knowledge of the brain circuitry and neuronal pathways controlling male sexual desire. A number of studies across species suggest that dopamine (DA) affects sexual desire. Here we use genetic tools and behavioural assays to identify a novel subset of DA neurons that regulate age-associated male courtship activity in Drosophila. We find that increasing DA levels in a subset of cells in the PPL2ab neuronal cluster is necessary and sufficient for increased sustained courtship in both young and aged male flies. Our results indicate that preventing the age-related decline in DA levels in PPL2ab neurons alleviates diminished courtship behaviours in male Drosophila. These results may provide the foundation for deciphering the circuitry involved in sexual motivation in the male Drosophila brain.

No MeSH data available.


Related in: MedlinePlus

DAergic PPL2ab neurons regulate male courtship sustainment.Representative images showing the expression pattern of the Gal4 driver lines: (a) murashka-1-Gal4, (b) NP3024-Gal4, (c) NP5945-Gal4 and (d) LG121-LexA; driver expression patterns revealed by UAS-mCD8::GFP (or LexAop-mCD8::GFP) in the adult brain (10-day-old) are shown (green in a–d shows the PPL2ab cell bodies indicated by the arrows and POT track indicated by the arrowheads; the inset box shows the expression pattern in a single calyx). The neuropil was immunostained with an anti-DLG antibody (magenta in a–d). The cell bodies of PPL2ab neurons (green in a1-d1) were labelled with four independent drivers; PPM1/2 neurons (green in a2-c2) were labelled by murashka-1-Gal4, NP3024-Gal4 and NP5945-Gal4; and PPM3 neurons (green in b3) were labelled by the NP3024-Gal4; all of these cell were DAergic as indicated by TH-antibody immunostaining (magenta in a1-2, b1-3, c1-2 and d1). Scale bars, 20 μm. (e,f) The results for courtship index and courtship bout length towards 3-day-old intact CS females or decapitated CS females are shown. ANOVAs were used to compare genetically manipulated 10-day-old male flies (murashka-1-Gal4>UAS-TH, NP3024-Gal4>UAS-TH, NP5945-Gal4>UAS-TH, and LG121-LexA>LexAop-TH) with the corresponding driver and effector heterozygous controls. Significant differences in courtship index (e) and courtship bout length (f) were observed. Each column represents the mean of 18 tests. The error bars indicate+s.e.m. The means for columns followed by the same letters were not significantly different according to the P<0.05 threshold by a one-way ANOVA followed by Tukey's test per grouped columns (separated by a dashed line). (g,h) Two types of PPL2ab neurons from murashka-1-Gal4 flip-out clones (green) and neuropil were immunostained with the anti-DLG antibody (magenta). Genotypes in (a–f): (1) +/y;+/+;murashka-1-Gal4/UAS-TH; (2) +/y;+/+;murashka-1-Gal4/+; (3) +/y;+/+;NP3024-Gal4/UAS-TH; (4) +/y;+/+;NP3024-Gal4/+; (5) +/y;+/+;NP5945-Gal4/UAS-TH; (6) +/y;+/+;NP5945-Gal4/+; (7) +/y;+/+;+/UAS-TH; (8) +/y;LexAop-TH/+;murashka-1-Gal4/+; and (9) +/y;LexAop-TH/+;+/+. Genotypes in (g,h): hs-FLP/y; +/+; UAS>rCD2,y+>mCD8::GFP/murashka-1-Gal4.
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f2: DAergic PPL2ab neurons regulate male courtship sustainment.Representative images showing the expression pattern of the Gal4 driver lines: (a) murashka-1-Gal4, (b) NP3024-Gal4, (c) NP5945-Gal4 and (d) LG121-LexA; driver expression patterns revealed by UAS-mCD8::GFP (or LexAop-mCD8::GFP) in the adult brain (10-day-old) are shown (green in a–d shows the PPL2ab cell bodies indicated by the arrows and POT track indicated by the arrowheads; the inset box shows the expression pattern in a single calyx). The neuropil was immunostained with an anti-DLG antibody (magenta in a–d). The cell bodies of PPL2ab neurons (green in a1-d1) were labelled with four independent drivers; PPM1/2 neurons (green in a2-c2) were labelled by murashka-1-Gal4, NP3024-Gal4 and NP5945-Gal4; and PPM3 neurons (green in b3) were labelled by the NP3024-Gal4; all of these cell were DAergic as indicated by TH-antibody immunostaining (magenta in a1-2, b1-3, c1-2 and d1). Scale bars, 20 μm. (e,f) The results for courtship index and courtship bout length towards 3-day-old intact CS females or decapitated CS females are shown. ANOVAs were used to compare genetically manipulated 10-day-old male flies (murashka-1-Gal4>UAS-TH, NP3024-Gal4>UAS-TH, NP5945-Gal4>UAS-TH, and LG121-LexA>LexAop-TH) with the corresponding driver and effector heterozygous controls. Significant differences in courtship index (e) and courtship bout length (f) were observed. Each column represents the mean of 18 tests. The error bars indicate+s.e.m. The means for columns followed by the same letters were not significantly different according to the P<0.05 threshold by a one-way ANOVA followed by Tukey's test per grouped columns (separated by a dashed line). (g,h) Two types of PPL2ab neurons from murashka-1-Gal4 flip-out clones (green) and neuropil were immunostained with the anti-DLG antibody (magenta). Genotypes in (a–f): (1) +/y;+/+;murashka-1-Gal4/UAS-TH; (2) +/y;+/+;murashka-1-Gal4/+; (3) +/y;+/+;NP3024-Gal4/UAS-TH; (4) +/y;+/+;NP3024-Gal4/+; (5) +/y;+/+;NP5945-Gal4/UAS-TH; (6) +/y;+/+;NP5945-Gal4/+; (7) +/y;+/+;+/UAS-TH; (8) +/y;LexAop-TH/+;murashka-1-Gal4/+; and (9) +/y;LexAop-TH/+;+/+. Genotypes in (g,h): hs-FLP/y; +/+; UAS>rCD2,y+>mCD8::GFP/murashka-1-Gal4.

Mentions: To determine the specific DA neurons that regulate Drosophila courtship sustainment, we used Gal4 drivers to manipulate DA activity in restricted groups of DAergic neurons in the fly brain (Supplementary Fig. 1; murashka-1-Gal4 driver in Fig. 2a; and summarized in Supplementary Table 1). We then analysed the effect of cell-type-specific DA overexpression on courtship intensity in male flies (Fig. 1). Our results revealed that increased expression of TH in TH-Gal4-(ref. 17) and TH-C1-Gal4-positive18 neurons significantly enhanced the male courtship index (Fig. 1). Interestingly, we observed similar results by overexpressing TH in murashka-1-Gal4-positive neurons, which targets a more restricted cell population (Fig. 1). To label the cells targeted by TH-Gal4, TH-C1-Gal4 and murashka-1-Gal4, we crossed each line to the UAS-mCD8::GFP reporter line and counterstained each by TH immunohistochemistry. By assessing the anatomical profile of TH cells coexpressing GFP in each line, we found that expression overlapped between the three Gal4 drivers in a subset of DA neurons in the PPL2ab (Fig. 2a1 and Supplementary Fig. 1g1) and the protocerebral posteriomedial dopaminergic cluster neuron 1/2 (PPM1/2) (Fig. 2a2 and Supplementary Fig. 1g3). These data suggested that male courtship intensity might be regulated by DA release from these neurons.


PPL2ab neurons restore sexual responses in aged Drosophila males through dopamine.

Kuo SY, Wu CL, Hsieh MY, Lin CT, Wen RK, Chen LC, Chen YH, Yu YW, Wang HD, Su YJ, Lin CJ, Yang CY, Guan HY, Wang PY, Lan TH, Fu TF - Nat Commun (2015)

DAergic PPL2ab neurons regulate male courtship sustainment.Representative images showing the expression pattern of the Gal4 driver lines: (a) murashka-1-Gal4, (b) NP3024-Gal4, (c) NP5945-Gal4 and (d) LG121-LexA; driver expression patterns revealed by UAS-mCD8::GFP (or LexAop-mCD8::GFP) in the adult brain (10-day-old) are shown (green in a–d shows the PPL2ab cell bodies indicated by the arrows and POT track indicated by the arrowheads; the inset box shows the expression pattern in a single calyx). The neuropil was immunostained with an anti-DLG antibody (magenta in a–d). The cell bodies of PPL2ab neurons (green in a1-d1) were labelled with four independent drivers; PPM1/2 neurons (green in a2-c2) were labelled by murashka-1-Gal4, NP3024-Gal4 and NP5945-Gal4; and PPM3 neurons (green in b3) were labelled by the NP3024-Gal4; all of these cell were DAergic as indicated by TH-antibody immunostaining (magenta in a1-2, b1-3, c1-2 and d1). Scale bars, 20 μm. (e,f) The results for courtship index and courtship bout length towards 3-day-old intact CS females or decapitated CS females are shown. ANOVAs were used to compare genetically manipulated 10-day-old male flies (murashka-1-Gal4>UAS-TH, NP3024-Gal4>UAS-TH, NP5945-Gal4>UAS-TH, and LG121-LexA>LexAop-TH) with the corresponding driver and effector heterozygous controls. Significant differences in courtship index (e) and courtship bout length (f) were observed. Each column represents the mean of 18 tests. The error bars indicate+s.e.m. The means for columns followed by the same letters were not significantly different according to the P<0.05 threshold by a one-way ANOVA followed by Tukey's test per grouped columns (separated by a dashed line). (g,h) Two types of PPL2ab neurons from murashka-1-Gal4 flip-out clones (green) and neuropil were immunostained with the anti-DLG antibody (magenta). Genotypes in (a–f): (1) +/y;+/+;murashka-1-Gal4/UAS-TH; (2) +/y;+/+;murashka-1-Gal4/+; (3) +/y;+/+;NP3024-Gal4/UAS-TH; (4) +/y;+/+;NP3024-Gal4/+; (5) +/y;+/+;NP5945-Gal4/UAS-TH; (6) +/y;+/+;NP5945-Gal4/+; (7) +/y;+/+;+/UAS-TH; (8) +/y;LexAop-TH/+;murashka-1-Gal4/+; and (9) +/y;LexAop-TH/+;+/+. Genotypes in (g,h): hs-FLP/y; +/+; UAS>rCD2,y+>mCD8::GFP/murashka-1-Gal4.
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f2: DAergic PPL2ab neurons regulate male courtship sustainment.Representative images showing the expression pattern of the Gal4 driver lines: (a) murashka-1-Gal4, (b) NP3024-Gal4, (c) NP5945-Gal4 and (d) LG121-LexA; driver expression patterns revealed by UAS-mCD8::GFP (or LexAop-mCD8::GFP) in the adult brain (10-day-old) are shown (green in a–d shows the PPL2ab cell bodies indicated by the arrows and POT track indicated by the arrowheads; the inset box shows the expression pattern in a single calyx). The neuropil was immunostained with an anti-DLG antibody (magenta in a–d). The cell bodies of PPL2ab neurons (green in a1-d1) were labelled with four independent drivers; PPM1/2 neurons (green in a2-c2) were labelled by murashka-1-Gal4, NP3024-Gal4 and NP5945-Gal4; and PPM3 neurons (green in b3) were labelled by the NP3024-Gal4; all of these cell were DAergic as indicated by TH-antibody immunostaining (magenta in a1-2, b1-3, c1-2 and d1). Scale bars, 20 μm. (e,f) The results for courtship index and courtship bout length towards 3-day-old intact CS females or decapitated CS females are shown. ANOVAs were used to compare genetically manipulated 10-day-old male flies (murashka-1-Gal4>UAS-TH, NP3024-Gal4>UAS-TH, NP5945-Gal4>UAS-TH, and LG121-LexA>LexAop-TH) with the corresponding driver and effector heterozygous controls. Significant differences in courtship index (e) and courtship bout length (f) were observed. Each column represents the mean of 18 tests. The error bars indicate+s.e.m. The means for columns followed by the same letters were not significantly different according to the P<0.05 threshold by a one-way ANOVA followed by Tukey's test per grouped columns (separated by a dashed line). (g,h) Two types of PPL2ab neurons from murashka-1-Gal4 flip-out clones (green) and neuropil were immunostained with the anti-DLG antibody (magenta). Genotypes in (a–f): (1) +/y;+/+;murashka-1-Gal4/UAS-TH; (2) +/y;+/+;murashka-1-Gal4/+; (3) +/y;+/+;NP3024-Gal4/UAS-TH; (4) +/y;+/+;NP3024-Gal4/+; (5) +/y;+/+;NP5945-Gal4/UAS-TH; (6) +/y;+/+;NP5945-Gal4/+; (7) +/y;+/+;+/UAS-TH; (8) +/y;LexAop-TH/+;murashka-1-Gal4/+; and (9) +/y;LexAop-TH/+;+/+. Genotypes in (g,h): hs-FLP/y; +/+; UAS>rCD2,y+>mCD8::GFP/murashka-1-Gal4.
Mentions: To determine the specific DA neurons that regulate Drosophila courtship sustainment, we used Gal4 drivers to manipulate DA activity in restricted groups of DAergic neurons in the fly brain (Supplementary Fig. 1; murashka-1-Gal4 driver in Fig. 2a; and summarized in Supplementary Table 1). We then analysed the effect of cell-type-specific DA overexpression on courtship intensity in male flies (Fig. 1). Our results revealed that increased expression of TH in TH-Gal4-(ref. 17) and TH-C1-Gal4-positive18 neurons significantly enhanced the male courtship index (Fig. 1). Interestingly, we observed similar results by overexpressing TH in murashka-1-Gal4-positive neurons, which targets a more restricted cell population (Fig. 1). To label the cells targeted by TH-Gal4, TH-C1-Gal4 and murashka-1-Gal4, we crossed each line to the UAS-mCD8::GFP reporter line and counterstained each by TH immunohistochemistry. By assessing the anatomical profile of TH cells coexpressing GFP in each line, we found that expression overlapped between the three Gal4 drivers in a subset of DA neurons in the PPL2ab (Fig. 2a1 and Supplementary Fig. 1g1) and the protocerebral posteriomedial dopaminergic cluster neuron 1/2 (PPM1/2) (Fig. 2a2 and Supplementary Fig. 1g3). These data suggested that male courtship intensity might be regulated by DA release from these neurons.

Bottom Line: We find that increasing DA levels in a subset of cells in the PPL2ab neuronal cluster is necessary and sufficient for increased sustained courtship in both young and aged male flies.Our results indicate that preventing the age-related decline in DA levels in PPL2ab neurons alleviates diminished courtship behaviours in male Drosophila.These results may provide the foundation for deciphering the circuitry involved in sexual motivation in the male Drosophila brain.

View Article: PubMed Central - PubMed

Affiliation: Department of Applied Chemistry, National Chi Nan University, 54561 Nantou, Taiwan.

ABSTRACT
Male sexual desire typically declines with ageing. However, our understanding of the neurobiological basis for this phenomenon is limited by our knowledge of the brain circuitry and neuronal pathways controlling male sexual desire. A number of studies across species suggest that dopamine (DA) affects sexual desire. Here we use genetic tools and behavioural assays to identify a novel subset of DA neurons that regulate age-associated male courtship activity in Drosophila. We find that increasing DA levels in a subset of cells in the PPL2ab neuronal cluster is necessary and sufficient for increased sustained courtship in both young and aged male flies. Our results indicate that preventing the age-related decline in DA levels in PPL2ab neurons alleviates diminished courtship behaviours in male Drosophila. These results may provide the foundation for deciphering the circuitry involved in sexual motivation in the male Drosophila brain.

No MeSH data available.


Related in: MedlinePlus