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First published online March 14, 2005
Journal of Experimental Biology 208, 1147-1159 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01508

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Three-dimensional antennal lobe atlas of male and female moths, Lobesia botrana (Lepidoptera: Tortricidae) and glomerular representation of plant volatiles in females

Ingwild Masante-Roca¹, Christophe Gadenne¹ and Sylvia Anton^1,2,3,*

¹ Institut National de la Recherche Agronomique, Unité Mixte de Recherche en Santé Végétale, Centre de Recherche de Bordeaux, BP81, 33883 Villenave d'Ornon Cedex, France
² Institut National de la Recherche Agronomique, Unité de Phytopharmacie et de Médiateurs Chimiques, Centre de Recherche de Versailles, 78026 Versailles Cedex, France
³ Department of Chemical Ecology, Swedish Agricultural University, S-230 53 Alnarp, Sweden

View larger version (77K): [in a new window]   Fig. 6. Physiological and anatomical characteristics of broadly responding PNs in Lobesia botrana. (A) Example for a PN with excitatory responses observed for all tested compounds at 1 µg. Note the different delays in response to different compounds. This neuron is not included in the data set in Table 1, because the AL was damaged and the target glomerulus could not be identified. (B) Extended focus image of a stack of confocal sections through the intracellularly stained dendritic arborisations of the generalist PN 14 in glomerulus 37 (black outline). (C) Partial three-dimensional reconstruction of the AL containing the stained PN 14 in the anterior view Y+, indicating the innervated glomerulus 37 (black outline). Brain sketch indicates the orientation of the brain and the relative position of the AL (marked red). AN antennal nerve, DB dorsal border of the AL, OL optic lobe, SOG suboesophageal ganglion, numbers indicate ordinary glomeruli, bold numbers indicate landmark glomeruli. Horizontal scale bar in A, 500 ms, vertical scale bar in A, 40 mV. Scale bar in B 30 µm. Scale bar in C 30 µm.

View larger version (83K): [in a new window]   Fig. 1. Series of frontal confocal sections through the antennal lobe of a Lobesia botrana female (A) and male (B) from anterior (left) to posterior (right) (see also reconstruction in Figs 2, 3, 4, 5), AN antennal nerve, CB cell body cluster, d dorsal, l lateral, m medial, MG macroglomerulus, v, ventral, numbers indicate ordinary glomeruli, bold numbers indicate landmark glomeruli. Note that especially posterior glomeruli are difficult to identify. Scale bar in A and B 30 µm.

View larger version (46K): [in a new window]   Fig. 2. Three-dimensional reconstruction of the left antennal lobe of a female Lobesia botrana (same brain as in Fig. 1A). Graphs in the first row show the projection onto the frontal orientation plane Y. Graphs in the second row show levels S1 and S2 in the Y+ orientation plane. Graphs in the third row show the projection onto the horizontal orientation plane Z. Graphs in the fourth row show the projection onto the sagittal orientation plane X. Brain sketches show the orientation of the brain and the relative position of the AL (marked red) for each view. In the lateral view, the AN reconstruction was omitted and in the dorsal view the reconstruction of the dorsal border of the AL was omitted to visualize the glomeruli hidden otherwise. AN antennal nerve, DB dorsal border of the AL, OL optic lobe, SOG suboesophageal ganglion, numbers indicate ordinary glomeruli, bold numbers indicate landmark glomeruli. Scale bar 20 µm.

View larger version (38K): [in a new window]   Fig. 3. Three-dimensional reconstruction of the right antennal lobes in two female Lobesia botrana individuals. Each vertical column marked with frames of the same colour corresponds to one brain, AL on the left is from the same brain as AL in figure 2. Graphs show the projection onto the frontal orientation plane Y+ viewed from the anterior side. Different sections through the reconstruction of the left lobe are shown to visualize the differently coloured layers of glomeruli. The anterior view shows mainly pink and yellow glomeruli, section S1 shows orange and some red glomeruli, section S2 the remaining red and most green glomeruli and section S3 shows the remaining green and blue glomeruli. Brain sketches indicate the orientation of the brain, the relative position of the AL (marked red) and the level of the sections (dotted lines). AN antennal nerve, DB dorsal border of the AL, OL optic lobe, SOG suboesophageal ganglion, numbers indicate ordinary glomeruli, bold numbers indicate landmark glomeruli. Scale bar 20 µm.

View larger version (47K): [in a new window]   Fig. 4. Three-dimensional reconstruction of the left antennal lobe of a male Lobesia botrana in six different views (same brain as in Fig. 1B). Graphs in the first row show the projection onto the frontal orientation plane Y. Graphs in the second row show the projection onto the horizontal orientation plane Z. Graphs in the third row show the projection onto the sagittal orientation plane X. Brain sketches show the orientation of the brain and the relative position of the AL (marked red) for each view. In the lateral view, the AN reconstruction was omitted and in the dorsal view the reconstruction of the dorsal border of the AL was omitted to visualize the glomeruli hidden otherwise. AN antennal nerve, DB dorsal border of the AL, MG macroglomerulus, OL optic lobe, SOG suboesophageal ganglion, numbers indicate ordinary glomeruli, bold numbers indicate landmark glomeruli. Scale bar 30 µm.

View larger version (38K): [in a new window]   Fig. 5. Three-dimensional reconstruction of the left and right antennal lobe of a male Lobesia botrana (same brain as in Figs 1B and 4). Graphs show the projection onto the frontal orientation plane Y viewed from the anterior side. Different sections through the reconstruction of the left lobe are shown to visualize the differently coloured layers of glomeruli. The anterior view shows mainly pink and yellow glomeruli, section S1 shows orange and some red glomeruli, section S2 the remaining red and most green glomeruli and section S3 shows the remaining green and blue glomeruli. Brain sketches indicate the orientation of the brain, the relative position of the AL (marked red) and the level of the sections (dotted lines). AN antennal nerve, DB dorsal border of the AL, MG macroglomerulus, OL optic lobe, SOG suboesophageal ganglion, numbers indicate ordinary glomeruli, bold numbers indicate landmark glomeruli. Scale bar 30 µm.

View larger version (31K): [in a new window]   Fig. 7. Partial three-dimensional reconstructions of the ALs containing stained PNs in Lobesia botrana in the anterior view Y+, indicating the innervated glomeruli (black outline). (A) Two PNs responding exclusively to -farnesene arborised in two clearly different glomeruli (9 and 5). (B) In two preparations with PNs responding to nonatriene at a lower threshold (PN4) or exclusively to nonatriene (PN12), one PN (PN4) arborised in glomerulus 1, and in the other preparation (PNs 12 a, b) a double staining was found (8, 19). (C) Two PNs arborising in the easily identifiable glomerulus 3, responded to -farnesene and ß-thujone respectively. Brain sketches indicate the orientation of the brain and the relative position of the AL (marked red). AN antennal nerve, DB dorsal border of the AL, OL optic lobe, SOG suboesophageal ganglion, numbers indicate ordinary glomeruli, bold numbers indicate landmark glomeruli. Scale bar in each reconstruction 20 µm.

View larger version (40K): [in a new window]   Fig. 8. Three-dimensional map of the female Lobesia botrana antennal lobe (see Fig. 2) indicating dendritic arborisations of physiologically characterised PNs found in the present study. All target glomeruli of PNs responding to a given odour are marked in grey with white numbers. Only the target glomerulus of the generalist PN is not indicated, as it is situated in a lower (red) level, whereas all other glomeruli were found close to the anterior surface of the AL. AN antennal nerve, DB dorsal border of the AL, OL optic lobe, SOG suboesophageal ganglion, numbers indicate ordinary glomeruli, bold numbers indicate landmark glomeruli. Scale bar 20 µm.

View larger version (77K): [in a new window]   Fig. 9. Anatomy of a local interneuron, which responded only to -farnesene. (A) The reconstruction of the entire neuron in an approximately frontal view shows that arborisations are much more dense in one glomerulus (36) than in the remaining glomeruli (see also B). (B) Single approximately frontal confocal section showing dense arborisations in glomerulus 36. (C) Single approximately frontal confocal section through the central part of the AL showing sparse, branching with varicosities in many glomeruli. AN antennal nerve, numbers indicate ordinary glomeruli, bold numbers indicate landmark glomeruli. Scale bars 30 µm.