Cell and Tissue Research

, Volume 343, Issue 2, pp 357–369 | Cite as

A novel wide-field neuron with branches in the lamina of the Drosophila visual system expresses myoinhibitory peptide and may be associated with the clock

  • Agata Kolodziejczyk
  • Dick R. NässelEmail author
Regular Article


Although neuropeptides are widespread throughout the central nervous system of the fruifly Drosophila, no records exist of peptidergic neurons in the first synaptic region of the visual system, the lamina. Here, we describe a novel type of neuron that has wide-field tangential arborizations just distal to the lamina neuropil and that expresses myoinhibitory peptide (MIP). The cell bodies of these neurons, designated lateral MIP-immunoreactive optic lobe (LMIo) neurons, lie anteriorly at the base of the medulla of the optic lobe. The LMIo neurons also arborize in several layers of the medulla and in the dorso-lateral and lateral protocerebrum. Since the LMIo resemble LNv clock neurons, we have investigated the relationships between these two sets of neurons by combining MIP-immunolabeling with markers for two of the clock genes, viz., Cryptochrome and Timeless, or with antisera to two peptides expressed in clock neurons, viz., pigment-dispersing factor and ion transport peptide. LMIo neurons do not co-express any of these clock neuron markers. However, branches of LMIo and clock neurons overlap in several regions. Furthermore, the varicose lamina branches of LMIo neurons superimpose those of two large bilateral serotonergic neurons. The close apposition of the terminations of MIP- and serotonin-producing neurons distal to the lamina suggests that they have the same peripheral targets. Our data indicate that the LMIo neurons are not bona fide clock neurons, but they may be associated with the clock system and regulate signaling peripherally in the visual system.


Neuropeptide Allatostatin B Serotonin Biological clock Optic lobe Insect brain Drososphila melanogaster (Insecta) 



We thank Dr. Lily Kahsai for her important participation in initial experiments on MIP neurons in the visual system. The individuals and organizations listed in the Materials and methods are gratefully acknowledged for supplying fly strains and reagents.

Supplementary material

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S. Fig. 1

Serotonin-immunoreactive (IR) neurons in relation to MIP-IR neurons (rabbit anti-MIP). a Varicose serotonin-IR processes (magenta) are found in a layer (X) between the cell bodies (Cb) of monopolar neurons and the lamina synaptic layer (La). As a marker for synaptic sites, a mouse monoclonal antibody to Discs large (DLG) was used (green). The serotonergic boutons thus reside outside the region in which photoreceptors and lamina neurons form synapses. b Posterior portion of the brain (frontal view) with the two large cell bodies of the LBO5HT neurons (arrows) and their neurites (arrowheads). Their main bilateral axons (ax) run into the optic lobe, including the lamina region. c1-3 Optic lobe labeled with antisera to MIP (magenta) and serotonin (green). Frontal section (dorsal is to the right, lateral to the top). The three lateral MIP-immunoreactive optic lobe (LMIo) neurons (star) innervate the medulla (Me) and medio-lateral protocerebrum (MLP) and lamina (not shown here). The pattern of serotonin-IR (5-HT) neuron processes is distinct from that of the MIP neurons, and no colocalization can be seen. d1, d2 Anterior portion of the optic lobe (frontal view; dorsal is up, lateral to the right). Serotonin-IR (green) cell bodies of medulla (Me) neurons are in two clusters, lying ventrally (vcb) and dorsally (dcb). The three MIP-IR LMIo neuron cell bodies (stars) do not colocalize serotonin but are located close to the vcb cell bodies. Note the serotonergic axons (arrow) destined for the lamina (VLP ventro-lateral protocerebrum). e Main axons of serotonergic (magenta) and MIP-IR (green) neurons destined for the lamina. Note that the axons run closely together (Me medulla). f1, f2 TRH-Gal4 expression (green fluorescent protein, green) in processes distal to the lamina (La) closely matches that of serotonin immunoreactivity (magenta) in double-labeled specimen (JPEG 544 kb)

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High resolution (TIFF 4207 kb)
441_2010_1100_Fig9_ESM.jpg (523 kb)
S. Fig. 2

PDF-immunolabeling (mouse antibody) and MIP-immunolabeling (rabbit anti-MIP) are seen in distinct neurons. a PDF (magenta) expression is seen in lateral clock neurons (s-LNv, l-LNv) and MIP (green) in the three LMIo neurons (LMIo). The boxed area is the ventral part of the accessory medulla (a magnified view is seen in d with altered colors). b1-3 PDF and MIP in area corresponding roughly to the boxed area in a (this is from a different specimen). Processes of LNv (one cell body shown) and LMIo neurons superimpose partly in the region indicated by the arrow (box approximate area corresponding to the accessory medulla in this section). c Relationship between MIP (blue) and PDF (red) neuron processes in the medulla (Me). d Detail of the boxed area in a showing a slight superposition of MIP (blue) and PDF (red) immunolabeled processes in the accessory medulla. e Tim-expressing (green) and MIP-immunolabeled (magenta) neurons in the dorsal brain. Note the lack of MIP expression in DN1 and DN3 clock neurons. Branches of LMIo neurons run dorsally in the dorso-ventral tract (dvt), and the large descending neuron (arrows) also arborizes dorsally near the DN1 neurons. f, g MIP (magenta)- and PDF (green)-expressing neurons showing that LMIo neurons (stars) join the s-LNv axons in the dorso-ventral tract (dvt) to the dorso-lateral protocerebrum (DLP). The large descending neuron (arrows) also runs to the DLP (POT posterior optic tract) (JPEG 788 kb)

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High resolution (TIFF 13644 kb)
441_2010_1100_Fig10_ESM.jpg (545 kb)
S. Fig. 3

Tyrosine hydroxylase (TH)- and tyramine decarboxylase (TDC)-Gal4 expression in relation to MIP immunolabeling (rabbit anti-MIP). a, b Neurons expressing TH-Gal4 (green) are distinct from the MIP-immunolabeled neurons (magenta). Note the LMIo neurons (stars) and cell bodies of some of the TH-expressing optic lobe neurons (arrows). These supply the lobula, but not the medulla or lamina. c, d The MIP-immunolabeled LMIo neurons (stars) do not co-express TDC-Gal4 (green). Two TDC-expressing cell bodies (arrow) supply processes to the medulla (Me) and other parts of the optic lobe (but not lamina) (JPEG 522 kb)

441_2010_1100_MOESM3_ESM.tif (3 mb)
High resolution (TIFF 3122 kb)


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Department of ZoologyStockholm UniversityStockholmSweden

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