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NO/cGMP signalling: L-citrulline and cGMP immunostaining in the central complex of the desert locust Schistocerca gregaria

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Abstract

Nitric oxide (NO) is a gaseous messenger molecule formed during conversion of L-arginine into L-citrulline by the enzyme NO synthase (NOS), which belongs to a group of NADPH diaphorases. Because of its gaseous diffusion properties, NO differs from classical neurotransmitters in that it is not restricted to synaptic terminals. In target cells, NO activates soluble guanylyl cyclase leading to an increase in cGMP levels. In insects, this NO/cGMP-signalling pathway is involved in development, memory formation and processing of visual, olfactory and mechanosensory information. We have analysed the distribution of putative NO donor and target cells in the central complex, a brain area involved in sky-compass orientation, of the locust Schistocerca gregaria by immunostaining for L-citrulline and cGMP. Six types of citrulline-immunostained neurons have been identified including a bilateral pair of hitherto undescribed neurons that connect the lateral accessory lobes with areas anterior to the medial lobes of the mushroom bodies. Three-dimensional reconstructions have revealed the connectivity pattern of a set of 18 immunostained pontine neurons of the central body. All these neurons appear to be a subset of previously mapped NADPH-diaphorase-positive neurons of the central complex. At least three types of central-complex neurons show cGMP immunostaining including a system of novel columnar neurons connecting the upper division of the central body and the lateral triangle of the lateral accessory lobe. Our results provide the morphological basis for further studies of the function of the labelled neurons and new insights into NO/cGMP signalling.

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Abbreviations

BSA:

Bovine serum albumin

CBL:

Lower division of the central body

CBU:

Upper division of the central body

cAMP:

Cyclic adenosine monophosphate

cGMP:

Cyclic guanosine monophosphate

CPU1a:

Columnar (neuron) of the CBU, type 1a

DAB:

3,3'-Diaminobenzidine tetrahydrochloride

DP2:

Dopamine-immunoreactive (neuron) of the pars intercerebralis, type 2

GTP:

Guanosine trisphosphate

IBMX:

3-Isobutyl-1-methylxanthine (inhibitor of phosphodiesterases)

LAL:

Lateral accessory lobe

NADPH:

Reduced form of nicotinamide adenine dinucleotide phosphate

NADPHd:

NADPH diaphorase

NGS:

Normal goat serum

NO:

Nitric oxide

NOS:

Nitric oxide synthase

PB:

Protocerebral bridge

PBS:

Phosphate-buffered saline

PDE:

Phosphodiesterase

POTu:

Posterior optic tubercle

PoU1:

Pontine (neuron) of the CBU, type 1

SNP:

Sodium nitroprusside (nitric oxide donor)

SST:

Saline substituted TRIS buffer

TB2:

Tangential (neuron) of the PB, type 2

TBS:

TRIS-buffered saline

TL1, 2, 4:

Tangential (neurons) of the CBL, types 1, 2, 4

TrX:

Triton X-100

YC-1:

3-[5′-Hyrdroxymethyl-2′-furyl]-1-benzyl indazole (activator of soluble guanylate cyclase)

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Acknowledgements

We thank Dr. Jan de Vente (Maastricht University, The Netherlands) for the generous gift of cGMP antiserum and Pfizer for the gift of sildenafil citrate.

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Authors and Affiliations

  1. Fachbereich Biologie, Tierphysiologie, Philipps Universität Marburg, 35032, Marburg, Germany

    Theresa Siegl, Joachim Schachtner & Uwe Homberg

  2. Departments of Neurology and Neuroscience, Mount Sinai School of Medicine, New York, USA

    Gay R. Holstein

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  1. Theresa Siegl
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  2. Joachim Schachtner
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  3. Gay R. Holstein
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  4. Uwe Homberg
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Correspondence to Uwe Homberg.

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This work was supported by DFG grant HO 950/16-2.

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Siegl, T., Schachtner, J., Holstein, G.R. et al. NO/cGMP signalling: L-citrulline and cGMP immunostaining in the central complex of the desert locust Schistocerca gregaria . Cell Tissue Res 337, 327–340 (2009). https://doi.org/10.1007/s00441-009-0820-z

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