Identification of the cerebral neurosecretory cells that contain eclosion hormone in the moth Manduca sexta

Copenhaver, PF; Truman, JW

doi:10.1523/jneurosci.06-06-01738.1986

Cited by 79 publications

(40 citation statements)

References 26 publications

(36 reference statements)

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“…EH immunohistochemistry was carried out using a rabbit polyclonal antibody raised against partially purified Manduca sexta EH [35]. For use on Drosophila tissue, we preadsorbed it with 0-12-h Drosophila embryos.…”

Section: Immunocytochemistrymentioning

confidence: 99%

“…The location of the pair of neurons identified by in situ hybridization corresponds to a pair of brain cells that are immunopositive to an antiserum raised against Munducu EH [35]. In the larval stage these cells possess a bifurcating axon, one branch of which extends peripherally to the corpora cardiaca portion of the ring gland (Fig.…”

Section: Expression Of the Eh Genementioning

confidence: 99%

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Isolation, characterization and expression of the eclosion hormone gene of Drosophila melanogaster

Horodyski

Ewer

Riddiford

et al. 1993

European Journal of Biochemistry

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Eclosion hormone (EH) is a neuropeptide that triggers the performance of ecdysis behaviors at the end of a molt. We have isolated the EH gene from Drosophila melanogasfer, and localized the gene to the right arm of chromosome 3 at band position 90B1-2. The 97-amino-acid translation product contains a signal peptide followed by a 73-amino-acid prohormone. The N-terminus of the prohormone has diverged from lepidopteran EH both in its length and amino acid composition, and contains a potential endoproteolytic cleavage site. The deduced sequence of Drosophila EH is 58% identical (36 of 62 amino acids) to that of Manduca EH. The EH gene is expressed as a 0.8-kb transcript in a single pair of brain neurons which extend their processes the entire length of the central nervous system and also to the corpora cardiaca portion of the ring gland. These cells show massive depletion of immunoreactive EH at ecdysis.Over the last 10 years, work in invertebrates in general and insects in particular has led to identification of a large number of neuropeptides, many of which are expressed in a small number of cells within the central nervous system (CNS) [l, 21. These findings raise a number of important questions such as the role of these peptides in the normal physiology and development of the organism and the mechanisms that restrict peptide expression to a select set of cells within the CNS. To answer both questions, Drosophila melunogaster provides a number of distinct advantages. Mutants in a desired gene can be generated with relative ease and P-element transformation technology allows examination of regulatory regions that control peptide expression [3, 41. Consequently, we have turned to Drosophila to further our analysis of the neuropeptide eclosion hormone (EH), that triggers ecdysis of insects [5].Eclosion hormone was first found in moths [6] and later shown to be a 62-amino-acid peptide [7 -91 produced by two pairs of ventromedial neurosecretory cells in the brain [lO-131. In moths it has marked behavioral and developmental actions. It is known to act directly on the CNS to release the stereotyped motor programs that bring about the shedding of the old cuticle at the end of each molt [14]. Circulating peptide also acts on peripheral tissues to cause a diverse set of effects such as an increase in plasticity of wing cuticle [15], discharge of secretion by dermal glands [16], and the programmed degeneration of ecdysial muscles [17]. Yet the full range of EH actions are not known because it is not possible to block selectively either release or action of EH in moths. It appeared feasible to study EH in Drosophila because CNS extracts of both Calliphora [5] and Drosophila (Kimura and Truman, unpublished data) show EH activity in moth bioassays. Moreover, blood from ecdysing fleshflies, Sarcophaga bullata, stimulates premature ecdysis behavior in pharate (before ecdysis) flies [18]. The recent cloning of the EH gene from two species of moths, Munduca sexfa [ll] and Bombyx mori [13], has provided an avenue to search for t...

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Section: Immunocytochemistrymentioning

confidence: 99%

Section: Expression Of the Eh Genementioning

confidence: 99%

Isolation, characterization and expression of the eclosion hormone gene of Drosophila melanogaster

Horodyski

Ewer

Riddiford

et al. 1993

European Journal of Biochemistry

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“…SC). No obvious hybridization was seen in the lateral neurosecretory cell cluster thought to be the source of EH for adult eclosion (32).…”

Section: Resultsmentioning

confidence: 91%

“…4B), we constructed a cDNA library from day-9 to -12 developing adult brain mRNA in AZAP II (Stratagene). The EH mRNA is likely only found in a few neurosecretory cells in the brain (32) and, therefore, is rare [<0.01% of the poly(A)+ RNA population], even in the portion of the brain enriched for these cells. To facilitate screening, we amplified phage DNA from aliquots of the cDNA library between two oligonucleotides derived from the sequence of the EH genomic clone AD39 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Isolation and expression of the eclosion hormone gene from the tobacco hornworm, Manduca sexta.

Horodyski

Riddiford²,

Truman³

1989

Proc. Natl. Acad. Sci. U.S.A.

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Eclosion hormone (EH) is a 62-amino acid neuropeptide that initiates the ecdysis behavior of insects. The EH-encoding gene of the tobacco hornworm, Manduca sexta, was isolated by using a designed 72-mer oligonucleotide probe. Sequence analysis of this gene and its corresponding cDNA showed that the EH gene is 7.8 kilobases and consists of three exons. Exon I is totally nontranslated; exon II contains a 26-amino acid signal peptide and amino acids 1-4 of the EH peptide, and exon III encodes the remainder of the peptide. The EH gene is present in a single copy per haploid genome and transcribes an 0.8-kb mRNA that is expressed in larval, diapausing pupal, and developing adult brains but not in the ventral nerve cord or in nonneural tissues. In situ hybridization showed that the EH gene is expressed in two pairs of ventromedial neurosecretory cells in brains of both larvae and developing adults.As in vertebrates, insect neuropeptides are a diverse class of regulatory molecules that affect a number of physiological processes relating to growth, metamorphosis, reproduction, and homeostasis (1, 2). Despite their importance, the structures of relatively few of these neuropeptides are known; even less is known about the genes that code for them. Recent advances have been made in the molecular characterization and cloning of genes for the insect neuropeptides PheMet-Arg-Phe-NH2 (FMRFamide) (3, 4), bombyxin (5), and adipokinetic hormone (6).Growth and metamorphosis in insects is characterized by a series of molts when a new cuticle is produced. The shedding of the old cuticle at ecdysis is caused by the eclosion hormone (EH), a neuropeptide that triggers this behavior and directs associated physiological changes (7). The primary target of this peptide is the central nervous system, although other nonneural targets are known. Purification and sequencing of EH from two different moths, Manduca sexta (8, 9) and Bombyx mori (10), has revealed a 62-amino acid peptide that shows 80% identity between the two species. This neuropeptide, however, has no similarity with any other known neuropeptide from either vertebrates or invertebrates (8).Using an oligonucleotide based on this sequence, we now report the isolation and characterization of the EH genet.This gene proves to code only for the pre-EH molecule and to be expressed primarily in a set of four ventromedial cells in the brain of larvae and of developing adults.EXPERIMENTAL PROCEDURES Animals. Larvae of the tobacco hornworm, M. sexta, were reared individually on an artificial diet at 26°C under a 17-hr light:7-hr dark photoperiod as described (11). At the start of the wandering stage, the larvae were transferred to wooden blocks in which they pupated. Developing adults were transferred to a photoperiod of 12-hr light: 12-hr dark. Staging of developing adults was by days after pupal ecdysis, except for the final 5 days, which were determined by reference to a defined set of cuticular markers (12).Oligonucleotide Probe Construction and Labeling. To construct the 72 nu...

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“…Two of them send their axons to the contralateral CC (8); originally one of these cells was suggested to be prothoracicotropic ( 1 ) but later both of them were shown to react with antibody to the "big" PTTH (32). The remainder of the LNC group in Manduca reportedly consists of as many as 10 cells supplying axons to the ipsilateral CC (8); these LNC probably include the 5 cells that were identified as the source of the eclosion hormone (10). Some of these cells in adult Manduca contain insu I i n-l i ke immunoreact ivity (11).…”

Section: Localization Of Neurons Containing Bombyxin-like Materialsmentioning

confidence: 99%

Developmental Changes in the Bombyxin‐and Insulin‐like Immunoreactive Neurosecretory System in the Wax Moth, Galleria mellonella

Žitňan¹,

Sehnal

Mizoguchi

et al. 1990

Dev Growth Differ

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Four medial neurosecretory cells (MNC) and 4 lateral neurosecretory cells (LNC) in each brainhemisphere, and one pair of cells in each thoracic ganglion (TG) of Galleria larva react with antibodies against bombyxin and insulin. Material secreted from the MNC and LNC is released mainly in the corpora allata, and that from the TG through the ventral median nerves. Intrinsic secretory cells of the corpora cardiaca (CC) also contain bombyxin-like, but not insulin-like material. The immunoreactivities all disappear during molts and reappear with resumption of feeding. In the MNC and TG they reappear for less than a day, but in cells of the CC immunoreactivity reappears for the whole feeding period. Before pupation, the LNC become temporarily immunopositive towards the end of feeding period, and the MNC and TG during the wandering period, i.e. at the time of prothoracic gland stimulation. lmmunoreactivity disappears during the pupal molt. In pupae it is present in the 4 pairs of MNC and 1-2 pairs of LNC 12-48 hr after ecdysis, and in cells of the CC from 12 hr after ecdysis until the end of the pupal instar. In adult, immunoreactivity is restricted to 2 pairs of the LNC and to CC cells.

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Identification of the cerebral neurosecretory cells that contain eclosion hormone in the moth Manduca sexta

Cited by 79 publications

References 26 publications

Isolation, characterization and expression of the eclosion hormone gene of Drosophila melanogaster

Isolation, characterization and expression of the eclosion hormone gene of Drosophila melanogaster

Isolation and expression of the eclosion hormone gene from the tobacco hornworm, Manduca sexta.

Developmental Changes in the Bombyxin‐and Insulin‐like Immunoreactive Neurosecretory System in the Wax Moth, Galleria mellonella

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