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stream peptide pathways among lepidopterans (cf. Wise
et al., 2002). Although a previous attempt failed to isolate [His7]-corazonin from B. mori (Hua et al., 2000),
functional [His7]-corazonin may be hidden in Bombyx
brain. This peptide may have some role in neurotransmission between the circadian pacemaker neurons and
peripheral target, as the IHC reactivity showed a dynamic
day/night fluctuation in some neurons and some reactive
neurons share IHC reactivity against DBT.
Fig. 7. Schematic representations of IHC reactive cell bodies
in larval brains. A – cell bodies co-stained with anti-DBT and
anti-[His7]-corazonin sera; B – cell bodies that co-stained with
anti-DBT and anti-[Arg7]-corazonin; C – cell bodies stained
uniquely with anti-[His7]-corazonin; D – cell bodies stained
uniquely with anti-[Arg7]-corazonin. Left hemisphere represent
the larval brain dissected at midday. Right hemisphere the larval
brain dissected at midnight.
Thirdly, unique [His7]-corazonin-like IHC reactivity was
observed.
DISCUSSION
[Arg7]-corazonin was first isolated from P. americana
as a potent cardioaccelerator (Veenstra, 1989) and was
chemically identified from B. mori (Hua et al., 2000). It
induced melanization of body color in Locusta
migratoria, but failed to induce the same effect in B. mori
(Hua et al., 2000). The true function of this peptide in this
species remains unknown, although it has recently been
demonstrated to affect spinning activity when injected
into spinning larvae (Tanaka et al., 2002). The present
study suggests that its secretion may be under circadian
control, as it is co-localized with PER, as in M. sexta.
Not only [Arg7]-corazonin, but also [His7]-corazoninlike IHC reactivities, were present in the BR-SG complex
of B. mori. The reactivities against both corazonins partially overlapped but not always (Fig. 7). The shared reactivities against both DBT and [His7]-corazonin occurred
in some neurons, but these neurons were not the same
ones that showed shared reactivities against DBT and
[Arg7]-corazonin (Fig. 7A, B). This may show that circadian output pathways are not singular, but that there are
two distinct corazonin output pathways, even in the same
species. Day/night variation in IHC reactivity was
apparent in different neurons.
Distinct patterns of distribution of the two corazonins,
although partially overlapping, were obtained in the present investigation. Together with the cross-reactivity
shown in Table 1, both [His7]- and [Arg7]-corazonin-like
antigens are co-localized in some clock neurons, though
antisera may cross-react with different corazonin
antigens. The results indicate at least a diversity in down-
286
ACKNOWLEDGEMENTS. We thank Dr. J.A. Veenstra for
antisera against [Arg7]-corazonin and Dr. Kazuei Mita for an
EST clone of DBT in B. mori. We dedicate this paper to the
memory of Dr. Naoyuki Ichihara who produced antiserum
against DBT. The following grant supports are appreciated from
JSPS (11460021 from Grant-in-Aid for Scientific Research B
and 99L01205 from the “Research for Future” Program).
REFERENCES
HELFRICH-FORSTER C. 1995: The period clock gene is expressed
in central nervous system neurons which also produce a neuropeptide that reveals the projections of circadian pacemaker
cells within the brain of Drosophila melanogaster. Proc. Nat.
Acad. Sci. USA 92: 612–616.
HOMBERG U., DAVID N.T. & HILDEBRAND J.G. 1991: Peptideimmunocytochemistry of neurosectory cells in the brain and
retrocerebral complex of the sphix moth Manduca sexta. J.
Comp. Neurol. 303: 35–52.
HUA Y.J., ISHIBASHI J., SATOP H., TAWFIK A.L., SAKAKIBARA M.,
TANAKA Y., DERUA R., WAELKENS E., BAGGERMAN G., DE LOOF
A., SCHOOFS L. & TANAKA S. 2000: Identification of
[Arg7]-corazonin in the silkworm, Bombyx mori and the
cricket, Gryllus bimaculatus as a factor inducing dark color in
an albino strain of the locust, Locusta migratoria. J. Insect
Physiol. 46: 853–859.
RENN S.C.P., ROSBASH M., HALL J.C. & TAGHERT P. H. 1999: A
pdf neuropeptide gene mutation and ablation of PDF neurons
each cause severe abnormalities of behavioral circadian
rhythms in Drosophila. Cell 99: 791–802.
SAUMAN I. & REPPERT S.M. 1996: Circadian clock neurons in the
silkmoth, Antheraea pernyi; novel mechanisms of period protein regulation. Neuron 117: 889–900.
TANAKA Y., HUA Y-J., ROLLER L. & TANAKA S. 2002: Corazonin
reduces the spinning rate in the silkworm, Bombyx mori. J.
Insect Physiol. 48: 743–749.
VEENSTRA J.A. 1989: Isolation and structure of corazonin, a cardioactive peptide from American cockroach. FEBS Lett. 250:
231–234
VEENSTRA J.A. 1991: Presence of corazonin in three insect species, and isolation and identification of [His7]-corazonin from
Schistocerca americana. Peptides 12: 1285–1289
WILLIAMS J.A. & SEHGAL A. 2001: Molecular components of the
circadian system in Drosophila. Annu. Rev. Physiol. 63:
729–755.
WISE S., DAVIS N.T., TYNDALE E., NOVERAL J., FOLWELL M.G.,
BEDIAN V., EMERY I.F. & SIWICKI K.K. 2002: Neuronanatomical studies of period gene expression in the hawkmoth,
Manduca sexta. J. Comp. Neurol. 447: 366–380.
Received October 7, 2002; revised January 6, 2003; accepted May 6, 2003