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题名: Molecular Cloning and Differential Expression Pattern of Two Structural Variants of the Crustacean Hyperglycemic Hormone Family from the Mud Crab Scylla Olivacea
作者: Tsai, Kuo-Wei;Chang, Su-Jung;Wu, Hsin-Ju;Shih, Hsin-Yi;Chen, Chun-Hao;Lee, Chi-Ying
贡献者: 生物學系
关键词: Crustacean hyperglycemic hormone;Crustacean hyperglycemic hormone-like;peptide;Tissue expression pattern;CHH family peptide;CHH antisera;Scylla olivacea
日期: 2008-10
上传时间: 2012-07-03T04:02:57Z
出版者: Elsevier
摘要: Two full-length cDNA sequences encoding
a crustacean
hyperglycemic
hormone
(CHH) precursor
were
cloned from tissues
of the mud crab Scylla olivacea.
Sco-CHH (S. olivacea
CHH) was cloned from eyestalk
ganglia,
whereas Sco-CHH-L (S. olivacea
CHH-like peptide)
was cloned from extra-eyestalk
tissues
(pericardial
organ and thoracic
ganglia).
Each conceptually
translated
precursor
is expected to be processed
into a
signal
peptide,
a CHH precursor-
related peptide
(CPRP), and a mature CHH or CHH-like peptide.
The two
precursors
are identical
in amino acid sequence through the 40th residue
of the mature peptide,
but different
from each other substantially
in the C-terminus.
Both CHH variants
contain
the six highly conserved
cysteine
residues
characteristic
of the CHH family
peptides,
and share higher sequence identities
with
other brachyuran
CHH sequences than with those of other taxonomic
groups. As determined
by reverse
transcription-
polymerase
chain reaction
(RT-PCR), the transcripts
of Sco-CHH and Sco-CHH-L were present
in eyestalk
ganglia
and several
extra-eyestalk
tissues
(the thoracic
ganglia,
pericardial
organ, brain,
circumesophageal
connectives,
and gut). Sco-CHH was the predominant
form in eyestalk
ganglia,
while
Sco-CHH-L was the predominant
form in several
extra-eyestalk
tissues.
Neither
transcript
was expressed
in the muscle, hepatopancreas,
ovary, testis,
heart, or gill. Antisera
were raised against synthetic
peptides
corresponding
to a stretch of sequence-specific
to the C-terminus
of Sco-CHH or Sco-CHH-L. Western blot
analyses
of tissues
expressing
Sco-CHH and Sco-CHH-L detected a Sco-CHH immunoreactive
protein
in the
sinus gland, and a Sco-CHH-L immunoreactive
protein
in the pericardial
organ. Immunohistochemical
analyses
of the eyestalk
ganglia
localized
both Sco-CHH and Sco-CHH-L immunoreactivity
to the sinus
gland, and only Sco-CHH immunoreactivity
to the X-organ somata; analyses
of the pericardial
organs also
localized
both Sco-CHH and Sco-CHH-L immunoreactivity
to the anterior
and posterior
bars, as well as to
longitudinal
trunks joining
the two bars. The combined
data provided
supporting
evidence
that Sco-CHH
and Sco-CHH-L are co-localized
in the same tissue.
關聯: General and Comparative Endocrinology, 159(1): 16-25
显示于类别:[生物學系] 期刊論文

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