Synthesis of a novel peptide, galanin-like peptide (GALP), by a combination of recombinant DNA technology and chemical cleavage reactions

Itoh, Takashi; Miwa, Masanori; Suenaga, Masato; Ohtaki, Tetsuya; Kitada, Chieko; Nishimura, Osamu; Fujino, Masahiko

doi:10.1039/b001087l

Cited by 9 publications

(7 citation statements)

References 11 publications

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“…The icv administration of GALP was performed as described previously (11). Briefly, recombinant rat GALP (17) was dissolved in distilled water at 1 mm, diluted twice with 2-fold concentrated PBS, and further diluted 5-fold with PBS. Ten microliters of GALP solution (0.5 or 0.1 mm) or vehicle (PBS) were administered into the third ventricle of male Zucker obese and lean rats (11 wk old, n ϭ 5/group) through steel cannulas during the early light phase (0900 -1100 h).…”

Section: The Icv Administration Of Galpmentioning

confidence: 99%

Changes in Hypothalamic Expression Levels of Galanin-Like Peptide in Rat and Mouse Models Support That It Is a Leptin-Target Peptide

et al. 2003

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Galanin-like peptide (GALP) is a novel peptide that has been isolated from the porcine hypothalamus. The expression of GALP mRNA is localized to the hypothalamic arcuate nucleus and is thought to be under the regulation of leptin. First, we confirmed by real-time PCR analysis that sc administration of leptin to Wistar rats under food-deprived conditions resulted in a 1.5-fold increase in hypothalamic GALP mRNA levels. Next, GALP mRNA levels were found to be reduced by 50% in 11-wk-old male Zucker obese rats compared with age-matched Zucker lean rats, whereas neuropeptide Y mRNA levels were increased by 55% and proopiomelanocortin mRNA levels were reduced by 53% in Zucker obese rats. Analysis using a two-site enzyme immunoassay revealed a lower level of hypothalamic GALP immunoreactivity in 11-wk-old Zucker obese rats (5.9 fmol/mg protein) than in age-matched Zucker lean rats (19.6 fmol/mg protein). Immunohistochemical studies demonstrated that Zucker obese rats (11 wk old) had a reduced number of GALP immunoreactivity-positive cells (29.4 cells/3 slices) in the arcuate nucleus compared with age-matched Zucker lean rats (115 cells/3 slices). Furthermore, Zucker obese rats showed increased sensitivity to intracerebroventricularly administered GALP compared with Zucker lean rats, in that a lower dose of GALP increased plasma LH levels in male Zucker obese rats, but not in male Zucker lean rats. In addition, a reduction in the level of hypothalamic GALP mRNA was found in db/db and ob/ob mice. The result supports the hypothesis that the hypothalamic GALP gene expression is controlled by leptin signals and suggests possible involvement of GALP in the reproductive abnormalities of the Zucker obese rat.

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Section: The Icv Administration Of Galpmentioning

confidence: 99%

Changes in Hypothalamic Expression Levels of Galanin-Like Peptide in Rat and Mouse Models Support That It Is a Leptin-Target Peptide

et al. 2003

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“…Rat GALP was synthesised as described previously (18). The peptide was dissolved in ultra‐pure water at 1 m m and stored at −80 °C.…”

Section: Methodsmentioning

confidence: 99%

Oestrogen‐Dependent Stimulation of Luteinising Hormone Release by Galanin‐Like Peptide in Female Rats

Uenoyama

Tsukamura

Kinoshita

et al. 2008

J Neuroendocrinology

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Galanin-like peptide (GALP), a ligand for three types of galanin receptor, is reported to have a role in regulating luteinising hormone (LH) release in male rodents and primates, but its role in LH release in female rodents remains controversial. The present study was conducted to test whether GALP has a stimulatory role in regulating LH secretion in female rats. The effect of i.c.v. infusion of GALP (5 nmol) on pulsatile LH release was investigated in Wistar-Imamichi strain female rats, or lean and obese Zucker rats. In oestradiol-17beta (oestradiol)-primed ovariectomised (OVX) Wistar-Imamichi female rats, i.c.v. infusion of GALP caused a gradual increase in LH release for the first 1.5 h after the infusion followed by an increased LH pulse frequency during the next 1.5 h, resulting in a significant increase in the mean LH concentrations and baseline levels of LH pulses throughout the sampling period and in the frequency of LH pulses at the last half of the period compared to vehicle-treated controls. The stimulatory effect of GALP was oestrogen-dependent because the same GALP treatment did not affect LH release in OVX rats in the absence of oestradiol. In lean Zucker rats, LH pulses were found in oestradiol-primed OVX individuals and central GALP infusion increased mean LH concentrations in the last half of the period. By contrast, few LH pulses were found in oestradiol-primed OVX obese Zucker rats reportedly with lower hypothalamic GALP expression. Central GALP infusion caused an apparent but transient increase in LH release, resulting in the significant increase in all pulse parameters of LH pulses compared to vehicle-treated controls in the first half of the sampling period. These results suggest that hypothalamic GALP is likely involved in stimulating GnRH/LH release, and that the stimulatory effect of GALP on LH release is oestrogen-dependent in female rats.

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“…Lyophilised recombinant porcine GALP (4.3 mg) was generously donated by Dr. Tetsuya Ohtaki from Takeda Chemical Industries, Japan (Ito et al, 2000).…”

Section: Methodsmentioning

confidence: 99%

Presence of transient helical segments in the galanin-like peptide evident from 1H NMR, circular dichroism, and prediction studies

Dastmalchi

Church

Morris

et al. 2004

Journal of Structural Biology

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Synthesis of a novel peptide, galanin-like peptide (GALP), by a combination of recombinant DNA technology and chemical cleavage reactions

Abstract: Galanin-like peptide, a newly isolated peptide ligand for the GALR2 receptor, was synthesized by a combination of recombinant DNA technology and chemical cleavage reactions.

Cited by 9 publications

References 11 publications

Changes in Hypothalamic Expression Levels of Galanin-Like Peptide in Rat and Mouse Models Support That It Is a Leptin-Target Peptide

Changes in Hypothalamic Expression Levels of Galanin-Like Peptide in Rat and Mouse Models Support That It Is a Leptin-Target Peptide

Oestrogen‐Dependent Stimulation of Luteinising Hormone Release by Galanin‐Like Peptide in Female Rats

Presence of transient helical segments in the galanin-like peptide evident from 1H NMR, circular dichroism, and prediction studies

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