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Kiss-1 / Metastin / GPR54 / AXOR12

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Decreased plasma levels of metastin in early pregnancy are associated with small for gestational age neonates.

OBJECTIVE: To investigate whether pregnancies with small for gestational age (SGA) neonates, defined as customized birth weight below the 10th centile, are associated with altered levels of metastin in maternal plasma in the first trimester.
STUDY DESIGN: Maternal blood was obtained between 8 and 14 weeks of pregnancy. Levels of metastin were measured in pregnancies with (n = 31) or without SGA-neonates (n = 31), matched for gestational age at venipuncture. Measurement of beta-hCG was included to study the influence of gestational age and placental volume on plasma levels of the measured markers.
RESULTS: Metastin was significantly lower in SGA-pregnancies compared to an equal number of matched uneventful pregnancies (metastin: 1376 +/- 1317 pmol/L vs 2035 +/- 1260 pmol/L, p = 0.035; mean +/- standard deviation)(Cat. # : RK-048-56). beta-hCG levels were not different.
CONCLUSION: Metastin is significantly lower in maternal plasma in the first trimester, in pregnancies with SGA-neonates. It might therefore be used in combination with other markers for risk estimation of growth impairment in the first trimester.

Smets et al. Prenat Diagn. 2008 Apr;28(4):299-303.

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GPR54 (KISS1R) Transactivates EGFR to Promote Breast Cancer Cell Invasiveness.

Zajak et al. PLoS One. 2011;6(6):e21599. Epub 2011 Jun 28.
[Note: Article features Phoenix Kisspeptin-10]

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Expression of kisspeptins and kisspeptin receptor in the kidney of chronic renal failure rats.

Shoji et al. Peptides. 2010 Oct;31(10):1920-5. Epub 2010 Jul 17.

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Delayed puberty in spontaneously hypertensive rats involves a primary ovarian failure independent of the hypothalamic KiSS-1/GPR54/GnRH system.

Pinilla et al. Endocrinology. 2009 Jun;150(6):2889-97. Epub 2009 Feb 19.

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Kisspeptin/metastin: a key molecule controlling two modes of gonadotrophin-releasing hormone/luteinising hormone release in female rats.

Uenoyama et al. J Neuroendocrinol. 2009 Mar;21(4):299-304.

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KiSS-1 and GPR54 Genes are Co-Expressed in Rat Gonadotrophs and Differentially Regulated In Vivo by Oestradiol and Gonadotrophin-Releasing Hormone

Richard et al. J Neuroendocrinol. 2008 Mar;20(3):381-93. Epub 2008 Jan 17.

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Developmental and Hormonally Regulated Messenger Ribonucleic Acid Expression of KiSS-1 and its putative receptor GPR54 in Rat Hypothalamus and Potent LH Releasing Activity of KiSS-1 Peptide

Navarro et al. Endocrinology. 2004 Oct;145(10):4565-74. Epub 2004 Jul 8.

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A role for kisspeptins in the regulation of gonadotropin secretion in the mouse

Gottsch et al. Endocrinology. 2004 Sep;145(9):4073-7. Epub 2004 Jun 24

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The GPR54 gene as a regulator of puberty

Seminara et al. N Engl J Med. 2003 Oct 23;349(17):1614-27.

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Dramatic elevation of plasma metastin concentrations in human pregnancy: metastin as a novel placenta-derived hormone in humans

Horikoshi et al. J Clin Endocrinol Metab 2003 Feb;88(2):914-9

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Identification and characterization of mouse metastasis-suppressor KiSS1 and its G-protein-coupled receptor

Stafford et al. Cancer Res 2002 Oct 1;62(19):5399-404

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Metastasis suppressor gene KiSS-1 encodes peptide ligand of a G-protein-coupled receptor

Ohtaki et al. Nature 2001 May 31;411(6837):613-7

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Metastin-like immunoreactivity in the rat medulla oblongata and spinal cord

Dun et al. Neurosci Lett 2003 Jan 2;335(3):197-201

Co-localisation of GPR54 with LH-beta by double immunofluorescence staining from cyclic rat pituitary sections from metoestrous (H-001-64)

Co-localisation of GPR54 with LHbeta

Fluorescent immunohistochemistry was carried out using fluorescein for GPR54 (green) and Texas Red for LHb (red). Dual-labelled cells stand out in yellow ⁄ orange when images are merged (B3). The rare staining for GPR54 was found to be restricted to the anterior lobe (AL, A1). The white arrows show positive cells for GPR54 which are
positive for LHb (B3) and the dark arrows display some LHb cells which are not stained by GPR54 antibody (B2, B3). These immunofluorescent data shows that GPR54 is expressed in gonadotroph cells in the rat. No specific staining was detected in the absence of primary antibody (control-).

Richard et al. J Neuroendocrinol. 2008 Mar;20(3):381-93. Epub 2008 Jan 17.

 

Mapping in Rat Hypothalamus by anti-GPR54 (375-396) Mouse Serum (H-001-64)

Rat hypothalamus was stained by anti-GPR54 (375-396) Mouse Serum

Tissue Sample

Rat hypothalamus tissue

Fixative

10% formalin

Embedding

Paraffin

Negative Control

No primary antibody

Pretreatment

N/A

Blocking

2% Normal Goat Serum

Primary Antibody

Rabbit Anti-GRP54 (375-396) (Mouse) Serum (Catalog No.:H-001-64)

Optimal Dilution

1:50-100 (1hour at RT)

Secondary Antibody

Goat Anti-Rabbit IgG, Biotinylated (1:400), 30 min

Amplification

ABC (Vector) (1:400, 30 min)

Detection System

HRP

Substrate

DAB (Sigma), 3 min

Counterstained

Hematoxylin, 30 sec

AXOR12 (OT7T175, GPR54) is a newly identified orphan G Protein-Coupled Receptor for KiSS-1/Metastin. Mouse pancreas and placenta as well as human placenta tissues can be stained with Phoenix Rabbit anti-AXOR12 (375-398) (Human) antibody (H-048-61).

Mapping in Mouse Pancreas tissue by Anti-AXOR12 (375-398) (Human) Serum (H-048-61)

Mapping in Mouse Pancreatic tissue by Anti AXOR 12 (375-398) (Human) SerumMapping in Mouse Placenta by Anti AXOR 12 Antibody

Tissue Sample

 Mouse Pancreas, Placenta

Fixative

10% formalin

Embedding

Paraffin

Negative Control

No primary antibody

Pretreatment

Intact; Target Retrieval 25 min (Steam)

Blocking

2% Normal Goat Serum

Primary Antibody

Anti-AXOR12 (375-398) (Human) serum (H-048-61)

Optimal Dilution

1: 100

Secondary Antibody

Goat Anti-Rabbit IgG, Biotinylated (1:400), 30 min

Amplification

ABC (Vector)

Detection System

HRP

Substrate

DAB (Sigma), 3 min

Counterstained

Hematoxylin, 30 sec

KiSS-1/Kp-54 and KiSS-1R localization in first trimester placenta (H-048-61)

KiSS-1/Kp-54 and KiSS-1R localization in first trimester placenta.

Sections of human FT placental tissue at week 6-10 of gestation showing the localization of KiSS-1/Kp-54 (A,B) and KiSS-1R (E,F). KiSS-1 and KiSS-1R mRNA were detected by in situ hybridization as dark blue precipitates, whereas their respective proteins were detected using affinity purified polyclonal antibodies evident as dark red precipitate. Sense probes (C,G) and nonimmune sera (D,H) produced no detectable signal. KiSS-1 mRNA (A) and Kp-145/Kp-54 protein (B) were detected mainly on the outer (syncytiotrophoblast) surface of villi. Higher magnification showed that KiSS-1/Kp-54 expression is restricted to the syncytiotrophoblast (insets; A and B; bars, 25 µm), whereas it was undetectable in villous (vCT) and extravillous (evCT) cytotrophoblasts. KiSS-1R mRNA is located in the syncytiotrophoblast (ST), villous (vCT) and extravillous (evCT) cytotrophoblasts (E). This mRNA staining pattern is paralleled by that of KiSS-1R protein (F). Bars, 50 µm.

Bilban et al. J Cell Sci. 2004 Mar 15;117(Pt 8):1319-28.

Mapping in Rat Brain and Spinal Cord by anti metastin serum. (H-048-56)

Tissue Sample

 Rat medulla oblongata and spinal cord

Fixative

4%paraformaldehyde/ 4%picric acid

Embedding

Paraffin

Negative Control

No primary antibody or primary antibody pre-absorbed with the metastin (45-54)-NH2 peptide (10µg/ml) overnight

Pretreatment

Intact

Blocking

2% Normal Goat Serum

Primary Antibody

Metastin (45-54) (Human) Antibody (Code:H-048-56)

Optimal Dilution

1: 1500

Secondary Antibody

Goat Anti-Rabbit IgG, Biotinylated (1:150)

Amplification

ABC (Vector)

Detection System

HRP

Substrate

DAB (Sigma), 3 min

Counterstained

No

Mapping in Rat and Mouse Brain Tissue by Kiss-1 (68-92) / Metastin (1-25) Serum (H-048-62)



Tissue Sample

Rat & Mouse brain tissue

Fixative

10% formalin

Embedding

Paraffin

Negative Control

No primary antibody

Pretreatment

Intact

Blocking

2% Normal Goat Serum

Primary Antibody

Rabbit Anti-Metastin (1-25) / KiSS-1 (68-92) (Human)  Antiserum (Catalog No.:H-048-62)

Optimal Dilution

1:500 (overnight at 4 C)

Secondary Antibody

Goat Anti-Rabbit IgG, Biotinylated (1:400), 30 min

Amplification

ABC (Vector)

Detection System

HRP

Substrate

DAB (Sigma), 3 min

Counterstained

Hematoxylin, 30 sec

Metastin/Kiss-1 EIA Kit (EK-048-59) and Fluorescent EIA Kit (FEK-048-59)

EK-048-59

Metastin/Kiss-1 RIA Kit (RK-048-59)

Peptide
% Cross-reactivity
 KiSS-1 (68-121) Amide / Metastin (1-54) Amide (Human)
 100
 KiSS-1 (112-121)/Metastin (45-54)-NH2 (Human)
 100
 KiSS-1 (68-119) Amide (Mouse)
 0
 KISS-1, Prepro (20-65) (Mouse)
 0
 Metastin (Rat)
 0
 Metastin (26-54)-NH2, [pGlu26] (Human)    
 25
 Neuropeptide AF (NPAF) (Human)
 4
 Neuropeptide FF (NPFF) (Human)
 0
 RFRP-3 (Human)
 0
 FMRF-NH2
 0

 

Kisspeptin-10 EIA (EK-048-56) , Fluorescent EIA (FEK-048-56) & RIA Kit (RK-048-56)

RK-048-56

Peptide
% Cross-reactivity
 KiSS-1 (112-121)/Metastin (45-54)-NH2 (Human)
 100
 KiSS-1 (68-121) Amide / Metastin (1-54) Amide (Human)
 100
 KiSS-1 (68-119) Amide (Mouse)
 0
 KISS-1, Prepro (20-65) (Mouse)
 0
 Metastin (Rat)
 0
 Metastin (26-54)-NH2, [pGlu26] (Human)    
 25
 Neuropeptide AF (NPAF) (Human)
 4
 Neuropeptide FF (NPFF) (Human)
 0
 RFRP-3 (Human)
 0
 FMRF-NH2
 0

Metastin (Rat) RIA kit (RK-048-87)

RK-048-59

Peptide
% Cross-reactivity
 Metastin (Rat) / Kiss-1 (69-119)-amide    
 100
 Kiss-1 (68-119)-Amide (Mouse)
 60
 Metastin (26-54) [pGlu26] Amide (Human)/Kiss-1[pGlu26} (93-121)-Amide
 0
 Metastin (1-54)-Amide (Human) / Kiss-1 (68-121)-Amide
 0
 Metastin (45-54) (Human) / Kiss-1 (112-119)-Amide
 0
 Prepro-Kiss-1 (20-65) (Mouse)
 0

Alignment of human and rat GPR54 amino acid sequences.

Alignment of human and rat GPR54 amino acid sequences.

Identities are indicated, and putative transmembrane domains are boxed. Putative N-glycosylation sites are underlined, and potential phosphorylation sites by PKC are indicated as bold characters.

Kiss-1 (Metastin) Sequence Comparison

Kiss-1 (Metastin) Sequence (Human)

Kiss-1 (Metastin) Prohormone Schematic (Human)

Sequence Comparison between Human & Mouse Kiss-1 Peptide

metastin-small-agonist;Kisspeptin_Antagonist

%001-64%;%048-61%;%metastin%


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