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Polypeptide FFDP and application thereof in oocyte in-vitro culture

A technology for oocyte and in vitro mature culture, which is applied to the biologically active polypeptide FFDP and its application fields, and can solve the problems of imperfect molecular mechanism research and other problems.

Active Publication Date: 2022-01-14
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The research on the molecular mechanism of PCOS oocyte quality decline and ovulation disorder is still incomplete

Method used

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  • Polypeptide FFDP and application thereof in oocyte in-vitro culture
  • Polypeptide FFDP and application thereof in oocyte in-vitro culture
  • Polypeptide FFDP and application thereof in oocyte in-vitro culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Polypeptide FFDP synthesis and analysis of biological parameters

[0033]The sequence of the polypeptide FFDP is Ala Leu Gly Ile Leu Gly Gly Leu Gly Thr Ala Ala, which is a polypeptide molecule consisting of 12 amino acids. The main biological parameters of the peptide obtained through the online peptide analysis tool are as follows: the molecular weight (Molecular weight, Mw) of the peptide is 1013.2 Da, the instability index (Instability Index) is 4.4 hours, the isoelectric point (Theoretical pI) is 5.57, and the fat index ( Aliphatic Index) is 155, is a lipophilic polypeptide (attached figure 1 ). The polypeptide of the sequence shown above was synthesized from the C-terminus to the N-terminus by the Fmoc method of solid-phase synthesis. Its sequence is shown in SEQ ID NO.1.

Embodiment 2

[0034] Example 2. Distribution of FFDP in oocytes

[0035] Synthesize the FITC-labeled transmembrane polypeptide FFDP, that is, insert the FITC tag and the transmembrane sequence at the C-terminus of the polypeptide and synthesize it by solid-phase synthesis Fmoc method. Co-culture FFDP (500nmol) with mouse GV stage oocytes (37°C, 5% CO 2 ), after 14 hours, it was found that the polypeptide was distributed in the cytoplasm by laser confocal microscopy (attached figure 2 ).

Embodiment 3

[0036] Example 3. FFDP promotes in vitro maturation of PCOS model mouse oocytes

[0037] Co-culture FFDP (100 nmol, 500 nml, 1 μmol) with PCOS model mouse oocytes at 37°C, 5% CO 2 ), 3 hours later, the microscopic observation found that the oocyte germinal vesicle rupture rate (GVBD%) of PCOS group mice was observed, and the first polar body excretion rate (Pb1%) was observed 14 hours later. The results showed that with the increase of the peptide concentration, the first polar body exclusion rate of mouse oocytes in the PCOS group increased, but when it was greater than 1 μmol, the maturation rate of the oocytes in the PCOS group decreased, and too much FFDP peptide may affect the oocytes. development of mother cells, so we choose 500nmol as the concentration of FFDP peptide (attached image 3 A-B). , Germinal vesicle rupture (GVBD) is a sign of resuming meiosis of oocytes, GVBD of mouse oocytes in PCOS group (DHEA-treated) was lower than that of control group (Control), af...

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Abstract

The invention discloses a polypeptide FFDP and application thereof in oocyte in-vitro culture. The amino acid sequence of the polypeptide FFDP is as shown in SEQ ID NO. 1. Experiments find that the polypeptide FFDP can be applied to PCOS disease affected oocyte development, and especially applied to the aspect of PCOS oocyte in-vitro maturation culture, or applied to the aspect of improving the meiosis defect during oocyte in-vitro maturation culture.

Description

technical field [0001] The invention belongs to the technical field of assisted reproduction, and in particular relates to a biologically active polypeptide FFDP and its application. Background technique [0002] The problem of fertility disorders has become increasingly prominent, and the number of patients seeking help from assisted reproductive technology (assisted reproduction technology, ART) in clinical outpatient clinics has increased sharply. Although ART is the gospel for infertile people, its overall success rate has always hovered at 40%; it is even lower in patients with polycystic ovary syndrome, ovarian hyperstimulation syndrome, and ovarian secondary hyporesponse. Polycystic ovary syndrome (PCOS) is an endocrine disorder characterized by hyperandrogenism, anovulation or rare ovulation, and possible accompanying polycystic ovaries. It is the main cause of clinical ovulatory infertility . At present, PCOS treatment mainly focuses on lifestyle improvement, and ...

Claims

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Application Information

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IPC IPC(8): C07K14/705C12N5/075
CPCC07K14/70503C12N5/0609C12N2501/50Y02A40/81
Inventor 侯晓静凌钟慧张萍萍郭亚萍季晨博沈嵘
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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