Investigation into the presence and functional significance of proinsulin C-peptide in the female germline†

Martin, Jacinta H.; Aitken, R. John; Bromfield, Elizabeth G.; Cafe, Shenae L.; Sutherland, Jessie M.; Frost, Emily R; Nixon, Brett; Lord, Tessa

doi:10.1093/biolre/ioz008

Cited by 6 publications

(5 citation statements)

References 90 publications

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“…However, a recent study suggested the presence of an alternative C-peptide receptor instead of GPR146; GPR146 has not been confirmed as a C-peptide receptor when GPR146 was analyzed using dynamic mass distribution, an arrestin binding assay, and cellular binding and internalization under a fluorescent microscope (Lindfors et al, 2020). Meanwhile, C-peptide regulates the function of downstream effectors via direct binding, and binds to the breast cancer type 2 susceptibility protein (BRAC2) in mammalian zygotes in response to fertilization stimulus, and causes changes in the subcellular localization of BRAC2 to the nucleus, inducing aberrant meiotic resumption and perturbation of the DNA repair system (Martin et al, 2019). C-peptide might also directly bind to and activate the downstream effector proteins Akt or PP1.…”

Section: Discussionmentioning

confidence: 99%

C-Peptide Promotes Cell Migration by Controlling Matrix Metallopeptidase-9 Activity Through Direct Regulation of β-Catenin in Human Endometrial Stromal Cells

Khaliq

Umair

Baek

et al. 2022

Front. Cell Dev. Biol.

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The motility of endometrial stromal cells (ESCs) contributes to the restoration of the endometrial functional layer and subsequently supports the trophoblast invasion during early pregnancy. Following ESCs differentiation through decidualization in response to progesterone during the menstrual cycle and embryo implantation, decidualized ESCs (D-ESCs) have greater motility and invasive activity. The human proinsulin-connecting peptide (C-peptide) is produced in equimolar amounts during the proteolysis of insulin in pancreatic β-cells. However, the function of C-peptide in the cellular motility of the human endometrium remains unexamined. In the present study, C-peptide was identified as a determinant of undecidualized human endometrial stromal cells (UnD-ESCs) migration. C-peptide promoted the migration and invasion of UnD-ESCs and trophoblast-derived Jeg3 cells, but not that of ESCs post decidualization, a functional and biochemical differentiation of UnD-ESCs. Both Akt and protein phosphatase 1 regulated β-catenin phosphorylation in UnD-ESCs, not D-ESCs, thereby promoting β-catenin nuclear translocation in C-peptide-treated UnD-ESCs. C-peptide was also observed to increase matrix metallopeptidase-9 (MMP9) activity by increasing MMP9 expression and decreasing the expression of metallopeptidase inhibitor 1 (TIMP1) and TIMP3. Their expression was modulated by the direct binding of β-catenin in the regulatory region of the promoter of MMP9, TIMP1, and TIMP3. Inhibition of either β-catenin or MMP9 dampened C-peptide-enhanced migration in UnD-ESCs. Together, these findings suggest that C-peptide levels are critical for the regulation of UnD-ESC migration, providing evidence for the association between C-peptide levels and the failure rate of trophoblast invasion by inducing abnormal migration in UnD-ESCs in hyperinsulinemia or PCOS patients.

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Section: Discussionmentioning

confidence: 99%

C-Peptide Promotes Cell Migration by Controlling Matrix Metallopeptidase-9 Activity Through Direct Regulation of β-Catenin in Human Endometrial Stromal Cells

Khaliq

Umair

Baek

et al. 2022

Front. Cell Dev. Biol.

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“…Immunofluorescence analysis was performed on Bouin’s fixed ovary sections, as described previously 59 . Sections were de-paraffinised and rehydrated via a series of xylene and ethanol washes (100%, 75% and 0% ethanol in H 2 O), respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Superovulation was conducted as described previously 59 , 60 . 4–6 week old females were subjected to an injection regimen of equine Chorionic Gonadotropin (eCG) (Intervet, Sydney, Australia), followed by human Chorionic Gonadotropin (hCG) (Intervet), 48 h later.…”

Section: Methodsmentioning

confidence: 99%

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EPAS1 expression contributes to maintenance of the primordial follicle pool in the mouse ovary

Martin,

Bernstein,

Lyons

et al. 2024

Sci Rep

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Oxygen availability can have profound effects on cell fate decisions and survival, in part by regulating expression of hypoxia-inducible factors (HIFs). In the ovary, HIF expression has been characterised in granulosa cells, however, any requirement in oocytes remains relatively undefined. Here we developed a Hif2a/Epas1 germline-specific knockout mouse line in which females were fertile, however produced 40% fewer pups than controls. No defects in follicle development were detected, and quality of MII oocytes was normal, as per assessments of viability, intracellular reactive oxygen species, and spindle parameters. However, a significant diminishment of the primordial follicle pool was evident in cKO females that was attributed to accelerated follicle loss from postnatal day 6 onwards, potentially via disruption of the autophagy pathway. These data demonstrate the importance of HIF signalling in oocytes, particularly at the primordial follicle stage, and lend to the importance of controlling oxygen tension in the development of in vitro growth and maturation approaches for assisted reproduction.

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“…Super ovulated females were then housed overnight with male C57/BL6/CBA-F1 mice for mating (2 females/male) as summarised in Figure 1 . The following morning, females were euthanised by cervical dislocation and presumptive zygotes were collected from the oviducts and washed in G-1 plus media (Vitrolife, Gothenburg Sweden) ( Martin et al, 2016 ; Martin et al, 2018 ; Martin et al, 2019 ). Presumptive zygotes were then treated with 300 μg/ml hyaluronic acid (Sigma-Aldrich, Missouri United States) for 30–45 s to remove the outer cumulus cells.…”

Section: Methodsmentioning

confidence: 99%

Placental deficiency of the (pro)renin receptor ((P)RR) reduces placental development and functional capacity

Schofield,

Kahl,

Rodrigues

et al. 2023

Front. Cell Dev. Biol.

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The (pro)renin receptor ((P)RR; also known as ATP6AP2) is a multifunctional receptor. The (P)RR activates the tissue renin-angiotensin system (RAS) and is also involved in regulating integral intracellular pathways such as V-ATPase and Wnt/β-catenin signalling. Given this, the (P)RR may be associated with essential pathways in placentation, however its role within the context of pregnancy remains poorly characterised. The first trimester/extravillous trophoblast cell line, HTR-8/SVneo, underwent an siRNA knockdown where they were incubated for 24 h with a negative control siRNA or siRNA targeting ATP6AP2 mRNA. xCELLigence real-time cell analysis was performed to assess the effect of ATP6AP2 mRNA knockdown on HTR-8/SVneo cell proliferation, migration, and invasion. In subsequent experiments, GFP-encoding lentiviral packaged gene-constructs were used to knockdown (P)RR expression in the trophectoderm of C57/BL6/CBA-F1 mouse blastocysts. Blastocysts were incubated for 6 h with vehicle (no-virus), control virus (non-targeting shRNA and GFP), or (P)RR-knockdown virus ((P)RR shRNA and GFP) before transfer into recipient pseudo-pregnant Swiss CD1 female mice. Fetal and placental tissues were collected and assessed at embryonic age (EA) 10 and 18. (P)RR levels were measured in the labyrinth zone of day 18 placentae and stereological Merz grid analysis was performed to determine the volumetric distribution of trophoblasts, fetal capillaries, and the maternal blood space. We showed that a reduction of ATP6AP2 expression in HTR-8/SVneo cells in vitro, impaired trophoblast proliferation, migration, and invasion. In vivo, decreasing placental labyrinth (P)RR expression adversely effected placental physiology, decreasing placental trophoblast number and total surface area available for exchange, while also increasing maternal blood space. Additionally, decreased (P)RR affected placental efficacy evident by the reduced fetal-placental weight ratio. Our study shows that the (P)RR is necessary for appropriate placental development and function.

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Investigation into the presence and functional significance of proinsulin C-peptide in the female germline†

Cited by 6 publications

References 90 publications

C-Peptide Promotes Cell Migration by Controlling Matrix Metallopeptidase-9 Activity Through Direct Regulation of β-Catenin in Human Endometrial Stromal Cells

C-Peptide Promotes Cell Migration by Controlling Matrix Metallopeptidase-9 Activity Through Direct Regulation of β-Catenin in Human Endometrial Stromal Cells

EPAS1 expression contributes to maintenance of the primordial follicle pool in the mouse ovary

Placental deficiency of the (pro)renin receptor ((P)RR) reduces placental development and functional capacity

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