Dataset Information

Characterization of serum and tissue oxytocinase and tissue oxytocin in the pregnant and nonpregnant mare

ABSTRACT: Leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase) is a key regulator of oxytocin. Our objectives were to measure serum concentrations of LNPEP and determine the presence of LNPEP and oxytocin (OXT) in equine tissue. LNPEP was measured by ELISA in serum obtained: every other day from ovulation (D0) until D21-22 in nonpregnant and pregnant mares; and at around 320 days of gestation, at 24 hours before foaling, and 20 min and 2 hrs post-foaling. Placenta from the pregnant horn (PH), nonpregnant horn (NPH) and body (B) and from tissues of 6 nonpregnant mares were homogenized and extracts obtained for protein determination and LNPEP ELISA assay. Identification of LNPEP and OXT protein in tissues was also performed via western blot (WB), immunohistochemistry (IHC) and liquid chromatography-mass spectrometry (LC-MS/MS). Furthermore, in situ hybridization was performed for LNPEP and OXT on endometrium, myometrium, pituitary and corpus luteum (CL). No difference in serum LNPEP concentration was identified between any of the groups. The NPH had the lowest quantities of LNPEP (P<0.05). The highest to lowest LNPEP U/mg protein by tissue were: myometrium> follicle wall> endometrium> kidney>CL> liver. Oxytocin was identified in the equine pituitary, CL and placenta. LNPEP may regulate the availability of luteal OXT locally.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Equus Caballus (horse)

TISSUE(S): Embryo, Myometrium, Placenta, Liver, Uterine Endometrium, Corpus Luteum, Neurohypophysis

SUBMITTER: Mariana Diel de Amorim

LAB HEAD: Mariana Diel de Amorim

PROVIDER: PXD036368 | Pride | 2023-05-10

REPOSITORIES: Pride

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Publications

Characterization of serum and tissue oxytocinase and tissue oxytocin in the pregnant and non-pregnant mare.

Diel de Amorim Mariana M Dong Lynn L Byron Michael M Foster Robert A RA Klein Claudia C Saleh Monique M Saleh Tarek T Card Claire C

Scientific reports 20230321 1

Oxytocin is a hormone with functions in: reproduction, maternal bonding, milk ejection, and feeding/social behavior, and is reported to be present in a variety of tissues. Our goal is to characterize oxytocin and leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase), a key regulator of oxytocin in mares. We measured serum and tissue LNPEP by ELISA from ovulation (D0) until D21-22 in non-pregnant (n = 5) and pregnant mares (n = 6); and in periparturient and postpartum mares (n = 18). Placenta (n ...[more]

PMID: 36944665

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Project description:The equine endometrium exhibits characteristic morphological and functional changes during the estrous cycle controlled by the interplay of progesterone and estradiol. A microarray analysis of endometrial tissue samples derived from 5 time points of the estrous cycle (D0, D3, D8, D12, and D16) was performed to study the dynamics of endometrial gene expression. Endometrial biopsies were collected from five mares (Bavarian Warmblood) at the respective time points. Samples were divided and subjected to isolation of RNA for microarray analysis and analysis of tissue composition. Blood samples were collected to determine serum progesterone levels for every sample. Statistical analysis of microarray data revealed almost 10,000 differential probes corresponding to 4,996 differentially expressed genes. A cluster analysis based on gene expression profiles during the estrous cycle revealed 8 major gene expression profiles: mRNAs with highest levels 1) at D0, 2) from D0 to D3, 3) at D3, 4) from D3 to D8, 5) at D8, 6) from D8 to D12, 7) from D12 to D16, and 8) at D16. DAVID Functional Annotation Clustering revealed overrepresentation of distinct functional terms in different phases of the cycle, e.g. M-bM-^@M-^Xextracellular matrixM-bM-^@M-^Y and M-bM-^@M-^Xprotein transportM-bM-^@M-^Y during estrus, M-bM-^@M-^XDNA replication and M-bM-^@M-^Xcell cycleM-bM-^@M-^Y during early luteal phase, M-bM-^@M-^Xendoplasmic reticulumM-bM-^@M-^Y and M-bM-^@M-^Xprotein transportM-bM-^@M-^Y in the luteal phase, and M-bM-^@M-^Xinflammatory responseM-bM-^@M-^Y in the late luteal and follicular phase. Expression of selected genes of the expression clusters was validated by quantitative Real-time PCR (qPCR). This study provides new insights into global changes of equine endometrial gene expression during the estrous cycle. Equine endometrial tissue samples were collected at 5 time points during the sexual (estrous) cycle from 5 mares (5 biological replicates per time point) and analyzed with Agilent microarrays.

Dataset Information

Characterization of serum and tissue oxytocinase and tissue oxytocin in the pregnant and nonpregnant mare

Publications

Characterization of serum and tissue oxytocinase and tissue oxytocin in the pregnant and non-pregnant mare.

Similar Datasets

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