Development of an Enzyme Immunoassay for Urinary Pregnanediol-3-Glucuronide in a Female Giant Panda (Ailuropoda melanoleuca)

Abstract

In order to enable monitoring of the reproductive status of the female giant panda after observation of estrus behavior, we developed an enzyme immunoassay (EIA) system for urinary pregnanediol-3-glucuronide (PdG), a progesterone metabolite, using commercial reagents and examined the changes in the urinary concentration of PdG in a female giant panda that showed pseudopregnancy and suspicious pseudopregnancy in 6 consecutive years. The developed EIA system had good reproducibility (intra-and interassay CVs 6.1% and 16.3%, respectively), good parallelism between the standard curve and the dose response curve of serial diluted samples and positive correlation (r=0.836) with the data for PdG in the same samples measured by gas chromatography. Urinary PdG in the female panda showed two phases of increase. The first elevation was observed immediately after estrus with the levels of PdG below 100 ng/ Crmg, while the second phase was characterized by a drastic elevation above 100 ng/Crmg until the level began to decrease at the end of pseudopregnancy or suspicious pseudopregnancy. The length of the second phase had wider range than that of the first phase. In the present study, a new EIA assay system for urinary PdG in the female giant panda was developed, and we found that the length of the second phase is unstable in the pseudopregnant and suspicious pseudopregnant giant panda, in contrast with the unstable length of the first phase caused by delayed implantation in the pregnant giant panda.

Urinary sex steroid hormone and placental leucine aminopeptidase concentration differences between live births and stillbirth of Bornean orangutans (Pongo pygmaeus)

“…Urine concentrations of E 1 G and PdG were assayed using an enzyme immunoassay using the same methods and reagents as described by Hama et al . The polyclonal E 1 G antiserum (FKA‐224E; Cosmo Bio Co., Ltd., Tokyo, Japan) was raised in rabbits against estrone‐3‐glucuronide‐BSA and subsequently cross‐reacted with 100% estrone, 170% E 1 G, 25% estrone‐3‐sulfate, 1% estradiol, 0.1% estriol, 1.2% estradiol‐3‐glucuronide, 0.1% estradiol‐3‐sulfate, 0.05% testosterone, 0.07% 4‐androstenediol, 0% progesterone, 0% cortisol, and 0% 17‐α‐OH‐progesterone.…”

“…Urine creatinine concentrations were measured using the same methods and reagents described by Hama et al . The concentrations of steroid hormones and protease were calculated as ng or μg/mg of creatinine (Crmg).…”

“…Urine supernatant was used for the assays. Urine samples were Urine concentrations of E 1 G and PdG were assayed using an enzyme immunoassay using the same methods and reagents as described by Hama et al 22,23 The polyclonal E 1 G antiserum (FKA-224E;…”

Rapid Comparison of Metabolites in Humans and Rats of Different Sexes Using Untargeted Ultraperformance Liquid Chromatography Coupled to Time-of-Flight Mass Spectrometry and an in-house Software Platform

“…17 Marker a4 showed higher signal intensities in women than in men and was putatively annotated as pregnanediol-3-glucuronide or 3a,20a-dihydroxy-5b-pregnane 3-glucuronide, natural human metabolites of pregnanediol or 3a,20a-dihydroxy-5b-pregnane, respectively, and generated in the liver by UDP glucuronyltransferase (HMDB description, http://www.hmdb.ca/ metabolites/HMDB10318 and http://www.hmdb.ca/metabolites/HMDB10352), or isomers. Pregnanediol-3-glucuronide is a metabolite of progesterone present in large amounts in the urine of pregnant animals 18 and has long been proposed in the scientific literature to help identify the ovulatory period. 19 Pregnanediol, pregnanedione and pregnanolone are products of the pathway along which progesterone is primarily metabolized.…”

Functional annotation from the genome sequence of the giant panda

“…Previous studies have used genetic methods to extract phylogenetic and heterogeneous information on the giant panda (Pagés et al, 2008;Hama et al, 2009;Hao et al, 2009). Research has shown that the karyotype of the giant panda is similar to that of the bear (Nash et al, 1998), suggesting that the bear and the giant panda have similar genome sizes.…”