Steroid profiling in male wobbler mouse, a model of Amyotrophic Lateral Sclerosis

The Wobbler mouse is an animal model for human motoneuron diseases, especially amyotrophic lateral sclerosis (ALS), used in the investigation of both pathology and therapeutic treatment. ALS is a fatal neurodegenerative disease, characterized by the selective and progressive death of motoneurons, leading to progressive paralysis. Previous limited studies have reported steroidal hormone dysregulation in Wobbler mouse and in ALS patients, suggesting endocrine dysfunctions which may be involved in the pathogenesis of the disease. In this study, we established a steroid profiling in brain, spinal cord, plasma, adrenal glands, and testes in 2-month-old male Wobbler mice and their littermates by gas chromatography coupled to mass spectrometry. Our results show in Wobbler mice the following: 1) a marked up-regulation of corticosterone levels in adrenal glands, plasma, spinal cord regions (cervical, thoracic, lumbar) and brain; 2) a strong decrease in T levels in the testis, plasma, spinal cord, and brain; and 3) increased levels of progesterone and especially of its reduced metabolites 5α-dihydroprogesterone, allopregnanolone, and 20α-dihydroprogesterone in the brain, spinal cord, and adrenal glands. Furthermore, Wobbler mice showed a hypothalamic-pituitary-gonadal hypoactivity. Interestingly, plasma concentrations of corticosterone and T correlate well with their respective levels in cervical spinal cord in both control and Wobbler mice. T down-regulation is probably the consequence of adrenal hyperactivity, and the up-regulation of progesterone and its reduced metabolites may correspond to an endogenous protective mechanism in response to motoneuron degeneration. Our findings suggest that increased levels of corticosterone and decreased levels of T in plasma could be a signature of motoneuron degeneration.Fil: Gonzalez Deniselle, Maria Claudia. Universidad de Buenos Aires. Facultad de Medicina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Liere, Philippe. Inserm; Francia. Université Paris Saclay; FranciaFil: Pianos, Antoine. Inserm; Francia. Université Paris Saclay; FranciaFil: Meyer, Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Aprahamian, Fanny. Inserm; Francia. Université Paris Saclay; FranciaFil: Cambourg, Annie. Inserm; Francia. Université Paris Saclay; FranciaFil: Di Giorgio, Noelia Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Schumacher, Michael. Inserm; Francia. Université Paris Saclay; Francia. Universite Paris Sud; FranciaFil: de Nicola, Alejandro Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Guennoun, Rachida. Université Paris Saclay; Francia. Inserm; Francia. Universite Paris Sud; Franci

Evolution of steroid concentrations in saliva from immature to pubertal gilts for the identification of biomarkers of gilts receptivity to boar effect

Estrus synchronization is necessary for management of gilt reproduction in pig farms. It is usually achieved by using synthetic progestagens, but there is increasing demand for non-hormonal alternative tools with the prospect of sustainability of livestock production. Moreover, in organic farms, synthetic hormones are not allowed. Before reaching puberty, gilts exhibit a “waiting period” during which external stimulations, such as boar exposure, could trigger and synchronize the first ovulation. However, practical non-invasive tools for detection of the “waiting period” in pig farms are lacking. During this period, estrone levels in urine are high, but urine sampling is difficult in group-housed females. Our objective was to identify among steroids potential biomarkers of this “waiting period” through saliva monitoring from immature to pubertal gilts using gas chromatography coupled to tandem mass spectrometry. Starting between 144 and 147 days of age, six Large White gilts were submitted to ultrasound puberty diagnosis 3 times a week until first ovulation. Urine and saliva samples were collected to analyze weekly estrone and steroidome respectively, until puberty. Urinary estrone concentration significantly increased 2 weeks before first ovulation occurring between 182 and 192 days of age. The period with increasing estrone levels was considered as the “waiting period”. Steroidome analysis allowed identifying and quantifying 28 steroids in 500 µl of gilts saliva. Significant decrease of dehydroepiandrosterone and significant increase of 5α-dihydroprogesterone and 17β-estradiol were detected 2 weeks before puberty, suggesting that these steroids could be potential biomarkers of the “waiting period”. These results show that painless sampling of saliva could be a non-invasive welfare-friendly tool for the identification of the physiological hormonal status of the gilts and possibly the optimal time for application of the boar effect, a solution to synchronize puberty without exogenous hormones

Steroidome And Metabolome Analysis In Saliva From Immature To Pubertal Gilts To Identify Potential Biomarkers Of Receptivity To Boar Effect

Our objective was to develop alternatives to hormones for estrus synchronization in gilts. Gilts exhibit a pre-puberty period with high urinary estrone concentration during which boar exposure could induce the first ovulation. We searched for salivary biomarkers of this period. Urine and saliva were collected on six 140-day-old gilts until puberty for estrone assay, metabolome and steroidome analysis. We identified 23 metabolites and 28 steroids in saliva. The concentration of 8 of them showed significant variations at the pre-puberty period, they were candidate biomarkers. Saliva was collected from 30 gilts exposed to a boar and subjected to estrus detection from 150 to 175 days of age. Metabolome and steroidome analyses allowed the identification of 33 metabolites and 29 steroids in saliva. Their concentrations were not significantly different between receptive and non-receptive gilts. Thus, we could not identify salivary biomarkers of the period of receptivity to the boar effect

Abnormal steroidogenesis and aromatase activity in preeclampsia.

International audienceIntroduction :Estrogens and progesterone play critical roles in angiogenesis and vasodilation. Moreover, placental aromatase deficiency is detected in women with preeclampsia (PE) at delivery. We hypothesized that abnormal steroidogenesis occurs much earlier than typical PE diagnosis. Thus, we investigated whether the circulating steroid profile was already disturbed at 24–29 weeks of gestation in women with subsequent PE, and compared the profile with that of women with “placental” small gestational age (SGA) without PE.Methods :We selected nulliparous women (n = 90) from the MOMA trial, including women with PE (n = 25), SGA (n = 25), and controls (NP; n = 40), for plasma steroid profiling by gas chromatography/mass spectrometry and to measure placental growth factor and soluble fms-like tyrosine kinase-1. Placental aromatase expression was evaluated in a new set of women.Results :Compared with that of controls, the women with PE had a significantly lower estrone/androstenedione ratio, and exhibited a decreasing trend for estradiol and estrone levels. Lower estriol levels were observed in the SGA group compared to the NP group. Compared with that of controls, the women with PE and SGA had significantly higher levels of 20α-dihydroprogesterone (20α-DHP) and 20α-DHP/progesterone ratios. Pregnenolone sulfate levels were lower in the PE group than in the NP and SGA groups. Decreased expression of aromatase was observed in the PE group compared to the control group.Discussion :Preeclampsia appears to be characterized by specific steroidogenesis dysregulation long before PE diagnosis, highlighting potential new biomarkers of PE.

Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key

Pregnenolone (PREG) and dehydroepiandrosterone (DHEA), and their respective sulfated forms PREGS and DHEAS, were among the first steroids to be identified in rodent brain. However, unreliable steroid isolation and solvolysis procedures resulted in errors, particularly in the case of brain steroid sulfates analyzed by radioimmunology or GC-MS of liberated free steroids. By using a solid-phase extraction recycling/elution procedure, allowing the strict separation of sulfated, free, and fatty acid esters of PREG and DHEA, PREGS and DHEAS, unlike free PREG, were not detected in rat and mouse brain and plasma. Conversely, considerable amounts of PREG and DHEA were released from unknown precursor(s) present in the lipoidal fraction, distinct from fatty acid ester conjugates. Chromatographic and mass spectrometric studies of the nature of the precursor(s) showed that autoxidation of brain cholesterol (CHOL) was responsible for the release of PREG and DHEA from the lipoidal fraction. When inappropriate protocols were used, CHOL was also the precursor of PREG and DHEA obtained from the fraction assumed to contain sulfated steroids. In contrast, free PREG was definitely confirmed as an endogenous steroid in rat brain. Our study shows that an early removal of CHOL from brain extracts coupled to well-validated extraction and fractionation procedures are prerequisites for reliable measurements of free and conjugated PREG and DHEA by GC-MS or other indirect methods