Glucocorticoid production in lymphoid organs: Acute effects of lipopolysaccharide in neonatal and adult mice

Endocrinology ◽  
2021 ◽  
Author(s):  
Melody Salehzadeh ◽  
Jordan E Hamden ◽  
Michael X Li ◽  
Hitasha Bajaj ◽  
Ruolan S Wu ◽  
...  
Keyword(s):  
Hpa Axis ◽  
Adrenal Glands ◽  
Lymphoid Organs ◽  
Acute Effects ◽  
Steroidogenic Enzymes ◽  
Steroidogenic Enzyme ◽  
Adult Mice ◽  
Liquid Chromatography Tandem Mass ◽  
Glucocorticoid Production ◽  
Local Expression

Abstract Glucocorticoids (GCs) are critical modulators of the immune system. The hypothalamic-pituitary-adrenal (HPA) axis regulates circulating GC levels and is stimulated by endotoxins. Lymphoid organs also produce GCs; however, it is not known how lymphoid GC levels are regulated in response to endotoxins. We assessed whether an acute challenge of lipopolysaccharide (LPS) increases lymphoid levels of GCs, steroidogenic enzymes expression, and components of the HPA axis (e.g., CRH) expression. We administered LPS (50µg/kg i.p.) or vehicle control to male and female C57BL/6J neonatal (post-natal day (PND) 5) and adult (PND90) mice and collected blood, bone marrow, thymus, and spleen 4 hr later. We measured progesterone, 11-deoxycorticosterone (DOC), corticosterone, and 11-dehydrocorticosterone (DHC) via liquid chromatography tandem mass spectrometry (LC-MS/MS). We measured gene expression of key steroidogenic enzymes (Cyp11b1, Hsd11b1, and Hsd11b2) and HPA axis components (Crh, Crhr1, Pomc, and Mc2r) via qPCR. At PND5, LPS induced greater increases in steroid levels in lymphoid organs than in blood. In contrast, at PND90, LPS induced greater increases in steroid levels in blood than in lymphoid organs. Steroidogenic enzyme transcripts were present in all lymphoid organs, and LPS altered steroidogenic enzyme expression predominately in the spleen. Lastly, we detected transcripts of key HPA axis components in all lymphoid organs, and there was an effect of LPS in the spleen. Taken together, these data suggest that LPS regulates GC production by lymphoid organs, similar to its effects on the adrenal glands, and the effects of LPS might be mediated by local expression of CRH and ACTH.

scholarly journals OR19-04 Glucocorticoid Production in the Nervous and Immune Systems: Evidence for a Local HPA Axis Homolog

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Melody Salehzadeh ◽  
Jordan Edward Hamden ◽  
Michael X Li ◽  
Hitasha Bajaj ◽  
Cathy Ma ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Hpa Axis ◽  
Anterior Pituitary ◽  
Adrenal Glands ◽  
Brain Regions ◽  
Lymphoid Organs ◽  
Thymocyte Development ◽  
Immune Systems ◽  
Immune System Development ◽  
Glucocorticoid Production

Abstract Glucocorticoid Production in the Nervous and Immune Systems: Evidence for a Local HPA Axis Homolog The hypothalamic-pituitary-adrenal (HPA) axis is a critical stress response system in vertebrates. The hypothalamus secretes corticotropin-releasing hormone (CRH), which binds its receptor (CRH-R1) in the anterior pituitary. The anterior pituitary then secretes adrenocorticotropic hormone (ACTH), which binds its receptor (MC2R) in the adrenal glands and stimulates secretion of glucocorticoids into the bloodstream. Glucocorticoids are critical modulators of neural and immune system development. During early development (postnatal day (PND) 2 to 12), mice show decreased adrenal glucocorticoid secretion at baseline and in response to stressors, termed the stress hyporesponsive period (SHRP) (1). Traditionally, glucocorticoids have been thought to be synthesized only in the adrenal glands. However, recent evidence demonstrates that glucocorticoids are also produced in extra-adrenal tissues, such as the brain and lymphoid organs (2). This may be of particular importance during the SHRP, as local production allows glucocorticoid modulation of specific tissues and cells, without general effects throughout the organism. Importantly, the factors that regulate local glucocorticoid production remain unknown. To study the regulation of local glucocorticoid production, we examined whether mediators of the HPA axis are locally expressed at baseline and in response to an immune stressor. We assessed systemic and local glucocorticoid levels in neonatal (PND5) C57BL/6J mice 4hr after an immune challenge with lipopolysaccharide (50µg/kg i.p.) or vehicle control. We examined blood, microdissected brain regions (prefrontal cortex, hippocampus, hypothalamus), and lymphoid organs (thymus, spleen, bone marrow). A panel of 7 steroids was measured via liquid chromatography tandem mass spectrometry (LC-MS/MS). Gene expression of Crh, Crh-R1, Pomc, and Mc2r was quantified via qPCR. Preliminary data indicate that corticosterone was 2-fold higher in tissues than in blood after an immune stressor. The thymus expressed all genes of interest, supporting the existence of a local HPA axis “homolog” in the thymus. Brain, spleen and bone marrow expressed a subset of the genes of interest. These exciting data demonstrate that all the mediators of the HPA axis are locally expressed within the thymus, likely to regulate thymocyte development and reactivity. Greater understanding of local glucocorticoid production will provide crucial insight into neural and immune development and function. Reference: (1) Sapolsky et al., Brain Res Rev. 1986 11(1):65–76. (2) Taves et al., Endocrinology. 2015 156(2):511–522.

Isoflurane stress induces glucocorticoid production in mouse lymphoid organs

2021 ◽  
Author(s):  
Jordan E Hamden ◽  
Melody Salehzadeh ◽  
Katherine M. Gray ◽  
Brandon J Forys ◽  
Kiran K Soma
Keyword(s):  
Adrenal Glands ◽  
Vehicle Control ◽  
Postnatal Period ◽  
Lymphoid Organs ◽  
Systemic Steroid ◽  
Potential Mechanism ◽  
Response To Stress ◽  
Glucocorticoid Production ◽  
Acute Stressor ◽  
Insight Into

Glucocorticoids (GCs) are secreted by the adrenal glands and locally produced by lymphoid organs. Adrenal GC secretion at baseline and in response to stressors is greatly reduced during the stress hyporesponsive period (SHRP) in neonatal mice (postnatal day (PND) 2-12). It is unknown whether lymphoid GC production increases in response to stressors during the SHRP. Here, we administered an acute stressor (isoflurane anesthesia) to mice before, during, and after the SHRP and measured systemic and local GCs via mass spectrometry. We administered isoflurane, vehicle control (oxygen), or neither (baseline) at PND 1, 5, 9, or 13 and measured progesterone and 6 GCs in blood, bone marrow, thymus, and spleen. At PND1, blood and lymphoid GC levels were high and did not respond to stress. At PND5, blood GC levels were very low and increased slightly after stress, while lymphoid GC levels were also low but, increased greatly after stress. At PND9, blood and lymphoid GC levels were similar at baseline and increased similarly after stress. At PND13, blood GC levels were higher than lymphoid GC levels at baseline, and blood GC levels showed a greater response to stress. These data demonstrate the remarkable plasticity of GC physiology during the postnatal period, show that local steroid levels do not reflect systemic steroid levels, provide insight into the SHRP, and identify a potential mechanism by which early-life stressors can alter immunity in adulthood.

The acute effects of multiple doses of methamphetamine on locomotor activity and anxiety-like behavior in adolescent and adult mice

2021 ◽  
Vol 405 ◽  
pp. 113186
Author(s):  
Hayley A. Ortman ◽  
Mikayla L. Newby ◽  
Jonathan Acevedo ◽  
Jessica A. Siegel
Keyword(s):  
Locomotor Activity ◽  
Acute Effects ◽  
Adult Mice ◽  
Multiple Doses

Thymostimulin effects on lymphoid organs in Ames dwarf mice

1993 ◽  
Vol 128 (1) ◽  
pp. 74-80
Author(s):  
Maria A Villanua ◽  
Agnieszka Szary ◽  
Ana I Esquifino ◽  
Andrzej Bartke
Keyword(s):  
Immune Function ◽  
Lymphoid Organs ◽  
Plasma Prolactin ◽  
Nk Activity ◽  
Spleen Cells ◽  
Adult Mice ◽  
Ames Dwarf Mice ◽  
Ames Dwarf ◽  
Spleen Lymphocytes ◽  
Dwarf Mice

This work was undertaken to study the effects of thymostimulin (TP-1) on the immune function in Ames dwarf mice, and to relate these effects to PRL and/or GH deficiency in these animals. Male Ames dwarf mice implanted with pituitaries from normal mice under the kidney capsule, sham-operated dwarf mice and normal immature or adult mice were injected daily for five days with TP-1. In comparison to normal animals, sham-operated dwarf mice had markedly lower body, thymus and spleen weights, as well as a lower number of lymphocytes in the spleen and in the thymus and the natural killer (NK) activity of spleen lymphocytes. Ectopic pituitary transplants produced the expected enhancement of body weight gain and increased spleen and thymus weights, which reached the values found in normal (non-dwarf) animals. The numbers of lymphocytes in the spleen and thymus were significantly increased in pituitary-grafted dwarf mice, but the grafts did not modify the cytotoxic activity of NK spleen cells, or the number of peripheral white blood cells (PWBC). In sham-operated dwarf mice, TP-1 treatment did not modify the number of cells in the spleen and thymus, or the NK activity. In pituitary-grafted dwarf mice, treatment with TP-1 induced an increase in the number of spleen lymphocytes and in the NK activity of spleen cells without affecting the weight of lymphoid organs or the number of thymic cells. Plasma prolactin (PRL) and growth hormone (GH) levels of pituitary-grafted dwarf mice were not changed after TP-1 administration. Surprisingly, the NK activity of spleen lymphocytes in normal adult mice was greatly increased after TP-1 administration. These findings suggest that the thymic extract TP-1 can exert a major stimulatory influence on NK activity of spleen lymphocytes in adult mice, and potentiate some of the stimulatory effects of hormones secreted by ectopic pituitary transplants on the immune function of Ames dwarf mice. These effects are not mediated by modifications of the release of PRL or GH.

scholarly journals Analysis of mRNA expression for steroidogenic enzymes in the remaining adrenal cortices attached to adrenocortical adenomas.

2008 ◽  
Vol 158 (6) ◽  
pp. 867-878 ◽  
Author(s):  
Kazuto Shigematsu ◽  
Takehiro Nakagaki ◽  
Naohiro Yamaguchi ◽  
Kioko Kawai ◽  
Hideki Sakai ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Zona Fasciculata ◽  
Steroidogenic Enzymes ◽  
Steroidogenic Enzyme ◽  
Adrenocortical Adenomas ◽  
Aldosterone Production ◽  
Aldosterone Producing Adenoma ◽  
Cortical Nodules ◽  
Design And Methods

Design and methodsWe have recently demonstrated that the adrenal cortices attached to aldosterone-producing adenoma (APA) contained microscopic subcapsular micronodules suggestive of active aldosterone production. In this study, we used in situ hybridization to investigate the mRNA expression of steroidogenic enzymes in the adrenal cortices attached to cortisol-producing adenoma (CPA) and clinically silent adenoma (non-functioning adenoma; NFA), in addition to APA.ResultsMicroscopic subcapsular micronodules, which were several hundreds of micrometers in size and spheroid in shape, were observed in the cortices attached to CPA and NFA, as well as APA, at high frequency. Most of the cortical nodules in zona fasciculata to zona reticularis showed a suppressed steroidogenesis in the cortices attached to adenoma, but some expressed intensely all necessary steroidogenic enzyme mRNAs for cortisol synthesis.ConclusionsIt is thus necessary to keep in mind, on the occasion of subtotal adrenalectomy, that lesions with the potential to later develop into functional adrenocortical nodules may be present in other parts of the ipsilateral or contralateral adrenal cortices.

scholarly journals Luteinizing Hormone/Human Chorionic Gonadotropin-Mediated Activation of mTORC1 Signaling Is Required for Androgen Synthesis by Theca-Interstitial Cells

2012 ◽  
Vol 26 (10) ◽  
pp. 1732-1742 ◽  
Author(s):  
Murugesan Palaniappan ◽  
K. M. J. Menon
Keyword(s):  
Chorionic Gonadotropin ◽  
Regulatory Protein ◽  
Steroidogenic Acute Regulatory Protein ◽  
Steroidogenic Enzymes ◽  
Steroidogenic Enzyme ◽  
S6 Kinase ◽  
Ribosomal Protein S6 ◽  
Mtorc1 Signaling ◽  
Steroidogenic Acute Regulatory ◽  
I Cells

Abstract LH triggers the biosynthesis of androgens in the theca-interstitial (T-I) cells of ovary through the activation of a cAMP-dependent pathway. We have previously shown that LH/human chorionic gonadotropin (hCG) activates mammalian target of rapamycin complex 1 (mTORC1) signaling network, leading to cell proliferation. In the present study, we provide evidence that the LH/hCG-mediated activation of the mTORC1 signaling cascade is involved in the regulation of steroidogenic enzymes in androgen biosynthesis. Treatment with LH/hCG increased the expression of downstream targets of mTORC1, ribosomal protein S6 kinase 1, and eukaryotic initiation factor 4E as well as steroidogenic enzymes. LH/hCG-mediated stimulation of the steroidogenic enzyme mRNA was blocked by the mTORC1 inhibitor, rapamycin. This inhibitory effect was selective because rapamycin failed to block hCG-mediated increase in the expression of Star mRNA levels. Furthermore, pharmacological targeting of mTORC1 with rapamycin also blocked LH/hCG- or forskolin-induced expression of cAMP response element-binding protein (CREB) and steroidogenic enzymes (P450 side-chain cleavage enzyme, 3β-hydroxysteroid dehydrogenase type 1, and 17α-hydroxylase/17,20 lyase) but produced no effect on steroidogenic acute regulatory protein levels. These results were further confirmed by demonstrating that the knockdown of mTOR using small interfering RNA selectively abrogated the LH/hCG-induced increase in steroidogenic enzyme expression, without affecting steroidogenic acute regulatory protein expression. LH/hCG-stimulated androgen production was also blocked by rapamycin. Furthermore, the pharmacological inhibition of mTORC1 or ribosomal protein S6 kinase 1 signaling prevented the LH/hCG-induced phosphorylation of CREB. Chromatin immunoprecipitation assays revealed the association of CREB with the proximal promoter of the Cyp17a1 gene in response to hCG, and this association was reduced by rapamycin treatment. Taken together, our findings show for the first time that LH/hCG-mediated activation of androgen biosynthesis is regulated by the mTORC1 signaling pathway in T-I cells.

Expression of steroidogenic enzymes in the bovine placenta and fetal adrenal glands throughout gestation.

Endocrinology ◽  
1992 ◽  
Vol 130 (5) ◽  
pp. 2641-2650 ◽  
Author(s):  
A J Conley ◽  
J R Head ◽  
D T Stirling ◽  
J I Mason
Keyword(s):  
Adrenal Glands ◽  
Steroidogenic Enzymes ◽  
Bovine Placenta

scholarly journals Ectopic Agouti protein overexpression increases stimulated corticosterone production without effect on adenylate cyclase activity in mouse adrenal cells

2004 ◽  
pp. 613-618 ◽  
Author(s):  
AY Shevchenko ◽  
NM Bazhan ◽  
EN Makarova ◽  
TV Yakovleva ◽  
NR Karkaeva
Keyword(s):  
Adenylate Cyclase ◽  
Protein Overexpression ◽  
Dibutyryl Camp ◽  
Steroidogenic Enzymes ◽  
Steroidogenic Enzyme ◽  
Adrenal Cells ◽  
Agouti Protein ◽  
Corticosterone Release ◽  
Agouti Related Protein ◽  
Enzyme Reactivity

OBJECTIVE: The antagonism of Agouti protein (AP) and Agouti-related protein on melanocortin receptors suggests an inhibitory role in the regulation of steroidogenesis. However, we have previously demonstrated that ectopic AP overexpression increased restraint-induced corticosterone release and adrenal reactivity to ACTH in mice. A high steroidogenic response to ACTH may be a consequence of a stimulatory AP action on the adenylate cyclase (AC) and/or intracellular steroidogenic enzymes. The aim of the present study was to estimate the effect of ectopic AP overexpression on the activity of AC and steroidogenic intracellular enzymes. METHODS: ACTH and forskolin were used for AC stimulation, and dibutyryl cAMP and progesterone were used for stimulation of intracellular steroidogenic enzymes in isolated adrenal cells in male C57Bl/6J mice of two Agouti genotypes: A(y)/a (ectopic AP overexpression) and a/a (absence of AP in all tissues). RESULTS: ACTH and forskolin increased cAMP accumulation to the same extent in both A(y)/a and a/a mouse adrenal cells (P<0.001; ANOVA), but resulted in higher corticosterone production in A(y)/a mice (P<0.001 for ACTH and P<0.01 for forskolin; ANOVA). Dibutyryl cAMP- and progesterone-induced corticosterone production was higher in A(y)/a mice than in a/a mice (P<0.001 for dibutyryl cAMP and P<0.01 for progesterone; ANOVA). CONCLUSIONS: Ectopic AP overexpression increased stimulated corticosterone production and intracellular steroidogenic enzyme reactivity to cAMP without an effect on AC activity.

Effects of C19 steroids on adrenal steroidogenic enzyme activities and their mRNA levels in guinea-pig fasciculata-glomerulosa cells in primary culture

1992 ◽  
Vol 132 (2) ◽  
pp. 269-276 ◽  
Author(s):  
P. H. Provencher ◽  
Y. Tremblay ◽  
A. Bélanger
Keyword(s):  
Guinea Pig ◽  
Primary Culture ◽  
Oxygen Tension ◽  
Enzyme Activities ◽  
Inhibitory Effect ◽  
Mrna Levels ◽  
Hydroxylase Activity ◽  
Steroidogenic Enzymes ◽  
Steroidogenic Enzyme ◽  
Steroid Synthesis

ABSTRACT The present study examined the effects of steroids on steroidogenic enzyme activity in adrenal glands. Guinea-pig fasciculata-glomerulosa (FG) cells maintained in primary culture were exposed to steroids for 48 h. Although the treatment with androstenedione alone had no effect on 3β-hydroxysteroid dehydrogenase 4-ene-5-ene-isomerase (3β-HSD), 17-hydroxylase and 17,20-lyase activities, there was inhibition of 11-hydroxylase and 21-hydroxylase activities. When FG cells were exposed to 10 nmol ACTH/l for the last 24 h of incubation, ACTH alone had no effect on steroidogenic enzymes but, while combined with androstenedione, it further decreased 21-hydroxylase activity and stimulated 17-hydroxylase and 17,20-lyase activities. Cortisol, corticosterone, oestradiol and 11β-hydroxy androstenedione had no effect on steroidogenic enzyme activities while the inhibitory effect on 21-hydroxylase activity was only observed with androstenedione, testosterone and dihydrotestosterone. Addition of hydroxyflutamide, a pure antiandrogen, did not block the inhibitory effect of androstenedione on 21-hydroxylase and 11-hydroxylase activities. The reduction in oxygen tension from 19 to 2% which was aimed at examining the oxygen-mediated effects on steroidogenic enzymes, revealed that the reduction in 21-hydroxylase activity induced by androstenedione could not be prevented by low oxygen tension. An interaction of C19 steroids at the level of the enzymes is also suggested by our finding that androstenedione had no effect on basal and ACTH-stimulated steady-state 11-hydroxylase, 17-hydroxylase, 17,20-lyase and 21-hydroxylase mRNA levels. These results indicate that C19 steroids alter the adrenal steroidogenic enzyme activities in such a manner that C19 steroid synthesis is increased while glucocorticoid production is inhibited. The mechanism of action of C19 steroids does not involve gene expression for steroidogenic enzymes but probably a direct interaction with steroidogenic enzymes, namely 21-hydroxylase, 17-hydroxylase and 17,20-lyase. Our data suggest that C19 steroids may reduce the amount of 21-hydroxylase in the microsomal fraction which may have a major impact on the levels of microsomal P450 reductase available for 17-hydroxylase and 17,20-lyase activities. Journal of Endocrinology (1992) 132, 269–276

scholarly journals Androgenic potential of human fetal adrenals at the end of the first trimester

2017 ◽  
Vol 6 (6) ◽  
pp. 348-359 ◽  
Author(s):  
I Savchuk ◽  
M L Morvan ◽  
J P Antignac ◽  
K Gemzell-Danielsson ◽  
B Le Bizec ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Transcription Factors ◽  
Western Blotting ◽  
Adrenal Glands ◽  
First Trimester ◽  
Gas Chromatography Mass Spectrometry ◽  
Steroidogenic Enzymes ◽  
Complex Process ◽  
Low Expression

The onset of steroidogenesis in human fetal adrenal glands (HFA) during the first trimester is poorly investigated. An unresolved question is the capacity of the HFA to produce potent androgen DHT via conventional and/or the backdoor pathway(s) at the end of first trimester, when androgen-responsive organs are developed. Our aim was to explore steroidogenesis and the expression of steroidogenic enzymes and transcription factors in HFA at gestational weeks (GW) 9–12 with focus on their androgenic potential. Steroids in the HFA were analyzed by gas chromatography/mass spectrometry. The expression of steroidogenic enzymes and transcription factors in the HFA at GW9–12 was investigated by qPCR, automated Western blotting and immunohistochemistry. We demonstrated that during GW9–12 HFA produced steroids of the ∆5, ∆4 and the backdoor pathways of the biosynthesis of DHT, though the latter was limited to production of 17α-OH-dihydroprogesterone, androsterone and androstanedione without further conversion to DHT. The only androgens identified in the HFA were testosterone and androsterone, a precursor in the biosynthesis of DHT. We also observed higher levels of CYP17A1 but low expression of 3βHSD2 at GW11–12 in the HFA. Elevated levels of CYP17A1 were associated with an increased expression of SF-1 and GATA-6. Altogether, our data demonstrate that of those steroids analyzed, the only potent androgen directly produced by the HFA at GW9–12 was testosterone. The onset of steroidogenesis in the HFA is a complex process that is regulated by the coordinated action of related transcription factors.

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