scholarly journals 246.The effect of testosterone and season on prodynorphin mRNA expression in the preoptic area hypothalamus of the ram

2004 ◽  
Vol 16 (9) ◽  
pp. 246
Author(s):  
C. J. Scott ◽  
I. J. Clarke ◽  
A. J. Tilbrook
Keyword(s):  
Mrna Expression ◽  
Breeding Season ◽  
Preoptic Area ◽  
Testosterone Propionate ◽  
In Situ Hybridisation ◽  
Pulse Amplitude ◽  
Plasma Testosterone ◽  
Mrna Levels ◽  
Bed Nucleus ◽  
Prodynorphin Mrna

Progesterone stimulates prodynorphin mRNA expression in the hypothalamus of the ewe (1), but whether testosterone regulates dynorphin gene expression in the ram is unknown. In a previous study (2), we showed that both testosterone and season influence mRNA expression of another opioid, enkephalin, in the preoptic area and hypothalamus of rams. Using tissue from the same study, we tested the hypothesis that testosterone and/or season modulate prodynorphin mRNA expression in specific areas of the hypothalamus in the ram. Adult Romney Marsh rams were castrated either during the 'breeding' season or 'non-breeding' season and 1 week later received intramuscular injections of either peanut oil (vehicle) or testosterone propionate (8 mg/12 h for 7 days) (5/group). Blood samples taken every 10 min for 12 h were assayed for plasma LH and testosterone. Prodynorphin mRNA expression was quantified in hypothalamic sections by in situ hybridisation using a 35S-labelled riboprobe and computer-aided image analysis. Plasma testosterone levels were higher in testosterone propionate-treated than oil-treated sheep. Mean plasma LH concentrations were reduced and the interpulse interval for LH pulses was greater in testosterone propionate-treated wethers compared to oil-treated wethers, with no change in LH pulse amplitude. Testosterone propionate treatment increased prodynorphin mRNA expression in the supraoptic nucleus and the bed nucleus of the stria terminalis, but only during the breeding season. Proenkephalin mRNA expression was also higher in the 'breeding' season than in the 'non-breeding' season in the caudal preoptic area and paraventricular nucleus. No differences were observed between treatments in five other regions of the hypothalamus. We conclude that testosterone and season regulate preproenkephalin mRNA levels in the preoptic area/hypothalamus in the ram in a region-specific manner. (1) Foradori et al. (2002) Proc. Soc. Neurosci. A572.9. (2) Scott et al. (2003) Biol. Reprod. 69, 2015–2021.

The role of neuropeptide Y (NPY) in the control of LH secretion in the ewe with respect to season, NPY receptor subtype and the site of action in the hypothalamus

1995 ◽  
Vol 147 (3) ◽  
pp. 565-579 ◽  
Author(s):  
M L Barker-Gibb ◽  
C J Scott ◽  
J H Boublik ◽  
I J Clarke
Keyword(s):  
Neuropeptide Y ◽  
Breeding Season ◽  
Preoptic Area ◽  
Pulse Amplitude ◽  
Pulse Frequency ◽  
Suppressive Effect ◽  
Plasma Prolactin ◽  
Post Injection ◽  
Lh Secretion ◽  
Saline Vehicle

Abstract Neuropeptide Y1–36 (NPY1–36) acts through Y1 and Y2 receptors while the C-terminal NPY fragments NPY18–36 and N-acetyl[Leu28,31]pNPY24–36 act only through the Y2 receptor. We have investigated the effects of intracerebroventricular (i.c.v.) administration of NPY1–36, NPY18–36 and N-acetyl[leu28,31]pNPY24–36 on LH secretion in the ovariectomised (OVX) ewe. These peptides were administered into a lateral ventricle (LV) or the third ventricle (3V) of OVX ewes during the non-breeding and breeding seasons. Microinjections of NPY were also made into the preoptic area (POA) during both seasons to investigate the effects of NPY at the level of the GnRH cell bodies. Tamed sheep were fitted with 19 gauge guide tubes into the LV, 3V or the septo-preoptic area (POA). Jugular venous blood samples were taken every 10 min for 3 h. Sheep were then given NPY1–36 (10 μg), NPY18–36 (100 μg) or saline vehicle into the LV; N-acetyl[Leu28,31]pNPY24–36 (100 μg), NPY1–36 (10 μg or 100 μg), NPY18–36 (10 μg or 100 μg) or saline vehicle into the 3V, or NPY1–36 (1 μg, 5 μg, 10 μg) into the POA. Blood sampling continued for a further 3 h. LH was measured in plasma by radioimmunoassay. LV or 3V injection of 10 μg NPY1–36 caused a small but significant (P<0·025) increase in the interval from the last pre-injection pulse of LH to the first post-injection LH pulse during the breeding season. Other LH pulse parameters were not significantly affected. NPY18–36 did not produce any significant change in LH pulsatility when injected into the LV, and neither peptide had any effect on plasma prolactin or GH levels. There was a significant (P<0·01) reduction in LH pulse frequency after 3V injection of 10 μg and 100 μg NPY and 100 μg NPY18–36. Pulse amplitude was reduced by 3V administration of the Y2 agonist, N-acetyl[Leu28–31]pNPY24–36 and 100 μ NPY18–36. When the amplitude of the first post-injection LH pulse was analysed, 10 μg NPY also had a significant (P<0·05) suppressive effect. During the non-breeding season, 100 μg NPY1–36 (but not 10 μg) decreased (P<0·01) LH pulse frequency. LH pulse amplitude was significantly (P<0·01) decreased by 100 μg NPY18–36. Doses of 10 μg NPY1–36 and 100 μg NPY18–36 had greater inhibitory effects on pulse frequency during the breeding season but the suppressive effect of 100 μg NPY was similar between seasons. Microinjections of NPY into the POA decreased (P<0·01) average plasma LH levels during the non-breeding season at a dose of 10 μg but did not significantly affect pulse frequency or amplitude. We conclude that a substantial component of the inhibitory action of NPY on LH secretion in the absence of steroids is mediated by the Y2 receptor. This inhibition is probably exerted by way of a presynaptic action on GnRH terminals in the median eminence as NPY does not modulate the frequency or amplitude of LH pulses at the level of the GnRH cell bodies in the POA. Journal of Endocrinology (1995) 147, 565–579

scholarly journals Physiological consequences of social descent: studies in Astatotilapia burtoni

2006 ◽  
Vol 190 (1) ◽  
pp. 183-190 ◽  
Author(s):  
Victoria N Parikh ◽  
Tricia Clement ◽  
Russell D Fernald
Keyword(s):  
Mrna Expression ◽  
Reproductive Capacity ◽  
Plasma Testosterone ◽  
Complete Regression ◽  
Mrna Levels ◽  
Short Term ◽  
Reproductive Regulation ◽  
Astatotilapia Burtoni ◽  
The Brain ◽  
Social Suppression

In many species, social interactions regulate reproductive capacity, although the exact mechanisms of such regulation are unclear. Since social stress is often related to reproductive regulation, we measured the physiological signatures of change in reproductive state as they relate to short-term stress and the stress hormone cortisol. We used an African cichlid fish, Astatotilapia burtoni, with two distinct, reversible male phenotypes: dominant (territorial, T) males that are larger, more brightly colored, more aggressive, and reproductively competent and non-dominant males (non-territorial, NT) that are smaller, camouflage colored, and have regressed gonads. Male status, and hence reproductive competence, depends on social experience in this system. Specifically, if a T male is placed among larger male fish, it quickly becomes NT in behavior and coloration, but complete regression of its reproductive axis takes ca. 3 weeks (White et al. 2002). Reproduction in all vertebrates is controlled by the hypothalamic–pituitary–gonadal axis in which the key signaling molecule from the brain to the pituitary is GnRH1. Here, we subjected T males to territory loss, a social manipulation which results in status descent. We measured the effects of this status change in levels of circulating cortisol and testosterone as well as mRNA levels of GnRH1 and GnRH receptor-1 (GnRH-R1) in the brain and pituitary, respectively. Following short-term social suppression (4 h), no change was observed in plasma cortisol level, GnRH1 mRNA expression, GnRH-R1 mRNA expression, or plasma testosterone level. However, following a somewhat longer social suppression (24 h), cortisol and GnRH1 mRNA levels were significantly increased, and testosterone levels were significantly decreased. These results suggest that in the short run, deposed T males essentially mount a neural ‘defense’ against loss of status.

scholarly journals Assessment of Testicular Lhcgr mRNA Expression Correlated with Testis and Seminal Vesicle Activities in the Libyan jird (Meriones libycus, Rodentia: Muridae) during Breeding Season Compared with Nonbreeding Season

Animals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 320
Author(s):  
Radia Boufermes ◽  
Mansouria Belhocine ◽  
Zaina Amirat ◽  
Farida Khammar
Keyword(s):  
Mrna Expression ◽  
Breeding Season ◽  
Luteinizing Hormone Receptor ◽  
Plasma Testosterone ◽  
Nonbreeding Season ◽  
Desert Rodent ◽  
Histological Method ◽  
Seasonal Breeder ◽  
Polymerase Chain ◽  
Breeding Seasons

The Libyan jird (Meriones libycus, 1823) is a wild desert rodent that is a seasonal breeder species adapted to breed when the environmental conditions can satisfy the energy and hydrous requirements of pregnant and nursing females to ensure that births occur at the most favorable time of the year. We assessed gene expression of testicular luteinizing hormone receptor (Lhcgr) correlated to testis activity. The expression of Lhcgr was evaluated using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR and the testis activity by a histological method in adult male Libyan jirds during the nonbreeding and breeding seasons. Our results showed that Lhcgr mRNA expression increased in autumn during the nonbreeding season and decreased in spring during the breeding season. This expression varied in contrast to testicular structure or function and plasma testosterone levels. These results help to elucidate this desert rodent’s seasonal sexual activity, which is correlated with central regulation.

scholarly journals Seasonal changes of androgen receptor, estrogen receptors and aromatase expression in the medial preoptic area of the wild male ground squirrels (Citellus dauricus Brandt)

2016 ◽  
Vol 60 (2) ◽  
Author(s):  
F. Zhang ◽  
J. Wang ◽  
Y. Jiao ◽  
L. Zhang ◽  
H. Zhang ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Estrogen Receptors ◽  
Breeding Season ◽  
Ground Squirrel ◽  
Preoptic Area ◽  
Medial Preoptic Area ◽  
Ground Squirrels ◽  
Mrna Levels ◽  
Wild Male ◽  
Significant Difference

<p>The wild ground squirrel is a typical seasonal breeder. In this study, using RT-PCR, western blot and immunohistochemistry, we investigated the mRNA and protein expressions of androgen receptor (AR), estrogen receptors α and β (ERα and ERβ) and aromatase cytochrome P450 (P450arom) in the medial preoptic area (MPOA) of hypothalamus of the wild male ground squirrel during the breeding season (April), the non-breeding season (June) and pre-hibernation (September). AR, ERα, ERβ and P450arom protein/mRNA were present in the MPOA of all seasons detected. The immunostaining of AR and ERα showed no significant changes in different periods, whereas ERβ and P450arom had higher immunoreactivities during the breeding season and pre-hibernation when compared to those of the non-breeding season. Consistently, both the protein and mRNA levels of P450arom and ERβ were higher in the MPOA of pre-hibernation and the breeding season than in the non-breeding season, whereas no significant difference amongst the three periods was observed for AR and ERα levels. These findings suggested that the MPOA of hypothalamus may be a direct target of androgen and estrogen. Androgen may play important regulatory roles through its receptor and/or the aromatized estrogen in the MPOA of hypothalamus of the wild male ground squirrels.</p>

Nuclear-specific accumulation of telomerase reverse transcriptase (TERT) mRNA in TERT promoter mutated follicular thyroid tumours visualised by in situ hybridisation: a possible clinical screening tool?

2021 ◽  
pp. jclinpath-2021-207631
Author(s):  
L Samuel Hellgren ◽  
Ann Olsson ◽  
Ann Kaufeldt ◽  
Johan O Paulsson ◽  
Martin Hysek ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reverse Transcriptase ◽  
Mrna Expression ◽  
In Situ Hybridisation ◽  
Telomerase Reverse Transcriptase ◽  
Mrna Levels ◽  
Specific Accumulation ◽  
Or Gene ◽  
Promoter Mutations

AimsUpregulation of the telomerase reverse transcriptase (TERT) gene is a frequent finding in follicular thyroid carcinomas (FTCs) with metastatic features. The augmented expression is usually caused by TERT promoter mutations. As TERT protein immunohistochemistry might not correlate to TERT mRNA levels in follicular thyroid tumours, we therefore sought to determine if visualisation of TERT mRNA through in situ hybridisation could highlight high-risk cases.MethodsWe collected formalin-fixated paraffin-embedded tissues from 26 follicular thyroid tumours; 7 FTCs, 2 follicular thyroid tumours of uncertain malignant potential (FT-UMPs) and a single Hürthle cell carcinoma with established TERT promoter mutations and gene expression, as well as 16 FTCs with no TERT gene aberrancy or gene expression, and assessed them using RNA Scope in situ hybridisation (ISH) and TERT probes targeting the two main TERT transcripts (TERT1 and TERT2).ResultsTERT 1 and/or 2 mRNA was found by ISH in 8/10 cases with established promoter mutations and mRNA expression, whereas all 16 cases without TERT gene aberrancies or gene expression were negative (Fisher’s exact p<0.001). Strikingly, TERT mRNA was visualised in the nuclear compartment only, thereby corroborating earlier studies suggesting a non-conventional role for TERT in tumour biology. Moreover, TERT mRNA expression was scattered across the tissue sections and only found in a few percentages of tumour nuclei.ConclusionsTERT mRNA seems to be focally expressed and localised exclusively to the nucleus in TERT promoter mutated follicular thyroid tumours, possibly reflecting a true biological and unorthodox phenomenon worthy of further investigations.

scholarly journals Postnatal Smoke Exposure Further Increases the Hepatic Nicotine Metabolism in Prenatally Smoke Exposed Male Offspring and Is Linked with Aberrant Cyp2a5 Methylation

2020 ◽  
Vol 22 (1) ◽  
pp. 164
Author(s):  
Khosbayar Lkhagvadorj ◽  
Zhijun Zeng ◽  
Karolin F. Meyer ◽  
Laura P. Verweij ◽  
Wierd Kooistra ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Mrna Expression ◽  
Nicotine Dependence ◽  
Susceptibility Gene ◽  
Smoke Exposure ◽  
Adverse Outcomes ◽  
Male Offspring ◽  
Adult Offspring ◽  
Mrna Levels ◽  
Nicotine Metabolism

Prenatal smoke exposure (PreSE) is a risk factor for nicotine dependence, which is further enhanced by postnatal smoke exposure (PostSE). One susceptibility gene to nicotine dependence is Cytochrome P450 (CYP) 2A6, an enzyme responsible for the conversion of nicotine to cotinine in the liver. Higher CYP2A6 activity is associated with nicotine dependence and could be regulated through DNA methylation. In this study we investigated whether PostSE further impaired PreSE-induced effects on nicotine metabolism, along with Cyp2a5, orthologue of CYP2A6, mRNA expression and DNA methylation. Using a mouse model where prenatally smoke-exposed adult offspring were exposed to cigarette smoke for 3 months, enzyme activity, mRNA levels, and promoter methylation of hepatic Cyp2a5 were evaluated. We found that in male offspring, PostSE increased PreSE-induced cotinine levels and Cyp2a5 mRNA expression. In addition, both PostSE and PreSE changed Cyp2a5 DNA methylation in male groups. PreSE however decreased cotinine levels whereas it had no effect on Cyp2a5 mRNA expression or methylation. These adverse outcomes of PreSE and PostSE were most prominent in males. When considered in the context of the human health aspects, the combined effect of prenatal and adolescent smoke exposure could lead to an accelerated risk for nicotine dependence later in life.

scholarly journals The usefulness of competitive PCR: airway gene expression of IL-5, IL-4, IL-4δ2, IL-2, and IFNγ in asthma

Thorax ◽  
2001 ◽  
Vol 56 (7) ◽  
pp. 541-548
Author(s):  
E M Glare ◽  
M Divjak ◽  
M J Bailey ◽  
E H Walters
Keyword(s):  
Gene Expression ◽  
Inhaled Corticosteroids ◽  
In Situ Hybridisation ◽  
Housekeeping Gene ◽  
Cross Sectional Study ◽  
Mrna Levels ◽  
Competitive Pcr ◽  
Cross Sectional ◽  
Steroid Use ◽  
Biopsy Specimens

BACKGROUNDAsthma has been described as an eosinophilic bronchitis driven by interleukin(IL)-4 and IL-5. The quantification of cytokine mRNA levels in airway samples has been confounded by housekeeping gene expression which differs between and within asthmatics and controls.METHODSThe usefulness of competitive reverse transcriptase-polymerase chain reaction (RT-PCR) that is independent of housekeeping gene expression for quantitating the mRNA for interferon (IFN)γ, IL-2, IL-5, IL-4 and its receptor antagonist encoding splicing variant IL-4δ2 was determined in a cross sectional study of 45 normal control subjects and 111 with asthma.RESULTSAtopic controls and atopic asthmatic subjects expressed more IL-5 than non-atopic controls (p<0.02) in bronchoalveolar lavage (BAL) cells, but not in biopsy specimens. IL-5 mRNA expression in BAL cells from asthmatic subjects using inhaled corticosteroids (ICS) was significantly lower than those not receiving ICS (p=0.04). IL-2 mRNA levels differed with steroid use in biopsy specimens but not in BAL cells. IFNγ, IL-4, and IL-4δ2 mRNA levels did not differ between any groups and were not affected by steroid use. IL-4 and IL-4δ2 mRNA levels were positively correlated (p<0.0001), suggesting coordinated transcription.CONCLUSIONSWhile the signal differentiation of competitive PCR in asthma may rival that of in situ hybridisation and immunohistochemistry, the method is expensive and wasteful of material.

scholarly journals Reprogramming mRNA Expression in Response to Defect in RNA Polymerase III Assembly in the Yeast Saccharomyces cerevisiae

2021 ◽  
Vol 22 (14) ◽  
pp. 7298
Author(s):  
Izabela Rudzińska ◽  
Małgorzata Cieśla ◽  
Tomasz W. Turowski ◽  
Alicja Armatowska ◽  
Ewa Leśniewska ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Ribosome Biogenesis ◽  
Rna Polymerase Iii ◽  
Mrna Levels ◽  
Rna Seq ◽  
Yeast Saccharomyces Cerevisiae ◽  
Protein Coding ◽  
General Transcription Factor ◽  
Pol I ◽  
Pol Iii

The coordinated transcription of the genome is the fundamental mechanism in molecular biology. Transcription in eukaryotes is carried out by three main RNA polymerases: Pol I, II, and III. One basic problem is how a decrease in tRNA levels, by downregulating Pol III efficiency, influences the expression pattern of protein-coding genes. The purpose of this study was to determine the mRNA levels in the yeast mutant rpc128-1007 and its overdose suppressors, RBS1 and PRT1. The rpc128-1007 mutant prevents assembly of the Pol III complex and functionally mimics similar mutations in human Pol III, which cause hypomyelinating leukodystrophies. We applied RNAseq followed by the hierarchical clustering of our complete RNA-seq transcriptome and functional analysis of genes from the clusters. mRNA upregulation in rpc128-1007 cells was generally stronger than downregulation. The observed induction of mRNA expression was mostly indirect and resulted from the derepression of general transcription factor Gcn4, differently modulated by suppressor genes. rpc128-1007 mutation, regardless of the presence of suppressors, also resulted in a weak increase in the expression of ribosome biogenesis genes. mRNA genes that were downregulated by the reduction of Pol III assembly comprise the proteasome complex. In summary, our results provide the regulatory links affected by Pol III assembly that contribute differently to cellular fitness.

scholarly journals Ex vivo mRNA expression of toll-like receptors during latent tuberculosis infection

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Birhan Alemnew ◽  
Soren T. Hoff ◽  
Tamrat Abebe ◽  
Markos Abebe ◽  
Abraham Aseffa ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Expression Patterns ◽  
Fold Change ◽  
Mrna Levels ◽  
Healthy Children ◽  
Toll Like Receptors ◽  
Relative Expression ◽  
Latently Infected ◽  
Latent Tb ◽  
Latent Tb Infection

Abstract Background Understanding immune mechanisms, particularly the role of innate immune markers during latent TB infection remains elusive. The main objective of this study was to evaluate mRNA gene expression patterns of toll-like receptors (TLRs) as correlates of immunity during latent TB infection and further infer their roles as potential diagnostic biomarkers. Methods Messenger RNA (mRNA) levels were analysed in a total of 64 samples collected from apparently healthy children and adolescents latently infected with tuberculosis (n = 32) or non-infected (n = 32). Relative expression in peripheral blood of selected genes encoding TLRs (TLR-1, TLR-2, TLR-4, TLR-6 and TLR-9) was determined with a quantitative real-time polymerase chain reaction (qRT-PCR) using specific primers and florescent labelled probes and a comparative threshold cycle method to define fold change. Data were analysed using Graph-Pad Prism 7.01 for Windows and a p-value less than 0.05 was considered statistically significant. Results An increased mean fold change in the relative expression of TLR-2 and TLR-6 mRNA was observed in LTBI groups relative to non-LTBI groups (p < 0.05), whereas a slight fold decrease was observed for TLR-1 gene. Conclusions An increased mRNA expression of TLR-2 and TLR-6 was observed in latently infected individuals relative to those non-infected, possibly indicating the roles these biomarkers play in sustenance of the steady state interaction between the dormant TB bacilli and host immunity.

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