The thyroid C cells of ovariectomized rats treated with estradiol

The structure and function of thyroid C cells were studied in ovariectomized (Ovx) adult female rats without and after chronic treatment with estradiol dipropionate (EDP). A peroxidase–antiperoxidase method was applied for localization of calcitonin (CT) in the C cells. Morphometric changes in their volume, nuclei, and relative volume density were evaluated in comparison with sham-operated control rats using a stereological method. The number of C cells was calculated. CT content in the sera was determined by radioimmunoassay. Ovariectomy (Ovx) led to a 21% increase in body weight (P<0.005), while treatment of Ovx rats with EDP decreased body weight by 25% (P<0.01). The immunoreactivity for CT in C cells of the Ovx rats was markedly increased. Significant decreases in the volume of C cells (by 13%; P<0.05) and serum CT (by 45%) were recorded, while the C cell number increased by 59% (P<0.05) in relation to the corresponding controls. The treatment of Ovx rats with EDP caused conspicuous degranulation of the C cells. The cellular volume was increased by 11% and serum CT by 36% in comparison with Ovx animals. At the same time a decrease in C cell number by 29% (P<0.05) was evident. It may be concluded that estradiol deficiency after Ovx reduced the synthesis and release of CT, while chronic treatment of these animals with EDP had a positive effect on the secretory activity of thyroid C cells.     

Effects of estradiol and calcium on gonadotrophic cells in middle-aged female rats

The effects of multiple treatment with estradiol dipropionate (EDP) or calcium glucoheptonate (Ca) or a combination of the two on gonadotrophic cells in the pituitary pars distalis of middle-aged female rats were examined. The animals were treated daily for two weeks with EDP (0.625mg i.p./kg body weight) or Ca (11.4mg/kg body weight) or EDP+Ca. Luteinising (LH) and follicle stimulating hormone (FSH)-producing cells were examined by immunohistochemistry using antisera to the specific (beta) -subunits of LH and FSH and a peroxidase–anti-peroxidase immunohistochemical procedure. Plasma levels of FSH and LH were measured by radio-immune assay. A stereological method for determining morphometric parameters in immunopositive FSH and LH cells was used. The number of gonadotrophs per unit area (mm²), their cellular volume and relative volume densities, as well as plasma levels of FSH and LH, were decreased in all treated females in comparison with the controls. The most significant decrease of these parameters was observed in EDP-treated animals. Such changes were also expressed in Ca-treated animals, but the alterations were less distinct. These results demonstrate that multiple EDP or Ca application to middle-aged female rats is able to inhibit, directly or indirectly, the morphofunctional state of gonadotrophic cells in the pituitary pars distalis.     

Voluntary exercise impairs initial delayed spatial alternation performance in estradiol treated ovariectomized middle-aged rats

Estrogens differentially modulate behavior in the adult female rodent. Voluntary exercise can also impact behavior, often reversing age associated decrements in memory processes. Our research group has published a series of papers reporting a deficit in the acquisition of an operant working memory task, delayed spatial alternation (DSA), following 17β-estradiol treatment to middle-aged ovariectomized (OVX) rats. The current study examined if voluntary exercise could attenuate the 17β-estradiol induced deficits on DSA performance. OVX 12-month old Long–Evans rats were implanted with a Silastic capsule containing 17β-estradiol (10% in cholesterol: low physiological range) or with a blank capsule. A subset of the 17β-estradiol and OVX untreated rats were given free access to a running wheel in their home cage. All rats were tested for 40 sessions on the DSA task. Surprisingly, we found running wheel access to impair initial acquisition of the DSA task in 17β-estradiol treated rats, an effect not seen in OVX untreated rats given running wheel access. This deficit was driven by an increase in perseverative responding on a lever no longer associated with reinforcement. We also report for the first time a 17β-estradiol induced impairment on the DSA task following a long intertrial delay (18-sec), an effect revealed following more extended testing than in our previous studies (15 additional sessions). Overall, running wheel access increased initial error rate on the DSA task in 17β-estradiol treated middle-aged OVX rats, and failed to prevent the 17β-estradiol induced deficits in performance of the operant DSA task in later testing sessions.     

The adrenal cortex in female rats after estradiol or calcium treatment

Administration of estradiol or calcium, or combined, represents the classical therapeutic approach in the treatment of some menopausal symptoms. We have studied the effects of estradiol dipropionate (EDP) and calcium glucoheptonate (Ca) on morphological and hormonal features of the adrenal gland in 14-month-old female Wistar rats. The animals were treated with EDP (0.625 mg/kg b.w.) or Ca (11.4 mg/kg b.w.) daily for two weeks, with control rats receiving vehicle alone by the same schedule. The cell volumes in the zona glomerulosa (ZG) and zona fasciculata (ZF) were 11.2% and 5.5% greater (P<0.05) and in the zona reticularis (ZR) 13.0% smaller (P<0.05) in the EDP group than in the control group. In the Ca group, cell volume in the ZG was increased by 5.6% (P<0.05), while cell volumes in the ZF and ZR were decreased by 26.0% and 14.7%, respectively (P<0.05), in comparison with control values. Serum aldosterone and corticosterone concentrations were higher in the EDP-treated (by 27.8% and 19.8%, respectively) and Ca-treated (by 80.0% and 24.1%, respectively) groups in comparison with the control group (P<0.05). These data suggest that EDP and Ca treatments have stimulatory effects on the ZG and ZF, but inhibitory effects on the ZR in middle-aged female rats.     

Immunocytological studies of C cells with anti-calcitonin or antisomatostatin immunoserums in rats treated with vitamin D, thyroxine or benzyl-thiouracil]

In rats treated with vitamin D3, thyroxin or BTU alone or with associations of vitamin D3 + BTU or vitamin D + thyroxin, immunocytochemical studies with an anti-human calcitonin serum show a stimulating effect of vitamin D and of thyroxin and an inhibiting effect of BTU on the C cells of the thyro?d. These results are in agreement with the existence of functional relations between follicular and C cells. A few calcitonin-containing cells are immunoreactive to an anti-somatostatin serum. All the C cells contain both calcitonin and somatostatin when they are grouped in a single big interfollicular cluster.     

Thyroid C cells in male and female rats with chronic renal failure

The kidneys are responsible for iodine and of thyroid hormone biodegradation. The aim of this study was the histomorphological and immunohistochemical evaluation of the influence of sex on parafollicular thyroid C cells in rats with chronic renal failure. The experiment included 40 Wistar rats after subtotal nephrectomy, after sham operation, and without any surgical procedure. Two weeks after nephrectomy, fragments of thyroids were collected from the examined animals. Paraffin sections were stained with H+E and by silver impregnation. Calcitonin (CT), synaptophysin (SPh), somatostatin (ST), and neuron-specific enolase (NSE) were detected immunohistochemically in C cells. In rats with experimental uremia, immunostaining for the examined substances increased significantly in comparison to the controls. We also observed higher number of C cells with a stronger reaction in the group of males, compared to the female rats.     

Regulation of estrogen receptor alpha by estradiol in pregnant and estradiol treated rats

Estrogens play an important role in tissue metabolism through specific regulation of several intracellular pathways. We studied ERalpha regulation in muscle and adipose tissue from pregnant and estradiol treated rats. In both groups, we identified three different ERalpha inmunoreactive proteins (80, 67 and 46 kDa) using total protein extracts. Because it has been showed that estrogens are able to promote rapid effects in several cellular models, we looked for three ERalpha-related proteins at plasma membrane. In skeletal muscle of both groups, we positively identified the three ERalpha-related isoforms in plasma membrane, but in adipose tissue from pregnant we were not able to identify ERalpha67, and in estradiol treated animals ERalpha80 was absent. Taking together, our results showed a tissue-specific regulation of whole-cell ERalpha-related proteins and ERalpha located at plasma membrane, which should be involved in non-genomic actions of 17beta-estradiol. The role of the three ERalpha inmunoreactive proteins is unknown, however, seems probably related to rapid activation of signalling pathways.     

Proliferation of lamellar whorls in arcuate neurons of the hypothalamus of male rats treated with estradiol benzoate or cyproterone acetate

Summary The arcuate nucleus of the hypothalamus (AH) of male rats which had been treated either with estradiol benzoate (E²B) or cyproterone acetate (CPA) was examined ultrastructurally for the presence of whorls of endoplasmic reticulum. The incidence of whorl containing neurons (WCN) was 2–4 times higher in the AH of animals treated for 2–3 weeks with E²B or for 2 weeks with CPA than in the AH of oil treated controls. CPA is a powerful anti-androgen while E²B acts both peripherally and centrally to limit testosterone production. These findings, together with previous evidence that whorls proliferate in AH of male rats deprived of androgen by morphine treatment or castration, suggest that steroid feedback (androgen alone or both androgen and estrogen) plays an important role in AH whorl proliferation. The possibility that WCN may be LH-RH containing neurons is suggested by the close correspondence between the number and location of WCN within AH as determined in this study and the distribution of LH-RH containing cells reported by others.The authors are indebted to Schering AG for supplying cyproterone acetate for this study. This work was supported by grants DA-00259, NS-09156 and MH-14677 from U.S.P.H.S.Research Scientist Development Award MH-38894Research Scientist Development Award MH-70180     

Insulin signaling in the liver and uterus of ovariectomized rats treated with estradiol

We used rat hepatic and uterine tissues to examine the impact of estradiol (E2) on insulin (INS) signaling. Ovariectomized (OVX) female Wistar rats were treated with E2 (20 microg/kg b.wt., i.p.) and used for the experiment 6h after E2 administration. To highlight E2 effects on tyrosine phosphorylation of INS receptor (IR) and INS receptor substrates (IRSs) and IRSs association with p85 subunit of phosphatidylinositol 3-kinase (PI3-K) in the context of INS signaling, E2-treated OVX rats were also injected with INS (20 IU, i.p.), 30 min before the experiment. Treatment with E2 did not change the levels of plasma INS and glucose (Glu). However, it significantly decreased the free fatty acid (FFA) level and increased uterine weight. Furthermore, the results show that E2 had no effect on the content of hepatic IR protein, but significantly increased IR protein content in the uterus and decreased IR tyrosine phosphorylation in both the liver and uterus. Compared to the control, hepatic IRS-1 and IRS-2 were significantly decreased and increased, respectively, after E2 treatment. Protein content of both molecules, IRS-1 and IRS-2, was increased in uterine tissue after E2 administration. Protein content of the p85 subunit of PI3-K and that of protein kinase B (Akt) were increased in the uterus, with no changes in the liver. The results suggest that E2 treatment induces tissue-specific changes in INS signaling. The consequences of E2 treatment on INS signaling molecules are more apparent in the uterus, but their physiological relevance for INS action is probably greater in the liver.     

The effect of orchidectomy on thyroid C cells and bone histomorphometry in middle-aged rats

This study was to evaluate the effect of androgen deficiency on thyroid immunoreactive C-cells and bone structure and function in a male orchidectomized middle-aged rat model. Fifteen-month-old male Wistar rats were divided into orchidectomized (Orx) and the sham-operated control (Sham) group. In the Orx group significant decreases (P < 0.05) were found in the volume of C cells (by 14%), their relative volume density (by 13%) and serum calcitonin concentration (by 54%) compared to the controls. Analyses of trabecular microarchitecture of the proximal tibia metaphysis showed that Orx induced marked decreases of cancellous bone area, trabecular thickness and trabecular number (by 52, 20 and 19% respectively; P < 0.05), whereas trabecular separation was increased by 27% (P < 0.05). In Orx rats, serum osteocalcin concentration was increased by 119% (P < 0.05), while serum calcium and phosphorus were 6 and 14% (P < 0.05) lower, respectively, compared to the levels in the Sham. In addition, urine calcium content was considerably higher (by 129%; P < 0.05) in Orx animals. These findings indicate that the androgen deficiency caused by Orx in middle-aged rats modulated the structure of C cells and diminished secretion of calcitonin. Histomorphometrical and biochemical analyses demonstrated a decrease of cancellous bone mass and increased bone turnover.     

Mounting behavior and lordosis behavior in castrated male rats treated with testosterone propionate, or with estradiol benzoate or dihydrotestosterone in combination with testosterone propinonate.

Treatment of prepuberally castrated male rats with testosterone propionate (TP, 50, 200, 500, or 1000 μg for 30 days) in adulthood stimulated the display of both mounting behavior and lordosis behavior. No correlation between mounting and lordosis behavior could be detected at any TP dose level. Treatment of prepuberally castrated male rats with either 1 μg estradiol benzoate (EB) or 500 μg dihydrotestosterone (DHT) for 60 days stimulated the display of mounting behavior in three of eight and four of eight rats, respectively. Treatment with 200 μg TP for the last 30 days of rats receiving either EB or DHT for 60 days resulted in an abrupt onset on mounting behavior as compared to rats treated with oil for 60 days. These results show additive effects of EB or DHT and TP upon mounting behavior by male rats and are interpreted as a support for the suggestion that testosterone to estrogen as well as testosterone to DHT conversion may be involved in the mechanism whereby testosterone activates the mounting behavior of castrated rats.     

Chronic treatment with estradiol restores depressive-like behaviors in female Wistar rats treated neonatally with clomipramine

Neonatal administration of clomipramine (CMI) induces diverse behavioral and neurochemical alterations in adult male rats that resemble major depression disorder. However, the possible behavioral alterations in adult female rats subjected to neonatal treatment with clomipramine are unknown. Therefore, the aim of this study was to explore the effect of neonatal treatment with CMI on adult female rats in relation to locomotion and behavioral despair during the estrus cycle. Also evaluated was the effect of chronic treatment with E2 on these female CMI rats. We found no effects on spontaneous locomotor activity due to neonatal treatment with CMI, or after 21 days of E2 administration. In the FST, neonatal treatment with CMI increased immobility and decreased active swimming and climbing behaviors. Influence of the ovarian cycle was detected only in relation to climbing behavior, as the rats in the MD phase displayed less climbing activity. Chronic E2 administration decreased immobility but increased only swimming in CMI rats. These results suggest that neonatal treatment with CMI induces despair-like behavior in female rats, but that chronic E2 administration generates antidepressant-like behavior by decreasing immobility and increasing swimming, perhaps through interaction with the serotonergic system.