The 21-aminosteroid antioxidant, U74389F, prevents estradiol-induced depletion of hypothalamic β-endorphin in adult female rats

A single intramuscular injection of 2 mg estradiol valerate (EV) results in neuronal degeneration and β-endorphin depletion in the hypothalamic arcuate nucleus of adult female rats. We have hypothesized that peroxidase-positive astrocytes in this brain region oxidize estrogens and catecholestrogens to semiquinone radicals which mediate oxidative neuronal injury. In the present study, dietary administration of the potent antioxidant 21-aminosteroid, U-74389F, completely blocked EV-induced β-endorphin depletion in the hypothalami of adult female rats. Neither EV nor 21-aminosteroid treatment had any effect on hypothalamic concentrations of neuropeptide Y and Met-enkephalin, confirming that the estradiol lesion is fairly selective for the β-endorphin cell population. The present findings support the hypothesis that the toxic effect of estradiol on hypothalamic β-endorphin neurons is mediated by free radicals.     

125I-抑制素在大鼠垂体和下丘脑的放射自显影研究

目的研究抑制素（INH）在体条件下能否通过大鼠的血脑屏障及在垂体或下丘脑的分布。方法采用颈静脉灌流和放射自显影技术，将20只SD大鼠分为4组，每组5只，第1～3组（实验组）颈静脉注射^125 I-INH50μl，第4（对照）组注射等量的生理盐水。第1，2，3组分别于注射后30，60和120min断头处死，取出垂体、下丘脑，以生理盐水洗涤，测量放射性计数，取放射性最大组的垂体与下丘脑组织行放射自显影分析。结果第1组垂体的放射性最高[（1008．00±5．78）Bq]，而第2和3组分别为（723．00±4．95）和（491．00±4．90）Bq；1～3组的下丘脑放射性分别为（20．00±1．01），（22．00±0．95）与（19．00±0．73）Bq。第4组垂体与下丘脑的放射性分别为（16．00±1．40），（15．00±0．98）Bq。各实验组大鼠垂体的放射性与对照组差异有统计学意义（P〈0．01），且在注射后30min放射性最大（第1组），60和120min后逐渐降低；而实验组与对照组大鼠的下丘脑放射性差异无统计学意义（P〉0．05）；放射自显影结果示，实验组大鼠的垂体组织上有明显的银颗粒，而对照组没有；实验组和对照组大鼠的下丘脑组织上均未见明显的银颗粒。结论^125I-INH能通过大鼠血脑屏障，垂体在注射后30min放射性最大，在大鼠垂体上有INH结合位点或受体，而在其下丘脑没有。     

下丘脑前部注射微量神经降压素对大鼠减压反应的影响

直接将神经降压素注射于大鼠下丘脑前部一侧减压区后,再以电刺激该区,观察其对减压反应的影响。结果表明,注射微量神经降压素后,可使电刺激该区所出现减压反应的最低值和持续时间,都比注射前明显增大。这证明,神经降压素参与电刺激大鼠下丘脑前部一侧减压区所出现的减压反应过程。     

Neonatal exposure to estradiol prevents the expression of ovarian hormone-induced luteinizing hormone and prolactin surges in adulthood but not antecedent changes in neuropeptide Y or adrenergic transmitter activity: Implications for sexual differentiation of gonadotropin secretion

Sex differences in adult patterns of mating behavior and gonadotropin secretion in rats are determined in part by the presence or absence of gonadal steroids during a perinatal critical period. For example, male rats and female rats exposed neonatally to androgen do not exhibit LH surge patterns when treated appropriately with ovarian hormones in adulthood, and there is evidence that this may be due to a failure of ovarian hormones to activate the hypothalamic neuronal systems that stimulate LH secretion in such animals. Because considerable evidence suggests that estradiol formed centrally from testosterone is responsible for the permanent defeminization of mating behavior and gonadotropin secretion, the present studies compared normal females with normal males and with females treated neonatally with estradiol on the ability of ovarian hormones to induce several important neurochemical changes antecedent to the LH surge, including changes in neuropeptide Y (NPY) and LH-releasing hormone (LHRH) concentrations in the median eminence, as well as changes in turnover rates for catecholamine transmitters in the medial basal hypothalamus and medial preoptic area. Normal ovariectomized female rats responded to sequential treatment with estradiol followed by progesterone with afternoon LH and prolactin (PRL) surges, and with sequential accumulation followed by decline in concentrations of LHRH and NPY in the median eminence prior to the LH surge. In addition, administration of progesterone increased the turnover rates of norepinephrine (NE) and epinephrine (EPI) in the arcuate-median eminence region of normal females. Gonadectomized male rats receiving the same ovarian hormone treatment failed to exhibit LH or PRL surges and displayed none of the changes in neurotransmitter turnover or peptide concentrations characteristically seen in the normal female. Unexpectedly however, when females that were treated with estradiol benzoate on days 1–3 postpartum were ovariectomized and treated with ovarian hormones in adulthood, they showed the same accumulation/decline in median eminence NPY concentrations and the same activation of NE and EPI turnover in the arcuate-median eminence region as normal females, even though they showed no LH or PRL surges or changes in median eminence LHRH concentrations. These results suggest that estradiol may not mediate all of the defeminizing actions of androgen exerted during the early neonatal period, and particularly those actions that result in a lack of responsiveness in central noradrenergic, adrenergic and NPY systems in adulthood. However, an action of neonatal estradiol may result in uncoupling of the LHRH neurosecretory system from normal excitatory neurochemical influences.     

The Changes in the Hypothalamo-Pituitary-Gonadal Axis of Streptozotocin-Treated Male Rats Depend from Age at Diabetes Onset

The influence of age at diabetes onset and of capillary microangiopathy on the severity and evolution of hypothalamo-pituitary-gonadal changes was studied morphologically and morphometrically in male rats 4 and 8 months after streptozotocin injection. At each time period we studied 2 groups of rats, one made diabetic before (age 1 month), the other after puberty (age 3 months), and compared them with corresponding controls. The size of hypothalamic axons, numerical density and size of pituitary gonadotrophs, size of testicular tubules, and basement membrane thickness of retinal capillaries were measured. Major differences were found at 8 months. Changes of pituitary glands (i.e. small and numerous gonadotrophs) and testes (i.e. small tubular size) were more important in pre- than in postpubertal diabetic rats. This was a consequence of the aggravating prepubertal diabetes between 4 and 8 months. On the contrary, these changes partially regressed in postpubertal diabetic animals. Pituitary and testicular changes were correlated. Other lesions, such as swollen axonal processes in the hypothalamus, increased thickness of seminiferous epithelium and of capillary basement membranes, though very evident in diabetics, were independent from age at induction. Neither microangiopathy nor glycemia were correlated with any other change which confirmed their secondary role in diabetic neuroendocrine disorders. Thus, two types of diabetic disorders of the hypothalamo-pituitary-gonadal axis could be distinguished: 1) those with irreversible effects on immature yet partially reversible effects on mature structures; and 2) those independent from age at induction.     

Increased feeding and neuropeptide Y (NPY) but not NPY mRNA levels in the hypothalamus of the rat following central administration of the serotonin synthesis inhibitorp-chlorophenylalanine

Neurons containing serotonin (5-HT), a potent anorexic agent, come into contact with neuropeptide Y-ergic neurons, that project from the arcuate nucleus (ARC) to the paraventricular nucleus (PVN). NPY powerfully stimulates feeding and induces obesity when injected repeatedly into the PVN. We hypothesize that 5-HT tonically inhibits the ARC-PVN neurons and that balance between the two systems determines feeding and energy homeostasis. This study aimed to determine whether central injection of the 5-HT synthesis inhibitor p-chlorophenylalanine (pCPA), which increases feeding, increased hypothalamic NPY and NPY mRNA levels. pCPA (10 mg/kg in 3 μl) was administered into the third ventricle either as a single injection (n = 8) or daily for 7 days (n = 8). Control rats received a similar injection of saline. pCPA significantly increased food intake compared with controls after both single and repeated injections (P < 0.05). NPY levels were measured by radioimmunoassay in microdissected hypothalamic extracts. NPY levels in the acutely treated group were significantly increased in the paraventricular nucleus (PVN; by 41%,P = 0.01), anterior hypothalamic area (AHA; by 34%,P < 0.01) and lateral hypothalamic area (LHA; by 41%,P < 0.02). In the 7-day-treated group, NPY levels were also increased in the same areas, i.e. PVN (by 24%,P < 0.01), AHA (by 30%,P < 0.01) and LHA (by 38%,P = 0.01). There were no significant changes in the ARC or any other region or in hypothalamic NPY mRNA levels. pCPA administration increased NPY levels in several regions notably the PVN. This is a major site of NPY release, where NPY injection induces feeding. We suggest that the hyperphagia induced by pCPA is mediated by increased NPY levels and secretion in the PVN. This is further evidence for interactions between NPY and 5-HT in the control of energy homeostasis.     

Hypothalamic knife-cut hyperphagia and obesity in weanling rats

In previous reports weanling female rats fed a high-fat diet had a delayed response to hypothalamic knife cuts. In the present report similar cuts in similar rats fed a standard low fat diet became overweight without delay, suggesting that dietary fat is a critical variable in juvenile onset obesity. Adult rats given knife-cuts comparable to those in the weanlings gained weight far more rapidly and achieved higher weights than did those cut as weanlings, suggesting that appetite modulating axons can develop after weaning. Finally, group vs single housing did not influence the age of onset or the magnitude of knife-cut obesity.     

Afferent fiber connections from lower brain stem to hypothalamus studied by the horseradish peroxidase method with special reference to noradrenaline innervation

Summary Attempts were made to determine the afferent projections to the anterior hypothalamus including the preoptic area from the lower brain stem by means of the horseradish peroxidase method combined with monoamine oxidase staining to identify noradrenaline (NA) neurons. In addition to this technique, a histofluorescence analysis was performed. NA fibers in the medial part of the anterior hypothalamus were mainly supplied by A1 and A2 NA neuron groups, while the lateral part and periventricular zone received NA terminals from both pontine and medulla oblongata NA neuron groups. Furthermore, the present study indicated that there were direct projections to the anterior hypothalamus from non-noradrenergic neurons in the lower brain stem: nuclei raphe dorsalis, centralis superior, cells in the mesencephalic and pontine central gray matter, nuclei parabrachialis lateralis and medialis, cells around fasciculus longitudinalis medialis.Abbreviations CA Commissura anterior - CO Chiasma opticum - DP Decussatio pyramidum - DPCS Decussatio pedunculorum cerebellarium superiorum - F Columna fornicis - FLM Fasciculus longitudinalis medialis - FMT Fasciculus mamillothalamicus - GCM Griseum centrale mesencephali - GCP Griseum centrale pontis - LL Lemniscus lateralis - LM Lemniscus medialis - PCM Pedunculus cerebellaris medius - PCS Pedunculus cerebellaris superior - TO Tractus opticus - TS Tractus solitarius - TVme Tractus mesencephalicus nervi trigemini - V Ventriculus tertius - VTS Tractus spinalis nervi trigemini - am nucleus ambiguus - B Barrington nucleus - com nucleus commissuralis - cp nucleus caudatus putamen - cs nucleus centralis superior - ct nucleus corporis trapezoidei - cu nucleus cuneatus - dX nucleus dorsalis nervi vagi - Gd nucleus tegmentalis dorsalis (von Gudden) - gr nucleus gracilis - Gv nucleus tegmentalis ventralis (von Gudden) - ha nucleus hypothalamicus anterior - hl nucleus hypothalamicus lateralis - hpe nucleus periventricularis (hypothalami) - hvm nucleus ventromedialis hypothalami - lc nucleus locus coeruleus - oi nucleus olivaris inferior - p nucleus pontis - pa nucleus paraventricularis - pbl nucleus parabrachialis lateralis - pbm nucleus parabrachialis medialis - ph nucleus praepositus hypoglossi - pol nucleus preopticus lateralis - pom nucleus preopticus medialis - pop nucleus preopticus periventricularis - rd nucleus raphe dorsalis - re nucleus reuniens - rl nucleus reticularis lateralis - rm nucleus raphe magnus - ro nucleus raphe obscrus - sc nucleus suprachiasmaticus - so nucleus supraopticus - st nucleus interstitialis striae terminalis - td nucleus tractus diagonalis (Broca) - ts nucleus tractus solitarii - Vme nucleus mesencephalicus nervi trigemini - Vmo nucleus motorius nervi trigemini - Vts nucleus tractus spinalis nervi trigemini - XII nucleus nervi hypoglossi     

Hypothalamic sensitivity to 2-deoxy-D-glucose and glucose: effects on feeding behavior

The effect on food intake of intrahypothalamic deposition of glucose or the glucose metabolic inhibitor 2-deoxy-D-glucose was studied to determine the function of the often hypothesized hypothalamic glucoreceptors in feeding behavior. Our results suggest that the lateral hypothalamic region, as well as the medial hypothalamus, contain glucoreceptive cells that form part of a feeding system.