RESUMO
The authors examined the autopsied brains from three patients who had a fatal hyperthermia syndrome. There was marked hypothalamic noradrenaline depletion in all three patients, severe brain choline acetyltransferase deficiency with nucleus basalis cell loss in two patients, and mild to moderate brain choline acetyltransferase loss in one patient. Striatal dopamine metabolite/dopamine ratio was below normal in two patients and not elevated, as would be expected after short-term neuroleptic administration, in the third. This suggests that reduced capability (aggravated by the cholinergic deficit) of the nigrostriatal dopamine system to respond adequately to stress and/or neuroleptic-induced receptor blockade may be important in the development and course of fatal hyperthermia syndrome.
Assuntos
Química Encefálica , Catatonia/metabolismo , Colina O-Acetiltransferase/análise , Dopamina/análise , Febre/metabolismo , Síndrome Maligna Neuroléptica/metabolismo , Norepinefrina/análise , Adolescente , Adulto , Autopsia , Encéfalo/enzimologia , Encéfalo/metabolismo , Encéfalo/patologia , Catatonia/patologia , Corpo Estriado/análise , Corpo Estriado/patologia , Feminino , Febre/patologia , Humanos , Hipotálamo/análise , Hipotálamo/patologia , Masculino , Pessoa de Meia-Idade , Síndrome Maligna Neuroléptica/patologia , Substância Inominada/análise , Substância Inominada/patologia , SíndromeRESUMO
A novel neuropeptide with 38 residues (PACAP38) was isolated from ovine hypothalamic tissues using the pituitary adenylate cyclase activation in rat pituitary cell cultures as a parameter of the biological activity (Miyata et al, Biochem. Biophys. Res. Commun. 164, 567-574, 1989). From the side fractions obtained during the purification of PACAP38, a shorter form peptide with 27 residues corresponding to the N-terminal 27 amino acids of PACAP38 and amidated C-terminus was isolated and named as PACAP27. Synthetic PACAP27 showed a biological activity of adenylate cyclase stimulation comparable to PACAP38. Moreover PACAP27 which shows a considerable homology with vasoactive intestinal polypeptide (VIP) demonstrated a similar vasodepressor activity as VIP, but the adenylate cyclase stimulating activity was about 1000 times greater than VIP.
Assuntos
Adenilil Ciclases/isolamento & purificação , Neuropeptídeos/isolamento & purificação , Neuropeptídeos/metabolismo , Peptídeos/metabolismo , Hipófise/enzimologia , Sequência de Aminoácidos , Animais , Células Cultivadas , Ativação Enzimática , Hipotálamo/análise , Dados de Sequência Molecular , Peptídeos/síntese química , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase , Ovinos , Peptídeo Intestinal Vasoativo/metabolismoRESUMO
We recently reported isolation, characterization and synthesis of a novel ovine hypothalamic peptide with 38 residues which stimulates accumulation of cAMP in rat anterior pituitary cell cultures. The peptide was named PACAP38 (pituitary adenylate cyclase-activating polypeptide with 38 residues). The presence of another peptide corresponding to the N-terminal 1-27 residues (PACAP27) was also demonstrated. Both PACAP38 and PACAP27 have an amidated C-terminus. Antisera against synthetic PACAP27 were generated in rabbits. These antisera were tested for titer and specificity in enzyme-linked immunosorbent assay. One of the antisera (no. 88121-3) exhibited a high titer of antibody, which was specific to PACAP27 and PACAP38 with exception of slight cross-reactivity with ovine CRF (oCRF). Therefore, the antibodies against oCRF were removed from the antiserum using a solid phase method. Removal of oCRF antibodies was confirmed by enzyme-linked immunosorbent assay. A dense immunoreactive fiber network was found in both external and internal zones of the median eminence and pituitary stalk. The fibers were demonstrated to be in close contact with the hypophysial portal capillaries. The preabsorption of antiserum with vasoactive intestinal polypeptide or with the mixture containing TRH, LHRH, oCRF, ovine GH-releasing factor, somatostatin, and bovine thyroglobulin did not affect the immunostaining. On the other hand, the preabsorption of antiserum with an excess of PACAP27 or PACAP38 abolished the immunostaining. Therefore, the staining is considered specific for PACAP27 and PACAP38. Stained fibers were also present in the posterior pituitary. A dense fiber network was observed and the lateral hypothalamus the fibers appeared to cling to unstained neuronal cell bodies and their dendrites. In the lateral septum the fibers surrounded some blood vessels. Immunolabeled cell bodies were found in the paraventricular and supraoptic nuclei. These findings support the view that PACAP may play a multifunctional role, including that of a hypophysiotropic hormone, neurotransmitter, neuromodulator, and vasoregulator.
Assuntos
Hipotálamo/análise , Neuropeptídeos/análise , Sequência de Aminoácidos , Animais , Especificidade de Anticorpos , Ensaio de Imunoadsorção Enzimática , Histocitoquímica , Soros Imunes/análise , Soros Imunes/imunologia , Técnicas Imunoenzimáticas , Masculino , Dados de Sequência Molecular , Neuropeptídeos/imunologia , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase , Hipófise/análise , OvinosRESUMO
Hyperphagia and obesity are often associated, and the origins of the biochemical modifications leading to these syndromes might be in the hypothalamus. Indeed, food intake is regulated by numerous neuropeptides in various hypothalamic nuclei, including the paraventricular (PVN), arcuate (ARC), ventromedian (VMN) and suprachiasmatic (SCH) nuclei. Among these peptides, neuropeptide Y (NPY) is the most potent inducer of food intake whereas neurotensin (NT) decreases food intake. We measured these two peptides in microdissected hypothalamic nuclei in obese Zucker rats that ate 30% more food than their lean counterparts. Neuropeptide Y and neurotensin levels varied in opposite directions: In the hyperphagic obese Zucker rats, the NPY concentrations were significantly greater than those in the lean normophagic rats in the ARC (+30%), PVN (+60%) and SCH (+94%) nuclei, whereas the NT levels were significantly lower in the ARC (-40%), PVN (-31%) VMN (-66%) and SCH (-47%) nuclei. Both these variations tend to increase food intake. Feeding periodicity might also be modified because large variations of the two peptides have been measured in the supra-chiasmatic nucleus, which is considered the most important regulator of feeding rhythm. The results reinforce the hypothesis that hyperphagia in obesity is associated with a biochemical modification in the central nervous system because the peripheral status of NT and NPY was not modified in the obese rats. Because levels of other hypothalamic peptides, such as opioid peptides and somatostatin, are also slightly modified, it can be concluded that hyperphagia in obesity is associated with a central peptidergic dysregulation. Research on drugs reacting specifically with the receptor of these peptides might have interesting implications for the treatment of hyperphagia and, therefore, of obesity.
Assuntos
Hiperfagia/etiologia , Hipotálamo/fisiopatologia , Neuropeptídeo Y/fisiologia , Neurotensina/fisiologia , Obesidade/complicações , Animais , Núcleo Arqueado do Hipotálamo/análise , Núcleo Arqueado do Hipotálamo/fisiopatologia , Modelos Animais de Doenças , Hiperfagia/complicações , Hipotálamo/análise , Neuropeptídeo Y/análise , Neurotensina/análise , Núcleo Hipotalâmico Paraventricular/análise , Núcleo Hipotalâmico Paraventricular/fisiopatologia , Ratos , Ratos Zucker , Núcleo Hipotalâmico Ventromedial/análise , Núcleo Hipotalâmico Ventromedial/fisiopatologiaRESUMO
Beef cows (n = 64) were slaughtered to evaluate effects of dietary energy and calf removal (CR) on hypothalamic and adenohypophysial endocrine characteristics. From d 190 of gestation until parturition, cows received maintenance (ME; n = 32) or low (LE; n = 32) energy diets (ME = 100%, LE = 70% NRC recommendations). After parturition, half (n = 16) of each prepartum diet group received low (LE; n = 32) or high (HE = 130% NRC; n = 32) energy diets. At 30 d postpartum, cows were slaughtered 0 or 48 hr after CR. Hypothalami [preoptic area (POA), hypothalamus (HYP), stalk-median eminence (SME)] and pituitaries were collected. Basal and K(+)-induced release of GnRH from SME, and pituitary luteinizing hormone (LH) and follicle stimulating hormone (FSH) did not differ among groups (P greater than .05). Hypophyseal LH was correlated (P less than .01) with body condition score (BCS) at parturition and slaughter (r = .36 and .47, respectively). Prepartum LE diet increased (P less than .05) met-enkephalin in POA compared to prepartum ME (.59 +/- .05 vs. .44 +/- .04 pmol/mg) regardless of postpartum diet or suckling status. Concentrations of beta-endorphin in combined HYP + POA were decreased (P less than .05) by 48 hr CR (15.1 +/- 1.1 vs. 18.1 +/- 0.7 fmol/mg).(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Bovinos/metabolismo , Dieta , Endorfinas/análise , Gonadotropinas Hipofisárias/análise , Hormônios Liberadores de Hormônios Hipofisários/análise , Animais , Ingestão de Energia , Encefalina Metionina/análise , Feminino , Hormônio Foliculoestimulante/análise , Hipotálamo/análise , Lactação , Hormônio Luteinizante/análise , Hipófise/análise , beta-Endorfina/análiseRESUMO
Twenty-one castrated oestrogen-primed Wistar rats, which were 2-months-old, were injected via the jugular vein with 100 mu Ci/100 g body weight of [3H]RU 486 or [3H]progesterone. Some of these received unlabelled compounds for competition studies. Samples of reproductive tract, pituitary and hypothalamus were excised after 15 min. The 4-microns frozen sections were processed for thaw-mounted autoradiography. The exposure time of the autoradiogram was approximately 6 months. After the injection of [3H]RU 486 and [3H]progesterone, the nuclear concentration of radioactivity was most distinct in muscular and stromal cells of the uterus, and the epithelial nuclei of lumina and glands showed weak labelling. Nuclear localization was also observed in muscle cells of the vagina, cervix and oviduct. After injection of [3H]progesterone, the radioactivity was found in the nuclei and cytoplasm of anterior pituitary cells and some cells showed a preferential nuclear concentration of radioactivity. The distribution of [3H]RU 486 in the anterior pituitary was more extensive than that of [3H]progesterone. In the hypothalamus, specific localization of [3H]RU 486 and [3H]progesterone existed in neurones accumulated in the preoptic nucleus, preoptic suprachiasmatic nucleus and the periventricular nucleus. No localization was found in the diaphragm. Pretreatment with RU 486, but not with dexamethasone, reduced the nuclear concentration of radioactivity of [3H]progesterone in the vagina, uterus, oviduct, pituitary and hypothalamus. The nuclear concentration of radioactivity after injection of [3H]RU 486 was also decreased by preinjection with progesterone. The autoradiographic results suggest that RU 486 and progesterone competed for the specific binding site (possibly a progesterone receptor) in the target cells at the levels of the uterus, pituitary and hypothalamus in vivo.
Assuntos
Hipotálamo/análise , Mifepristona/análise , Hipófise/análise , Progesterona/análise , Útero/análise , Animais , Autorradiografia , Núcleo Celular/análise , Citoplasma/análise , Dexametasona/farmacologia , Epitélio/análise , Epitélio/ultraestrutura , Feminino , Hipotálamo/ultraestrutura , Mifepristona/farmacocinética , Músculos/análise , Músculos/ultraestrutura , Ovariectomia , Hipófise/ultraestrutura , Adeno-Hipófise/análise , Progesterona/farmacocinética , Progesterona/farmacologia , Ratos , Ratos Endogâmicos , Distribuição Tecidual , Trítio , Útero/ultraestruturaRESUMO
Catecholamine content was studied in hypothalamus of neonatal Wistar female rats treated with 4-hydroxyestradiol-17 (4-OH-E2) in a dose of 10 mg for 1-5 days of life. 4-OH-E2 induced a reliable increase in hypothalamic noradrenaline level in 24 h after the last injection, but not on the 7th, 10th or 12th postnatal days. There was no change in dopamine level. We have postulated a relationship between the increase in hypothalamic noradrenaline content induced by 4-OH-E2 and defeminization effects of 4-OH-E2 on the developing brain of female rats.
Assuntos
Estradiol/análogos & derivados , Estrogênios de Catecol/farmacologia , Hipotálamo/análise , Norepinefrina/análise , Fatores Etários , Animais , Animais Recém-Nascidos , Estradiol/farmacologia , Feminino , Ratos , Ratos EndogâmicosRESUMO
By means of the indirect immunohistochemical method distribution of S-100 containing cells has been studied in sections of the mediobasal hypothalamus (astrocytes) and adenohypophysis (follicular-stellate cells) in newborn, 10- and 21-day-old rats under normal development and under protein insufficiency. For this the animals are given the diet containing 6% of protein (control--25% of protein). S-100 containing cells are revealed in the hypothalamus and adenohypophysis in 10- and 21-day-old animals. In the brain of the newborn rats S-100 immunoreactive cells are not revealed. At the ultrastructural level the diaminobenzidine (DAB) reaction products in the immunoreactive cells are revealed diffusely along the whole cytoplasm of the cells, in nuclei the DAB reaction products are absent. Part of S-100 containing cells is essentially lowered, comparing with the control. In the rat adenohypophysis part of S-100 containing cells from the 10th up to the 21st day also decreases. Unlike the hypothalamus, however, content of cells, immunopositive to S-100 exceeds the analogous index in the control rats of the corresponding age groups.
Assuntos
Hipotálamo/análise , Adeno-Hipófise/análise , Deficiência de Proteína/metabolismo , Proteínas S100/análise , Fatores Etários , Animais , Hipotálamo/ultraestrutura , Imuno-Histoquímica , Microscopia Eletrônica , Adeno-Hipófise/ultraestrutura , Ratos , Ratos Endogâmicos , Proteínas S100/ultraestruturaRESUMO
Galanin is a widely distributed regulatory peptide which modulates the pituitary secretion of PRL and GH. Estrogen administration strongly stimulates galanin gene expression in the rat anterior pituitary. In adult female Fischer 344 rats, estrogen also induces hyperplasia of lactotropes. We used immunocytochemical analysis to assess the effects of estrogen on galanin-like immunoreactivity (Gal-IR) in the rat pituitary and hypothalamus during sc diethylstilbestrol (DES) implantation and after its removal at 30 days. In the anterior pituitary, DES implantation increased the portion of Gal-IR-containing cells from less than 2% in the control rats to 18.3% after 3 days of DES and 36% after 30 days. These changes paralleled the lactotrope hyperplasia exhibited in response to DES exposure. Ten and 30 days after removal of the DES capsules, the percentage of Gal-IR-containing cells in the anterior pituitary decreased to 6.3% and 1.5%, respectively. Colocalization studies revealed that Gal-IR-containing cells were predominantly lactotropes. Immunoelectron microscopy demonstrated that Gal-IR was concentrated in the Golgi region of these hyperplastic lactotropes and suggests that little of the synthesized galanin is secreted. The distribution of Gal-IR in the hypothalamus, median eminence, and neurohypophysis was unaffected by DES treatment. These data demonstrate that galanin is synthesized by hyperplastic pituitary lactotropes of Fischer 344 rats and that peptide accumulation is dependent on the presence of circulating estrogens. In contrast, neuronal galanin synthesis in the hypothalamus does not appear to be regulated by estrogen.
Assuntos
Dietilestilbestrol/farmacologia , Peptídeos/metabolismo , Adeno-Hipófise/metabolismo , Prolactina/metabolismo , Animais , Grânulos Citoplasmáticos/análise , Grânulos Citoplasmáticos/ultraestrutura , Feminino , Galanina , Complexo de Golgi/análise , Hormônio do Crescimento/análise , Hormônio do Crescimento/metabolismo , Hiperplasia , Hipotálamo/análise , Técnicas Imunoenzimáticas , Eminência Mediana/análise , Microscopia Eletrônica , Peptídeos/análise , Adeno-Hipófise/patologia , Adeno-Hipófise/ultraestrutura , Neuro-Hipófise/análise , Prolactina/análise , Ratos , Ratos Endogâmicos F344 , Distribuição TecidualRESUMO
An interaction between the immune and endocrine systems has been long known. This association is further strengthened by the finding that splenic lymphocytes have the capacity to produce molecules similar to if not the same as classical hormones, including several members of the opiate family, PRL, GH, and neuropeptide Y. Because of such findings and because of information from other laboratories suggesting that LHRH might have direct effects upon the immune system, we hypothesized that immune cells themselves might contain LHRH. Lymphocytes were purified from spleens of intact adult male Sprague-Dawley rats and the cells were lysed with sodium hydroxide. The concentration of immunoreactive LHRH was 403 +/- 184 pg/20 X 10(6) lymphocytes. Increasing amounts of lymphocyte lysate displaced [125-I]LHRH from LHRH antibody in a manner parallel to that produced by synthetic hypothalamic LHRH, suggesting immunologic similarity between lymphocyte and hypothalamic LHRH. Lymphocyte LHRH-like immunoactivity coeluted from Nova-Pak C18 columns with synthetic hypothalamic LHRH. When lymphocyte lysates were applied to rat anterior pituitary cells in monolayer culture, significant stimulation of LHRH secretion was seen, from 2,144 +/- 54 pg LH/ml.4 h to 15,364 +/- 587 pg LH/ml.4 h (P less than 0.001), a finding verified in five additional experiments. In other studies, this LH response evoked by lymphocyte lysates was found to be dose dependent and could be significantly inhibited by an LHRH-antagonist. Furthermore, when lymphocyte lysate and identically treated synthetic LHRH were HPLC fractionated, there was coelution of lysate and hypothalamic LHRH bioactivity. The lysate itself contained no substantial LH immunoreactivity. Thus, lymphocytes from spleens of adult male rats contain an immunoactive and bioactive LHRH, a finding further strengthening an association between the endocrine and immune systems.
Assuntos
Hormônio Liberador de Gonadotropina/análise , Linfócitos/análise , Baço/citologia , Animais , Bioensaio , Cromatografia Líquida de Alta Pressão , Hipotálamo/análise , Hormônio Luteinizante/metabolismo , Masculino , Adeno-Hipófise/efeitos dos fármacos , Adeno-Hipófise/metabolismo , Radioimunoensaio , Ratos , Ratos EndogâmicosRESUMO
Two molecular forms of gonadotropin-releasing hormone (GnRH) were demonstrated in hypothalamic extracts of M. domestica using high performance liquid chromatography and radioimmunoassay with specific GnRH antisera. One form eluted in the same position as synthetic mammalian GnRH and was quantified equally by two mammalian GnRH antisera, while the second form coeluted with synthetic chicken GnRH II and was quantified equally with two chicken GnRH II antisera. The finding of chicken GnRH II in a South American species of marsupial, which has previously been reported in some Australian species of marsupial and in species of Aves, Reptilia, Amphibia, Osteichthyes and Chondrichthyes, supports our hypothesis that this widespread structural variant may represent an early evolved and conserved form of GnRH.
Assuntos
Hormônio Liberador de Gonadotropina/análogos & derivados , Marsupiais/metabolismo , Hormônios Liberadores de Hormônios Hipofisários/análise , Sequência de Aminoácidos , Animais , Cromatografia Líquida de Alta Pressão , Hormônio Liberador de Gonadotropina/análise , Hipotálamo/análise , Mamíferos , Dados de Sequência Molecular , Radioimunoensaio , América do SulRESUMO
Mouse monoclonal antibody (mAb) L6 identifies an antigen expressed on the cell surface of many different human carcinomas. While studying the binding activity of mAb L6 to intracerebral tumor xenografts of human lung carcinoma LX-1 cells in nude rats using immunohistological techniques, we observed that L6 can also bind to a cytoplasmic antigen expressed in the magnocellular component of the hypothalamo-neurohypophysial system. Double-labeling experiments with antisera to vasopressin and oxytocin confirmed the localization of L6 immunoreactivity within both peptide-containing cell groups. L6 immunoreactivity in Brattleboro rats (with genetic deletion in the vasopressin gene) was exclusively localized within oxytocin neurons. Oxytocin and vasopressin failed to block L6 staining which suggested that its target epitope resides within the neurophysin sequence, and this explanation was supported by the finding that adsorption of L6 with porcine neurophysin completely eliminated hypothalamic immunoreactivity. Western blot analysis of bovine neurophysin and human pituitary extracts identified L6-immunoreactive bands which corresponded to the position of neurophysin and pro-pressophysin, confirming that L6 immunoreactivity in hypothalamus is related to neurophysin. Thus, monoclonal antibody L6, which is highly reactive with a membrane antigen of human lung cancer cell line LX-1, recognizes a cytoplasmic epitope in hypothalamic neurons identified as neurophysin by immunohistochemistry and Western analysis.
Assuntos
Antígenos de Neoplasias/imunologia , Antígenos de Superfície/imunologia , Hipotálamo/análise , Neurofisinas/análise , Adsorção , Animais , Anticorpos Monoclonais , Western Blotting , Eletroforese em Gel de Poliacrilamida , Humanos , Técnicas Imunoenzimáticas , Ocitocina , Ratos , Ratos Nus , Células Tumorais Cultivadas , VasopressinasRESUMO
Effects of the glucocorticoid milieu on the basal and ether stress-induced prolactin (PRL) release and on the immunostaining for hypothalamic vasoactive intestinal peptide (VIP), beta-endorphin (beta-EP), dynorphin-A (DYN-A) and methionine-enkephalin (Met-ENK), were examined in separate groups of male rats. After colchicine treatment in intact rats, VIP-containing cell bodies were observed only in the suprachiasmatic nucleus (SCN). Adrenalectomy (ADX), performed 7 days previously, resulted in the additional appearance of VIP-immunoreactive neurons in the parvocellular subdivision of the paraventricular nucleus (PVN), as well as in significantly higher basal and stressed PRL levels than intact values. Treatment of intact rats with a high dose (500 micrograms/kg body weight (s.c.) daily for 7 days) of dexamethasone (DEX), but not with a low dose (50 micrograms/kg) of DEX, significantly reduced both the basal and stressed PRL release. Administration of either the low or high dose of DEX to ADX rats prevented the appearance of the PVN-VIP neurons. In addition, the ADX-induced high basal and stressed PRL levels were restored to intact values by the low dose of DEX, and completely suppressed by the high dose of DEX. The staining of SCN-VIP-, beta-EP-, DYN-A or Met-ENK neurons was not affected by any treatment employed in this study. These results suggest that the appearance of PVN-VIP immunostaining in ADX rats may, at least in part, be responsible for the enhanced PRL secretion observed in this group. However, SCN-VIP-, beta-EP-, DYN-A- or Met-ENK neurons do not seem to play a pivotal role in the glucocorticoid regulation of PRL secretion.
Assuntos
Dinorfinas/análise , Encefalina Metionina/análise , Glucocorticoides/farmacologia , Hipotálamo/análise , Prolactina/metabolismo , Peptídeo Intestinal Vasoativo/análise , beta-Endorfina/análise , Adrenalectomia , Animais , Dexametasona/administração & dosagem , Dexametasona/farmacologia , Relação Dose-Resposta a Droga , Éteres , Técnicas Imunológicas , Masculino , Ratos , Núcleo Supraquiasmático/efeitos dos fármacos , Núcleo Supraquiasmático/metabolismo , Núcleo Supraquiasmático/ultraestruturaRESUMO
Maharishi-4 (M-4), an ayurvedic food supplement, was tested for anticarcinogenic and anticancer properties against 7,12-dimethylbenz(a)anthracene (DMBA)-induced mammary tumors in rats. The 6% M-4-supplemented diet protected DMBA-induced carcinogenesis by reducing both tumor incidence and multiplicity during initiation and promotion phases. The control animals who developed tumors when supplemented with M-4 diet for four weeks showed tumor regression in 60% of cases. There was no significant difference in the food intake or weight gain in rats who were on M-4-supplemented diet compared to control group. Possible mechanisms of action of M-4 are discussed.
Assuntos
Adenocarcinoma/prevenção & controle , Antineoplásicos Fitogênicos , Neoplasias Mamárias Experimentais/prevenção & controle , Ayurveda , Plantas Medicinais , 9,10-Dimetil-1,2-benzantraceno , Adenocarcinoma/induzido quimicamente , Adenocarcinoma/tratamento farmacológico , Animais , Encefalina Metionina/análise , Estradiol/sangue , Feminino , Hipotálamo/análise , Neoplasias Mamárias Experimentais/induzido quimicamente , Neoplasias Mamárias Experimentais/tratamento farmacológico , Hipófise/análise , Prolactina/sangue , Ratos , Ratos Endogâmicos , beta-Endorfina/análiseRESUMO
We have completely purified, in parallel, a low molecular weight, non-specific, non-lipidic, Na+,K(+)-ATPase inhibitory factor from bovine hypothalamic and pituitary tissues. In the final purification step we obtain, from both tissues, a single, homogeneous peak, with a maximal absorbance at 247 nm. This factor, at physiological concentrations of potassium (5-25 mM), inhibits in a dose-response manner Na+,K(+)-ATPase and displaces ouabain from its receptor at the enzyme structure. The factor isolated from both tissues is identical, being the specific activity per weight of tissue higher in hypophysis. No factor was found in cerebral cortex, used as tissue control.
Assuntos
Córtex Cerebral/análise , Hipotálamo/análise , Hipófise/análise , ATPase Trocadora de Sódio-Potássio/antagonistas & inibidores , Animais , Ligação Competitiva , Bovinos , Concentração de Íons de Hidrogênio , Hidrólise , Peso Molecular , Ouabaína/metabolismo , Soroalbumina Bovina/farmacologia , Fosfolipases Tipo C/metabolismoRESUMO
The changes in adrenocorticotropin (ACTH) release before, during and after sympathetic nerve degeneration following superior cervical ganglionectomy (SCGx) were examined in male rats. A 12-fold increase of circulating ACTH was found in both SCGx and sham-operated rats 6 h after surgery. In sham-operated rats, plasma ACTH decreased by about half 16-22 h after surgery, whereas in SCGx rats it remained at a high concentration from 16 to 54 h after surgery, attaining basal values by 120 h post-SCGx. In SCGx rats, MBH corticotropin-releasing hormone (CRH) content decreased significantly from 16 to 54 h after surgery, while in controls it remained unmodified. Significantly smaller arginine vasopressin (AVP) contents were found in MBH of SCGx rats as compared to sham-operated controls, 16-54 h after surgery. In rats exposed to ether or immobilization stress 22 h after SCGx, plasma ACTH levels were significantly higher than in controls; however, since unstressed ACTH levels were about twice as high in SCGx rats, the percent increase of ACTH was smaller in the SCGx group. A decreased response of plasma ACTH to ether or immobilization stress was found in rats 7 days after SCGx. In rats subjected to a simultaneous adrenalectomy (Adx) and SCGx or sham-SCGx, plasma ACTH levels increased to a similar extent in both groups. ACTH increase after Adx was accompanied by decreases in MBH CRH, and absence of significant changes in MBH AVP contents. Rats subjected to pinealectomy (Px) or sham-Px 1 week earlier and killed 22 h earlier exhibited similar responses in plasma ACTH and MBH CRH to SCGx regardless of pineal intactness.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Hormônio Adrenocorticotrópico/metabolismo , Ganglionectomia , Degeneração Neural/fisiologia , Neurônios/patologia , Degeneração Walleriana/fisiologia , Hormônio Adrenocorticotrópico/sangue , Animais , Arginina Vasopressina/análise , Hormônio Liberador da Corticotropina/análise , Gânglios Simpáticos/citologia , Gânglios Simpáticos/fisiologia , Hipotálamo/análise , Masculino , Ratos , Ratos EndogâmicosRESUMO
In order to evaluate the effects of environmental lighting on the hypothalamic secretion of luteinizing hormone-releasing hormone (LHRH), intact and castrated male Wistar rats were kept either in periodic (LD) or in constant light (LL) for 1 week, and the hypothalamic LHRH concentration was measured by radioimmunoassay. In the intact rats the LHRH concentration was lower in constant than in periodic light (30 +/- 3 vs 61 +/- 3 pg mg-1, P less than 0.05), whereas in the castrated rats it was higher (16 +/- 2 vs 5 +/- 1 pg mg-1, P less than 0.05). The intact and castrated rats received melatonin (50 micrograms) or saline injections daily at either 09.00 h or 16.00 h in LD and LL for 1 week. In intact or castrated rats the melatonin injections were not able to prevent the change of hypothalamic LHRH in constant light. In castrated rats testosterone (125 or 250 micrograms) had no effect on the LHRH concentration under either lighting condition. The present results suggest that constant light reduces the synthesis of hypothalamic LHRH in intact rats but increases it in castrated rats. These effects of constant light do not appear to be related to melatonin.
Assuntos
Hormônio Liberador de Gonadotropina/análise , Hipotálamo/análise , Luz , Melatonina/farmacologia , Testosterona/farmacologia , Animais , Relação Dose-Resposta a Droga , Hipotálamo/efeitos dos fármacos , Masculino , Melatonina/administração & dosagem , Orquiectomia , Periodicidade , Ratos , Ratos EndogâmicosRESUMO
The physiological roles of chicken LHRH-I and -II (cLHRH-I and -II) in the regulation of gonadotrophin release were investigated in the domestic chicken. Measurements of the neuropeptides, using specific radioimmunoassays, in brain sections cut in three planes or in grossly dissected brain areas, showed that cLHRH-II occurs in low amounts throughout the brain whereas cLHRH-I is most abundant in the diencephalon. Within the diencephalon, the largest amount of cLHRH-I occurred in the median eminence of the hypothalamus. The amount of cLHRH-I in the median eminence was higher (P less than 0.05) in laying than in out-of-lay hens. No cLHRH-II was detected in the median eminence in either reproductive state. The amount of cLHRH-I in the hypothalamus was increased (P less than 0.05) in cockerels at the onset of puberty and in somatically immature birds after castration. There were no correlated changes in the amounts of hypothalamic cLHRH-II measured in the same experimental samples. Active immunization of laying hens against cLHRH-I but not against cLHRH-II resulted in the complete regression of the reproductive system and a depression in the concentration of plasma LH. These observations, taken together, suggest that gonadotrophin secretion in the hen is more likely to be directly regulated by cLHRH-I than by cLHRH-II.
Assuntos
Galinhas/fisiologia , Gonadotropina Coriônica/metabolismo , Hormônio Liberador de Gonadotropina/análogos & derivados , Hipotálamo/análise , Animais , Química Encefálica , Feminino , Hormônio Liberador de Gonadotropina/análise , Hormônio Liberador de Gonadotropina/imunologia , Hormônio Liberador de Gonadotropina/fisiologia , Imunização , Hormônio Luteinizante/sangue , Ácido Pirrolidonocarboxílico/análogos & derivados , Radioimunoensaio/métodosRESUMO
Galanin gene expression in the anterior pituitary is potently stimulated by estrogen in adult rats. To evaluate the influence of estrogen on galanin during the peripubertal period 30- to 32-day-old female rats were treated with pregnant mare serum gonadotropin (PMSG, 10 IU s.c., 10.00 h). Galanin-like immunoreactivity (galanin-LI) in hypothalamic and pituitary tissues was evaluated 1, 2 or 3 days after PMSG treatment between 17.00 and 19.00 h. The PMSG treatment stimulated 17 beta-estradiol secretion, which induced a midafternoon LH surge 2 days after the PMSG treatment. Concentrations of galanin-LI at the time of this LH surge were elevated 82% in the anterior pituitary and 58% in the hypothalamus (without the median eminence) when compared to saline-treated female rats. On the 3rd day after the PMSG injection, galanin-LI was increased 236% in the anterior pituitary, 88% in the neurointermediate lobe and 39% in the median eminence compared to saline-treated female rats. These changes in galanin-LI were not observed in similarly aged male rats or ovariectomized rats treated with PMSG. In adult male rats, daily injections with 17 beta-estradiol valerate (10 micrograms/daily s.c.) for 1 week increased galanin-LI in the median eminence and neurointermediate lobe to an extent similar to that seen in juvenile female rats following PMSG treatment. In contrast, the high serum levels of 17 beta-estradiol achieved after 17 beta-estradiol valerate treatment increased galanin-LI in the anterior pituitary 65-fold. These studies indicate that galanin-LI is influenced by estrogen in peripubertal and adult rats.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Estradiol/análogos & derivados , Gonadotropinas Equinas/farmacologia , Hipotálamo/análise , Eminência Mediana/análise , Neuropeptídeos/análise , Peptídeos/análise , Hipófise/análise , Maturidade Sexual/efeitos dos fármacos , Animais , Estradiol/farmacologia , Feminino , Galanina , Hipotálamo/efeitos dos fármacos , Masculino , Eminência Mediana/efeitos dos fármacos , Hipófise/efeitos dos fármacos , Ratos , Ratos Endogâmicos , Maturidade Sexual/fisiologiaRESUMO
To investigate possible sex differences in the feedback regulation of growth hormone (GH) secretion, concentrations of immunoreactive GH-releasing hormone (GRF) and somatostatin (SS) were measured in the median eminence (ME) and the hypothalamus of male and female rats bearing the MtTW15 tumor, which secretes high amounts of GH and prolactin (PRL). Four weeks after tumor implantation in male rats, the GRF concentration in the whole hypothalamus, including the ME, was decreased by 37% (0.29 +/- 0.02 vs. 0.46 +/- 0.02 ng/mg protein in intact male controls; p less than 0.001) and the concentration of SS was increased by 40% (11.5 +/- 0.7 vs. 8.1 +/- 0.3 ng/mg protein in male controls; p less than 0.01). In female rats, the presence of tumor for 4 weeks caused a smaller (18%) reduction in GRF concentrations (0.27 +/- 0.02 vs. 0.33 +/- 0.03 ng/mg protein in intact female controls; p less than 0.05) and no significant change in SS concentrations (10.2 +/- 0.08 vs. 9.7 +/- 0.8 ng/mg protein in female controls). Tumor-related changes in GRF and SS concentrations were also more pronounced in male rats than in females, when determined separately in the microdissected ME and in the remaining hypothalamus. These differences occurred despite similar increases in serum GH, PRL and insulin-like growth factor I concentrations in male and female tumor-bearing rats. To assess which hormone (GH or PRL) was responsible for these changes, intact male rats were treated for 10 days with 2 daily s.c. injections of rat GH (rGH; 100 and 250 micrograms/day), rat PRL (100 and 250 micrograms/day) or vehicle.(ABSTRACT TRUNCATED AT 250 WORDS)