Thyrotropin-releasing hormone precursor: characterization in rat brain

To characterize the precursor of mammalian thyrotropin-releasing hormone (TRH), a rat hypothalamic lambda gt11 library was screened with an antiserum directed against a synthetic peptide representing a portion of the rat TRH prohormone. The nucleotide sequence of the immunopositive complementary DNA encoded a protein with a molecular weight of 29,247. This protein contained five copies of the sequence Gln-His-Pro-Gly flanked by paired basic amino acids and could therefore generate five TRH molecules. In addition, potential cleavage sites in the TRH precursor could produce other non-TRH peptides, which may be secreted. In situ hybridization to rat brain sections demonstrated that the pre-proTRH complementary DNA detected neurons concentrated in the parvocellular division of the paraventricular nucleus, the same location as cells detected by immunohistochemistry. These findings indicate that mammalian TRH arises by posttranslational processing of a larger precursor protein. The ability of the TRH prohormone to generate multiple copies of the bioactive peptide may be an important mechanism in the amplification of hormone production.     

Thyrotropin-releasing hormone: regional distribution in rat brain

A sensitive and specific radioimmnunoassay has been used to measure the distribution of thyrotropin-releasing hormone (TRH) in rat brain. All areas of brain tested, except cerebellum, contained readily measurable amounts of TRH. The hypothalamus contained only 31.2 percent of the total brain content of TRH. These results support recent suggestions of central actions for TRH in addition to its hypophysiotropic functions.     

Thyrotropin-releasing hormone in specific nuclei of rat brain

The regional distribution of thyrotropin-releasing hormone (TRH) in rat brain was studied. The greatest concentration of TRH was found in the median eminence. High concentrations were also found in several hypothalamic nuclei. Outside the hypothalamus, relatively large amounts of TRH were found in the septal and preoptic areas.     

Thyrotropin-releasing hormone induces rhythmic bursting in neurons of the nucleus tractus solitarius

The nucleus tractus solitarius (NTS) contains neurons that are part of the central neuronal network controlling rhythmic breathing movements in mammals. Nerve terminals within the NTS show immunoreactivity to thyrotropin-releasing hormone (TRH), a neuropeptide that has potent stimulatory effects on respiration. By means of a brainstem slice preparation in vitro, TRH induced rhythmic bursting in neurons in the respiratory division of the NTS. The frequency of bursting was voltage-dependent and could be reset by short depolarizing current pulses. In the presence of tetrodotoxin, TRH produced rhythmic oscillations in membrane potential whose frequency was also voltage-dependent. These observations suggest that TRH modulates the membrane excitability of NTS neurons and allows them to express endogenous bursting activity.     

Thyrotropin-releasing hormone: biosynthesis by rat hypothalamic fragments in vitro

Biosynthesis of thyrotropin-releasing hormone (L-pyroglutamyl-L-histidyl-L-proline amide) in vitro was studied. Rat hypothalamic fragments were incubated in Krebs-Ringer bicarbonate buffer that contained either (14)C-labeled proline, histidine, or glutamic acid (the three probable precursor amino acids,) and for control purposes each of 16 other naturally occurring amino acids. A number of labeled peptides were synthesized. With the use of synthetic thyrotropin-releasing hormone, detected by the Pauly reagent or with (125)1-labeled thyrotropin-releasing hormone as a marker, thin-layer chromatograms, paper electrophoresis, and carboxymethyl cellulose ion exchange chromatography revealed that only proline, histidine, and glutamic acid were consistently incorporated into peptides associated with the thyrotropin-releasing hormone region. This synthesizing activity was found in stalk median eminence, ventral hypothalamus. and dorsal hypothalamus but not in neural lobe or cerebral cortex. Because the biosynthetic peptide has identical properties with L-pyroglutamyl-L-histidyl-L-proline amide, it is probable that rat thyrotropin-releasing hormone is similar or identical to both bovine and porcine thyrotropin-releasing hormone and that the native material is present in the pyroglutamyl form in tissues.     

Effects of cold stress on hypothalamic corticotrophin-releasing and thyrotropin-releasing hormone messenger RNA levels in chickens

There is increasing evidence that stress can activate the hypothalamic-pituitary-adrenal-axis and hypothalamic-pituitary- thyroid-axis,and further affect the synthesis and secretion of corticotrophin-releasing hormone(CRH)and thyrotropin-releasing hormone(TRH).To evaluate the effect of cold stress on the hypothalamic CRH and TRH messenger RNA(mRNA)levels in Yisha chickens,male Yisha chickens were subjected to acute(1,6,12 h)and chronic(5,10,20 d)cold stress(12±1)℃.Hypothalami were collected for assessment of mRNA levels by semi-quantitative RT-PCR.Acute stress resulted in a significant decrease of CRH mRNA levels at 6 and 12 h,and a significant increase of TRH mRNA levels at every stress time point.Chronic cold stress resulted in a significant increase of CRH mRNA levels and a significant decrease of TRH mRNA levels compared with the control group at every stress time point.The results suggest that the two genes differently respond to cold stress at the mRNA levels.And the different degrees of cold stress will produce different effects on the identical gene.     

Human lactational and ovarian response to endogenous prolactin release

Radioimmunoassayable prolactin rises in postpartum women during nursing and after intravenous thyrotropin-releasing hormone (TRH). Prolactin release induced by TRH can be dissociated from the postsuckling response. In addition to this, increases in endogenous prolactin secretion are followed by marked breast engorgement and milk letdown, especially after intravenous TRH. In this group of breast-feeding women, vaginal smears remained atrophic even up to 410 postpartum days. Prolactin appears to influence the production of breast milk, and the maintenance of a regular nursing pattern seems to promote the maintenance of ovarian unresponsiveness to circulating gonadotropins.     

Kisspeptin在青春期雌性皖西白鹅下丘脑和垂体的定位

运用免疫组化PV-9000二步法和DAB显色技术,研究Kisspeptin在青春期雌性皖西白鹅下丘脑和垂体的定位.免疫组化DAB显色结果表明,在下丘脑后内侧核、室周弓状核和室旁核均可观察到大量Kisspeptin免疫反应阳性神经元,下丘脑室周区及外侧区也可观察到不同程度的Kisspeptin阳性细胞;腺垂体中亦可见少量免疫阳性产物,但神经垂体无阳性细胞.Kisspeptin在雌性青春期皖西白鹅下丘脑和垂体中均有分布,初步揭示Kisspeptin在调节促性腺激素的分泌和参与生殖调控以及青春期的启动中均具有非常重要的作用.     

鸵鸟雏鸟下丘脑室旁核发育的组织学及GABA的表达

【目的】观察鸵鸟雏鸟下丘脑室旁核（paraventricular hypothalamic nucleus,PVN）的显微结构和细胞凋亡的发育特点,探明γ-氨基丁酸（γ-aminobutyric,GABA）阳性细胞在PVN内的年龄增长性变化。【方法】以1、45和90 d健康非洲鸵鸟雏鸟为实验动物,利用H.E染色技术、原位缺口末端标记（TUNEL）技术和免疫组化定位表达（SABC）技术,研究鸵鸟雏鸟下丘脑PVN的发育特点。【结果】①鸵鸟雏鸟下丘脑PVN的组织学结构随日龄的增长而逐渐发育成熟,其中PVN小细胞占细胞总数的比例在45 d时最低,明显高于1和90 d的小细胞核;②鸵鸟雏鸟下丘脑PVN内存在细胞凋亡现象,凋亡呈年龄增长性减少;③鸵鸟雏鸟下丘脑PVN内可检测到GABA阳性产物,表达量呈年龄增长性增加的趋势,其中 PVN大细胞部（mPVN）和小细胞部（pPVN）的平均积分光密度均在45 d时最高,显著高于1和90 d的（P＜0.01）。【结论】随着日龄的增长,鸵鸟雏鸟下丘脑PVN的结构和功能不断完善,其在出生后早期（1—45 d）发育较快。     

Inhibition of gastric acid secretion in rats by intracerebral injection of corticotropin-releasing factor

Intracisternal injection of ovine corticotropin-releasing factor (CRF) into the pylorus-ligated rat or the rat with gastric fistula resulted in a dose-dependent inhibition of gastric secretion stimulated with pentagastrin or thyrotropin-releasing hormone. When injected into the lateral hypothalamus--but not when injected into the cerebral cortex--CRF suppressed pentagastrin-stimulated acid secretion. The inhibitory effect of CRF was blocked by vagotomy and adrenalectomy but not by hypophysectomy or naloxone treatment. These results indicate that CRF acts within the brain to inhibit gastric acid secretion through vagal and adrenal mechanisms and not through hypophysiotropic effects.     

雌二醇对大鼠下丘脑神经激肽B表达的影响

为研究雌二醇(Estradiol,E2)对大鼠下丘脑神经激肽B(Neurokinin B,NKB)表达的影响,采用免疫组化PV-9003二步法以及DAB显色技术,对外源性E2处理组(E2组)、花生油处理组(C组)、卵巢摘除+E2组(OVX+E2组)和卵巢摘除组(OVX组)大鼠下丘脑的NKB进行检测。结果表明:大鼠阴门开启时间E2组(29.00±0.707)与OVX+E2组(30.20±0.084)早于对照组(43.60±2.074)(P0.01)。NKB主要分布于所有大鼠下丘脑的弓状核、腹内侧核和室旁核,且OVX组和C组室周核上也有少量表达(P0.05),但各组间NKB的平均光密度值有差异,OVX组大鼠下丘脑弓状核(0.33±0.49)、腹内侧核(0.31±0.54)和室旁核(0.30±0.55)均显著高于OVX+E2、C和E2组(P0.05)。雌激素抑制大鼠下丘脑NKB的表达,NKB可能通过雌激素调节生殖作用。     

Paradoxical elevation of growth hormone by intraventricular somatostatin: possible ultrashort-loop feedback

Somatostatin, the growth hormone-inhibiting factor, when microinjected into the third ventricle of the rat brain, paradoxically induced the release of growth hormone. A pituitary site of action having been ruled out, this result supports the concept that exogenous somatostatin within the hypothalamus acts either to suppress the release of somatostatin from somatostatin-containing neurons, possibly via an ultrashort-loop feedback mechanism, or to augment release of hypothalamic growth hormone-releasing factor, thereby inducing a release of growth hormone. Injection of somatostatin into the third ventricle also decreased plasma concentrations of luteinizing hormone, follicle-stimulating hormone, and thyroid-stimulating hormone, probably by inhibiting the release of luteinizing hormone-releasing factor and thyrotropin-releasing factor.     

Cyclic adenosine monophosphate: selective increase in caudate nucleus after administration of L-dopa

Treatment with the dopamine precursor L-dopa produced a significant accumulation of adenosine 3',5'-monophosphate (cyclic AMP) in the caudate nucleus of the rat. In contrast, there was no change in the amount of cyclic AMP in the cerebellum. Accumulation of cyclic AMP in the caudate nucleus after administration of L-dopa was prevented by prior treatment with the decarboxylase inhibitor RO 4-4602. These observations and those in other laboratories support the assumption that dopamine formed from L-dopa selectively activates striatal adenylate cyclase. The in vivo activation of adenylate cyclase after treatment with L-dopa may be a useful model for studying neurological and psychiatric disorders that are thought to involve the dopaminergic system of the brain.     

Narcotic drugs: effects on the serotonin biosynthetic systems of the brain

The effects of short- and long-term administration of morphine on the activity of two measurable forms of rat brain tryptophan hydroxylase were studied. Morphine administration produced an immediate decrease and a longterm increase in the nerve ending (particulate) enzyme activity but did not change the cell body (soluble) enzyme activity. Cocaine administration demnonstrated a short-term decrcease in measurable nerve eniding enzyme activity that was due to the inhibition of the high affinity uptake (the Michaelis constant, K(m) is 10-(5) molar) of trytophan, the serotonin precursor. Cocaine did not aflect the low affinity uptake K(m) = 10-(5) molar) of tryptophan. Both the uptake of the precursor and the enizymiie activity appeared to be drug-sensitive regullatory processes in the biosynthlesis of serotonin.     

Immunoreactive dynorphin-(1-8) and corticotropin- releasing factor in subpopulation of hypothalamic neurons

Immunoreactive corticotropin-releasing factor (CRF) and dynorphin-(I-8) were visualized in rat hypothalamus by immunohistofluorescence with specific antibodies. In brains from colchicine-treated, adrenalectomized rats, neuronal perikarya with immunoreactive CRF were observed in the paraventricular nucleus of the hypothalamus. The CRF occurred together with the dynorphin-(1-8). However, the CRF immunoreactivity occurred only in a subpopulation of the dynorphin-(1-8) immunoreactive cells. These findings suggest that there may be a functional interrelationship of CRF with dynorphin-related opioid peptides and provide further evidence that neurons may contain more than one bioactive substance.     

Choline: high-affinity uptake by rat brain synaptosomes

Synaptosomes from rat brain accumulate choline by two kinetically distinct processes, a high-affinity uptake system [Michaelis constant (K(m)) = 1 x 10(-6)M], and a low-affinity system (K(m) = 9 x 10(-5)M). The high-affinity uptake system requires sodium, and is associated with considerable formation of acetylcholine. The low-affinity uptake system is less dependent on sodium, and does not appear to be associated with a marked degree of acetylcholine formation. The high-affinity choline uptake appears to represent selective choline accumulation by cholinergic neurons.     

Cholecystokinin receptors in the brain: characterization and distribution

Specific cholecystokinin binding sites in particulate fractions of rat brain were measured with iodine 125-labeled Bolton-Hunter cholecystokinin, a cholecystokinin analog that has full biological activity. Binding was detected in brain regions known to contain immunoreactive cholecystokinin. Binding was saturable, reversible, of high affinity (dissociation constant, 1.7 x 10(-9) M), and was inhibited by cholecystokinin analogs but not by unrelated hormones.     

Release of multiple hormones by a direct action of interleukin-1 on pituitary cells

Exposure to bacterial endotoxins has long been known to stimulate the release of anterior pituitary hormones; administration of endotoxin was at one time a common clinical test of anterior pituitary function. Endotoxin is a potent stimulus for production of the endogenous pyrogenic protein, interleukin-1 (IL-1), by macrophages and monocytes. The possibility that IL-1 has a direct effect on the secretion of hormones by rat pituitary cells in a monolayer culture was investigated. Recombinant human IL-1 beta stimulated the secretion of adrenocorticotropic hormone, luteinizing hormone, growth hormone, and thyroid-stimulating hormone. Increased hormone secretion into culture supernatants was found with IL-1 concentrations ranging from 10(-9) M to 10(-12) M. Prolactin secretion by the monolayers was inhibited by similar doses. These concentrations of IL-1 are within the range reported for IL-1 in serum, suggesting that IL-1 generated peripherally by mononuclear immune cells may act directly on anterior pituitary cells to modulate hormone secretion in vivo. Incubation of IL-1 solutions with antibody to IL-1 neutralized these actions. These pituitary effects of IL-1 suggest that this monokine may be an important regulator of the metabolic adaptations to infectious stressors.