The origin and ultrastructural characteristics of corticotropin-releasing factor (CRF)-immunoreactive nerve fibers in the posterior pituitary of the rat

Summary A fine network of corticotropin-releasing factor (CRF)-immunopositive fibers was found in the posterior lobe of the pituitary of the rat. The intermediate and distal lobes were free of CRF-immunoreactivity. Varicose, terminal-like axons were frequently observed around capillary vessels. Surgical isolation of the paraventricular nuclei resulted in a complete disappearance of CRF-immunoreactive fibers from the posterior lobe. CRF-immunopositive fibers show the general characteristics of peptidergic axons. These ultrastructural observations support the idea that CRF is secreted into capillary vessels.     

Ontogenetic development of corticotropin-releasing factor (CRF)-containing neural elements in the brain of the chicken during incubation and after hatching

Summary In chicken embryos of different ages and in young chickens after hatching, neural elements reacting with antibodies generated against synthetic ovine corticotropin-releasing factor (CRF) were studied by means of the peroxidase-anti-peroxidase (PAP) technique at the lightmicroscopic level. CRF-immunoreactivity was first observed in perikarya located in the periventricular part of the hypothalamus on the 14th day of the incubation period. CRF-containing neural elements were detected on the same day of incubation in the external zone of the median eminence, but not in all investigated animals. In extrahypothalamic sites, immunoreactive perikarya were demonstrable in the central gray of the mesencephalon on the 15th day of incubation. Furthermore, immunoreactive cells appeared in other brain regions such as nucleus accumbens and dorsomedial nucleus of the thalamus after hatching. The present observations provide information regarding the functional development of the hypothalamo-hypophyseal-adrenal axis in the chick embryo.     

Neurosecretion in the external and internal zone of the median eminence of the cat and dog

Summary The immunoglobulin-enzyme bridge technique, in association with rabbit antiporcine neurophysin-II has been applied to the median eminence of the dog and cat in order to study the distribution of neurophysin-like antigens throughout this area of the brain and correlate the findings with the corresponding distribution of neurosecretory material (NSM) as revealed by the crotonaldehyde fuchsin stain.Neurophysin and NSM were both present in the hypothalamo-supraoptico-neurohypophysial system—the pathway taken by oxytocin, vasopressin and neurophysin from the hypothalamus to the posterior pituitary lobe.Whereas the tuberoinfundibular tract of the median eminence was almost devoid of NSM, the presence of neurophysin-like material was clearly evident using immunoperoxidase histochemistry. The significance of a protein in the external median eminence possessing determinants cross-reactive against anti-neurophysin serum is discussed.This work was financed by a grant from the Medical Research Council of New Zealand.     

Effect of CRF injected into the median eminence on GH secretion in female rats under different steroid status

To evaluate whether the median eminence (ME) is a site of action of CRF (corticotropin releasing factor) on GH secretion and to determine the possible role of estradiol and progesterone in modifying theses secretion, we injected CRF (0.25, 0.75, 1, and 1.5 nmol of peptide dissolved in 1 μl of water) directly into the ME in three experimental groups of rats: Long-term ovariectomized (OVX); OVX primed by estradiol (OVX±E) and OVX primed by estradiol plus progesterone (OVX±EP). Blood was collected to determine GH (30, 60, 90, and 120 min postinjection) Serum T₃, T₄, and glucose levels were measured in OVX±E rats 30 min postinjection. CRF at all doses studied significantly decreased serum GH levels in the three experimental groups. Serum T₃, T₄, and glucose levels were unchanged after CRF administration. The results suggest that: CRF inhibits “per se” GH secretion, at least in part, by a central action in the ME. The inhibitory effect of CRF on GH is independent of the estrogen/progesterone status of the animal. CRF at ME levels may participate in a variety of stress-related responses, including growth inhibition, through GH suppression.     

Adrenal modulation of the inhibitory effect of corticotropin releasing factor on feeding

Corticotropin releasing factor (CRF) reduces food intake in rats after central administration. In these studies we examined whether the adrenal gland and the vagus were involved in CRF suppression of intake. One hour intake was reduced by a 5 μg (ICV) injection of CRF in sham but not adrenalectomized rats maintained on 0.9% NaCl. In a separate experiment on rats maintained on tap water, the inhibitory effect of CRF (5 μg) lasted at least 4 hours in sham rats whereas adrenalectomized rats did not significantly differ from controls. These experiments suggest that the adrenal gland modulates the feeding response to CRF. As replacement with corticosterone (0.75 mg/kg) in total adrenalectomized rats did not restore responsiveness to 5 or 10 μg of CRF, we next studied whether the adrenal medulla was responsible for the decreased responsiveness to CRF. In rats lacking the adrenal medulla only, food intake was reduced by a 5 μg injection of CRF; in sham rats, intake was significantly reduced by doses as low as 0.1 μg of CRF. An additional experiment examined the effect of gastric vagotomy on the CRF feeding response. Vagotomized rats were as responsive to 5 and 10 μg injections of CRF as sham rats, which suggests that the effect is not dependent on the vagus nerve. These findings indicate that the adrenal gland, primarily the medulla, plays an intermediate role in the reduction of food intake caused by central injections of CRF. This conclusion is consistent with the known effect of CRF on adrenomedullary discharge.     

Effects of synthetic ovine CRF on ACTH, cortisol and blood pressure in sheep

Intravenously administered synthetic ovine CRF at doses of 0.1, 1.0 and 10.0 micrograms/kg increased plasma ACTH and cortisol concentrations in a dose-dependent fashion in unanesthetized sheep. In two unanesthetized sheep, aortic blood pressure remained relatively unaffected after the intravenous administration of CRF at 5 and 20 micrograms/kg. These results suggest that peripherally administered ovine synthetic CRF specifically stimulates the sheep pituitary-adrenal axis. Unlike other species receiving intravenous synthetic ovine CRF, sheep did not show hypotensive effects.     

Production of antiserum to CRF associated with adrenal atrophy in a rabbit

Corticotropin releasing factor (CRF) was recently isolated from ovine hypothalami by its ability to stimulate adrenocorticotropin (ACTH) and β-endorphin release from dispersed rat pituitary cells. Intramuscular injection of synthetic ovine CRF conugated to bovine thyroglobulin with 1-ethyl-3(3-dimethylaminopropyl) carbodiimide and emulsified with Freund's complete adjuvant into a random bred New Zealand white rabbit resulted in antiserum production to CRF associated with adrenal atrophy. A decrease in the level of plasma coticosteroids was associated with an increase in mean total binding of ¹²⁵I-N-Tyr-CRF. Upon sacrifice, a decrease in pituitary content of ACTH and a decrease in adrenal weight and content of corticosteroids was observed in the rabbit producing antiserum to CRF. Adrenal atrophy was histologically verified with an observed decrease in the adrenal cortical zone not reflected in the zona glomerulosa. Individual cells were relatively larger either with more abundant pale cytoplasm or with distinctly vacuolated cytoplasm. The results presented here are consistent with a physiologically necessary role for this CRF or peptides with similar structures in the hypothalamic-pituitary-adrenal axis.     

SPSSM8: An accurate approach for predicting eight-state secondary structures of proteins

Protein eight-state secondary structure prediction is challenging, but is necessary to determine protein structure and function. Here, we report the development of a novel approach, SPSSM8, to predict eight-state secondary structures of proteins accurately from sequences based on the structural position-specific scoring matrix (SPSSM). The SPSSM has been successfully utilized to predict three-state secondary structures. Now we employ an eight-state SPSSM as a feature that is obtained from sequence structure alignment against a large database of 9 million sequences with putative structural information. The SPSSM8 uses a low sequence identity dataset (9062 entries) as a training set and conditional random field for the classification algorithm. The SPSSM8 achieved an average eight-state secondary structure accuracy (Q8) of 71.7% (Q3, 81.6%) for an independent testing set (463 entries), which had an improved accuracy of 10.1% and 4.6% compared with SSPro8 and CNF, respectively, and significantly improved the accuracy of eight-state secondary structure prediction. For CASP 9 dataset (92 entries) the SPSSM8 achieved a Q8 accuracy of 80.1% (Q3, 83.0%). The SPSSM8 was confirmed as an outstanding predictor for eight-state secondary structures of proteins. SPSSM8 is freely available at http://cal.tongji.edu.cn/SPSSM8.     

Effects of dopaminergic system activation on feeding behavior and growth performance of the sea bass (Dicentrarchus labrax): A self-feeding approach

Dopamine is synthesized from l-dopa and subsequently processed into norepinephrine and epinephrine. Any excess neurotransmitter can be taken up again by the neurons to be broken down enzymatically into DOPAC. The effect of dopamine on mammalian food intake is controversial. Mice unable to synthesize central dopamine die of starvation. However, studies have also shown that central injection of dopamine inhibits food intake. The effect of dopaminergic system in the fish feeding behavior has been scarcely explored. We report that the inclusion of l-dopa in the diets results in the activation of sea bass central dopaminergic system but also in the significant increase of the hypothalamic serotonin levels. Dietary l-dopa induces a decrease of food intake and feed conversion efficiency that drives a decline of all growth parameters tested. No behavioral effects were observed after l-dopa treatment. l-dopa treatment stimulated central expression of NPY and CRF. It suggests that CRF might mediate l-dopa effects on food intake but also that CRF neurons lie downstream of NPY neurons in the hierarchical forebrain system, thus controlling energy balance. Unexpectedly, dietary administration of haloperidol, a D₂-receptor antagonist, cannot block dopamine effects but also induces a decline of the food intake. This decrease seems to be a side effect of haloperidol treatment since fish exhibited a decreased locomotor activity. We conclude that oral l-dopa inhibits sea bass food intake and growth. Mechanism could also involve an increase of hypothalamic serotoninergic tone.