Injection Of Corticotropin-releasing Hormone Research Articles

Central administration of growth hormone-releasing hormone and corticotropin-releasing hormone stimulate downstream movement and thyroxine secretion in fall-smolting coho salmon ( Oncorhynchus kisutch)

The ultimate signal triggering downstream migration in anadromous salmonids is unknown. A plasma surge of T 4 (T 4 surge) occurs during downstream migration in salmonids; however, the causal relationship between migratory behavior and the T 4 surge is not well known. We first examined the progression of smolt indicators (skin silvering, condition factor (CF), gill Na +, K +-ATPase (NKA) activity and plasma T 4 levels) in underyearling, fall-smolting coho salmon ( Oncorhynchus kisutch) from August to December. In November, the fish showed the characteristics of fully developed smolts, i.e. the skin completely covered with silvery scales, CF at a nadir, and peak NKA activity and plasma T 4 levels. Based on these results, we examined the effects of four neuropeptides, thyrotropin-releasing hormone (TRH), corticotropin-releasing hormone (CRH), growth hormone-releasing hormone (GHRH), and gonadotropin-releasing hormone (GnRH), on the downstream movement (negative rheotaxis) and T 4 surge in fully smoltified underyearling coho salmon. The experiment was run in circular-shaped channel tanks and the neuropeptide treatment was performed as intracerebroventricular (ICV) injections. ICV injection of GHRH and CRH stimulated both downstream movement and plasma T 4 level. TRH injection stimulated plasma T 4 level but suppressed downstream movement. GnRH injection had no effect. It is hypothesized that GHRH and CRH play key roles in triggering downstream migration of anadromous salmonids, and that the accompanying T 4 surge is a consequence of the neuroendocrine processes that trigger migration.

Central Deficiency of Corticotropin-Releasing Hormone Receptor Type 1 (CRH-R1) Abolishes Effects of CRH on NREM But Not on REM Sleep in Mice

Corticotropin-releasing hormone (CRH) is the major activator of the hypothalamic-pituitary-adrenocortical (HPA) system and orchestrates the neuroendocrine, autonomous as well as behavioral responses to stress. Many studies suggest an influence of CRH on sleep-wake regulation even in the absence of stressors. However, none of these studies yet clearly distinguished between central and peripheral effects of CRH. Therefore, we investigated in CNS-specific CRH receptor type 1 deficient mice whether centrally administered CRH could induce its sleep-wake modulatory effects without peripheral induction of HPA activity. Male mice (C57BL/6J, CNS-specific CRH-R1 knockout [CKO] mice and their control littermates [CL]) were intracerebroventricularily (i.c.v.) injected with vehicle or 3 different doses of CRH shortly before the beginning of the light period. Electroencephalogram (EEG) and electromyogram (EMG) were monitored to compare the effects of CRH on vigilance states with or without presence of central CRH-R1. To quantify HPA-axis reactivity to CRH injections in CKO and CL animals, blood samples were analyzed to determine plasma corticosterone concentrations. I.c.v. injections of CRH promoted wakefulness while decreasing NREMS in C57BL/6J and CRH-R1 CL animals, whereas such changes were not exerted in CKO mice. However, REMS suppression after CRH application persisted in all animals. I.c.v. injected CRH increased plasma corticosterone levels in both CL and CKO mice. The results demonstrated that CRH has a major impact on wake and NREMS regulation which is predominantly mediated through central CRH-R1. Peripheral actions of CRH, i.e., elevated HPA activity, may interfere with its central effects on REMS but not on NREMS suppression.

CRH Signaling

There is an urgent need to generate new drugs or improve existing ones in the pharmacology of mood disorders. The corticotropin-releasing hormone (CRH) system is closely involved in the development and course of depression, and drugs targeting this system arguably offer hope to improve the current tools for drug treatment of depression. Recent clinical studies in depressed patients showed that CRHR1 antagonists improve clinical symptoms of anxiety and depression and reduce stress hormone release following psychosocial stress. These effects of CRHR1 antagonists were not associated with reduced secretory capacity of corticotrophic cells because of CRH receptor abundance at the pituitary level, which contrasts with CRH receptors in the brain. This is in accordance with previous studies showing that CRH injections into the mouse brain activate MAPK pathways in a brain region-specific manner pointing toward differences in signaling pathways beyond the receptor level. We will highlight this and discuss how these brain area-specific differences may offer opportunities for drug discovery. An additional puzzle in the search of new targets for depression is the lack of bona fide animal models helping to discover the antidepressants that are not monoamine based. We recently developed a conditional mouse model that overexpresses CRH in a spatio-temporal-regulated fashion and permits to dissect precisely the contribution of different brain areas to the CRH-dependent behaviors. Recent findings obtained with this mouse model and its usefulness in the context of the CRH-dependent, region-specific changes in depression will be discussed.

Covariance Between Psychological and Endocrine Responses to Pharmacological Challenge and Psychosocial Stress: A Question of Timing

To test if the covariance of hypothalamus-pituitary-adrenal (HPA) axis and subjective-psychological responses to stress is dependent on different dynamics of these systems. Although stress theories typically assume substantial correlations of psychological and endocrine stress responses, studies have produced inconsistent results. One reason for this might be imperfect coupling of the different stress response systems. However, inconsistent correlations might also be a result of different on-/offsets of these stress responses, i.e., specific dynamics of the systems. HPA axis indicators and subjective-psychological states were repeatedly and synchronously measured in a pharmacological challenge test (injection of corticotropin-releasing hormone and infusion of arginine vasopressin; Study 1; n = 42) and a psychosocial stress situation (Trier Social Stress Test; Study 2; n = 219). Cross-correlation analysis was used to test for lag effects in HPA axis reactivity and psychoendocrine responses. Analyses revealed high cross-correlations of adrenocorticotropic hormone with cortisol responses (up to r = .80 in Study 1 and r = .56 in Study 2) and positive associations of psychological with endocrine stress responses (up to r = .48 in Study 1 and r = .54 in Study 2) at nonzero lags. Subjective-psychological responses preceded HPA axis responses. Moreover, high levels of cortisol were associated with lower later levels of anxiety and activation. The findings suggest that psychoendocrine stress responses are more closely coupled than previous studies suggested. Due to different dynamics of the systems, endocrine responses lag behind psychological responses.

Corticotropin-releasing hormone receptors in the medial prefrontal cortex regulate hypothalamic–pituitary–adrenal activity and anxiety-related behavior regardless of prior stress experience

The hypothalamic–pituitary–adrenal (HPA) axis habituates, or gradually decreases its activity, with repeated exposure to the same stressor. During habituation, the HPA axis likely requires input from cortical and limbic regions involved in the processing of cognitive information that is important in coping to stress. Brain regions such as the medial prefrontal cortex (mPFC) are recognized as important in mediating these processes. The mPFC modulates stress-related behavior and some evidence suggests that the mPFC regulates acute and repeated stress-induced HPA responses. Interestingly, corticotropin-releasing hormone (CRH)-1 receptors, which integrate neuroendocrine, behavioral and autonomic responses to stress, are localized in the mPFC but have not been specifically examined with respect to HPA regulation. We hypothesized that CRH receptor activity in the mPFC contributes to stress-induced regulation of HPA activity and anxiety-related behavior and that CRH release in the mPFC may differentially regulate HPA responses in acutely compared to repeatedly stressed animals. In the present experiments, we found that blockade of CRH receptors in the mPFC with the non-selective receptor antagonist d-Phe-CRH (50 ng or 100 ng) significantly inhibited HPA responses compared to vehicle regardless of whether animals were exposed to a single, acute 30 min restraint or to the eighth 30 min restraint. We also found that intra-mPFC injections of CRH (20 ng) significantly increased anxiety-related behavior in the elevated plus maze in both acutely and repeatedly restrained groups compared to vehicle. Together, these results suggest an excitatory influence of CRH in the mPFC on stress-induced HPA activity and anxiety-related behavior regardless of prior stress experience.

Sex differences in plasma corticosterone release in undisturbed chickens ( Gallus gallus) in response to arginine vasotocin and corticotropin releasing hormone

In birds, two neuropeptides, corticotropin releasing hormone (CRH) and arginine vasotocin (AVT), are major regulators of the hypothalamo-pituitary-adrenal axis (HPA) during the stress response. In birds, however, the relative efficacy of CRH and AVT to stimulate the HPA axis in males and females remains unknown. The purpose of this study was to determine the time course of CORT release following central CRH and AVT administration to male and female chickens. Chickens were fitted with a stainless steel cannula surgically implanted in the lateral ventricle and a catheter chronically inserted in the jugular vein. Birds were housed individually in cages behind a one-way glass partition and unnecessary noise was avoided during the sampling period. Each bird received a single 5.0 μl intracerebroventricular (ICV) injection of either saline (SAL), AVT (10 and 100 pmol), or CRH (10 and 100 pmol). Blood was sampled remotely every 15 min for 2 h and plasma CORT was determined by radioimmunoassay. There was a significant increase in plasma CORT concentration in males injected with 100 pmol AVT beginning at 15 min post-injection through 2 h compared with SAL injected birds. In males, injection of 100 pmol CRH was significantly more effective in releasing CORT compared to an equal molar concentration of AVT or SAL. In females, ICV injection of 100 pmol AVT induced moderate increase in CORT levels. In contrast, 100 pmol CRH significantly increased plasma CORT compared to SAL injected controls but the CORT response was nearly 50% less than that obtained in males.

Physiological and behavioural effects of an intracerebroventricular injection of corticotropin releasing hormone in goats

This study investigated the effects of an intracerebroventricular (ICV) injection of corticotropin releasing hormone (CRH) on physiological and behavioural responses in goats. In Experiment 1, saline (control) or saline plus 25 μg of ovine CRH was injected into the third ventricle of castrated male goats. CRH increased plasma cortisol (Cor) levels markedly within 15 min, but had little effect on plasma glucose (Glu). Compared with saline injected goats, CRH decreased the total duration of lying behaviour but increased its frequency, and suppressed rumination and self-grooming. In Experiment 2, the effects of an intravenous (IV) injection of human adrenocorticotropic hormone (ACTH) (1–24) (0.1 mg) were examined and an IV injection of saline was used as control. ACTH increased plasma Cor levels markedly, but did not change any behaviour compared with controls. It was concluded that CRH mediated the response of the hypothalamus-pituitary-adrenal (HPA) axis and behaviour following stress in goats, although the CRH-induced behavioural changes were independent of the HPA axis and seemed to be the result of direct action within the central nervous system.

Involvement of Intra-articular Corticotropin-releasing Hormone in Postoperative Pain Modulation

Opioid receptors are expressed on peripheral nerve endings and opioid peptides (beta-endorphin, END) are produced in various immune cells of synovial tissue after knee trauma. Because corticotropin-releasing hormone (CRH) acts through its receptors on END-containing immune cells, this randomized controlled trial investigated whether the intra-articular (IA) injection of CRH reduces postoperative pain intensity and supplemental analgesic consumption in patients undergoing arthroscopic knee surgery. Patients were randomly assigned to one of the following IA and IV treatments: group saline (SAL) (n=17) received isotonic SAL IA and 10 microg CRH IV; group CRH (n=16) received 10 microg CRH IA and SAL IV; group CNL (n=18) received 10 microg CRH plus 0.12 mg naloxone IA and SAL IV. Patients pain intensity at rest and during exercise, cortisol plasma concentrations as well as supplemental analgesics were documented. Immunohistochemistry analyzed colocalization of CRH receptors and END. IA but not IV CRH resulted in a significant but short lasting reduction of postoperative pain under both resting and exercise conditions without changes in cortisol plasma concentrations. Coadministration of naloxone reversed this pain reduction under resting but not exercise conditions. The majority of CRH receptor expressing cells contained END within synovial tissue. In conclusion, this first clinical trial provides preliminary evidence for a short but not robust analgesic effect of a single dose of IA CRH in patients undergoing arthroscopic knee surgery. Further clinical studies will have to examine different doses of IA CRH-induced analgesia and to support the involvement of opioid peptides.

Role of corticotropin-releasing hormone in irritable bowel syndrome and intestinal inflammation

Corticotropin-releasing hormone (CRH) is a major mediator of stress response in the brain-gut axis. Irritable bowel syndrome (IBS) is presumed to be a disorder of the brain-gut link associated with exaggerated response to stress. We first showed that peripheral administration of CRH aggravated visceral sensorimotor function as well as adrenocorticotropic hormone (ACTH) response in IBS patients. We then administered alpha-helical CRH (alphahCRH), a non-selective CRH receptor antagonist among IBS patients. Electrical stimulation of the rectum induced significantly higher motility indices of the colon in IBS patients than in the controls. This response was significantly suppressed in IBS patients but not in the controls after administration of alphahCRH. Administration of alphahCRH induced a significant increase in the barostat bag volume of the controls but not in that of IBS patients. alphahCRH significantly reduced the ordinate scale of abdominal pain and anxiety evoked by electrical stimulation in IBS patients. Plasma ACTH and serum cortisol were generally not suppressed by alphahCRH. Last, administration of CRH1-receptor (CRH-R1) specific antagonist blocked colorectal distention-induced sensitization of the visceral perception in rats. Moreover, pretreatment with CRH-R1 antagonist blocked colorectal distention-induced anxiety, which was measured with elevated plus-maze, in rats. Evidence supporting the concept that peripheral CRH and CRH-R1 play important roles in brain-gut sensitization is increasing. Several studies have identified immunoreactive CRH and urocortin as well as CRH-R1 and CRH-R2 mRNAs in human colonic mucosa. In addition, reverse transcription-polymerase chain reaction has revealed the expression of CRH-R1 mRNA in both the myenteric and submucosal plexus in the guinea pig. Application of CRH has been shown to evoke depolarizing responses associated with elevated excitability in both myenteric and submucosal neurons. On the other hand, peripheral injection of CRH has been reported to induce discrete effects on colonic secretory and motor function, and permeability. There are functional differences between CRH-R1 and CRH-R2. For instance, activation of CRH-R1 causes a proinflammatory response, whereas stimulation of CRH-R2 provokes anti-inflammatory changes. In addition, there is evidence of the contrasting roles of CRH-R1 and CRH-R2 in visceral nociception. While CRH-R1 is involved in the pro-nociceptive effects of visceral pain, CRH-R2 mediates an anti-nociceptive response. These findings suggest the major role of CRH in stress-related pathophysiology of IBS and possibly in inflammation of the intestinal mucosa.

Chronic administration of fluoxetine alters locomotor behavior, but does not potentiate the locomotor stimulating effects of CRH in juvenile Chinook salmon (Oncorhynchus tshawytscha)

The present study investigated: 1) the behavioral effects of chronic administration of a serotonin uptake inhibitor (fluoxetine) in juvenile Chinook salmon, Oncorhynchus tshawytscha and, 2) whether chronic administration of fluoxetine alters the behavioral effects of corticotropin-releasing hormone (CRH). Chronic (20 day) treatment with fluoxetine decreased locomotor activity when compared to fish given long-term injections of saline. An intracerebroventricular (i.c.v.) injection of CRH had no effect on locomotor activity following a 20 day intraperitoneal treatment with either saline or fluoxetine. Chronic treatment with fluoxetine also increased the amount of time fish spent near the center of the tank. A similar increase was seen in fish given a chronic intraperitoneal (i.p.) series of saline followed by an acute i.c.v. injection of CRH. However, the effect was not additive when fish were given chronic i.p. injections of fluoxetine followed by an acute i.c.v. injection of CRH. These results provide evidence to support the hypothesis that the serotonergic system is involved in mediating locomotor activity and habitat choice in teleosts.

In vivo effects of corticotropin-releasing hormone on femoral adipose tissue metabolism in women

To investigate whether i.v. injected corticotropin-releasing hormone (CRH) (1 microg/kg) has a direct effect on adipose tissue metabolism in humans. Double-blinded, placebo-controlled, crossover study. Twelve healthy normal weight female volunteers (age 20-37 years, body mass index: 22.75+/-1.33 kg/m(2)) Assessment of local generation of glycerol, and glucose in adipose tissue by microdialysis. Measurement of adipose tissue and skin blood flow by laser Doppler flowmetry. Injection of CRH acutely increases interstitial concentrations of glycerol (19.0+/-5.4%, P<0.05) and glucose (13.5+/-5.8%, P<0.05) reaching peak levels after 15 min. Plasma glycerol increases in parallel (Delta=16.7+/-5.9% after 15 min (P<0.05)), whereas plasma glucose remains unaffected. Changes in tissue blood flow do not explain interstitial metabolite alterations. Initial CRH effects on adipose tissue metabolism are short lasting and disappear after 15 min. The importance of CRH on human energy metabolism is underlined by the present in vivo study demonstrating peptidergic effects on lipolysis and glucose homeostasis in human subcutaneous adipose tissue.

Corticotropin-Releasing Hormone Reduces Pressure Pain Sensitivity in Humans without Involvement of β-Endorphin(1–31), but Does Not Reduce Heat Pain Sensitivity

In the present study the effects of intravenously administered corticotropin-releasing hormone (CRH) on the release of proopiomelanocortin (POMC) derivatives such as adrenocorticotropic hormone (ACTH), β-lipotropin (β-LPH) and β-endorphin (β-END) as well as direct effects of CRH on pain sensitivity were examined. In 16 healthy volunteers we studied the effects of 100 µg intravenously administered CRH in absence or presence of 12 mg naloxone on heat or pressure pain sensitivity, using a double-blind, cross-over and placebo-controlled design. To evaluate analgesic effects of CRH via release of POMC derivatives, we determined plasma concentrations of β-END-immunoreactive material (IRM), authentic β-END (β-END(1–31)) and β-LPH IRM, in parallel with heat and pressure pain tolerance thresholds before and 15 and 30 min after treatment with CRH (or placebo), and 5 min after naloxone (or placebo) administration which was administered 40 min after CRH (or placebo) injection. CRH increased levels of β-END IRM, β-END(1–31) and β-LPH IRM. As compared to β-END IRM levels measured by a commercial RIA kit, the β-END(1–31) levels determined by a highly specific two-site RIA, proved to be remarkably small. Furthermore, CRH did not induce increases of heat pain tolerance thresholds, but of pressure pain tolerance thresholds, which, however, were not reversible by naloxone. Neither β-END nor β-LPH IRM nor β-END(1–31) levels correlated with heat or pressure pain tolerance thresholds. We conclude that CRH does not modulate heat, but pressure pain; POMC derivatives like β-END IRM, β-END(1–31) or β-LPH do not mediate this effect.

Corticotropin-releasing hormone in the lateral parabrachial nucleus inhibits sodium appetite in rats

The present study investigated the role of corticotropin-releasing hormone (CRH) in the lateral parabrachial nucleus (LPBN) in the behavioral control of body fluid homeostasis by determining the effect of bilateral injections of the CRH receptor antagonist, alpha-helical corticotropin-releasing factor (CRF)(9-41), and the CRH receptor agonist, CRH, on sodium chloride (salt appetite) and water (thirst) intake. Groups of adult, male Sprague-Dawley rats had stainless-steel cannulas implanted bilaterally into the LPBN and were sodium depleted or water deprived. Bilateral injections of alpha-helical CRF(9-41) into the LPBN significantly potentiated water and salt intake in the sodium-depleted rats when access to fluids was restored. Bilateral injections of alpha-helical CRF(9-41) into the LPBN (1.0 microg) also increased sodium appetite in water-deprived rats. Conversely, in sodium-depleted animals, bilateral injections of CRH inhibited sodium chloride intake. These results suggest that there is an endogenous CRH inhibitory mechanism operating in the LPBN to modulate the intake of sodium (salt appetite). This mechanism may contribute to the behavioral control of restoration of body fluid homeostasis in sodium-deficient states.

The hypothalamic–pituitary–adrenal (HPA) axis in habitual smokers

Nicotine is a strong activator of the hypothalamus pituitary adrenal (HPA) axis. Smoking of only two cigarettes consistently activates the HPA axis of habitual smokers. However, while being a habitual smoker only induces small changes of basal HPA axis activity, smoking induces an attenuated responsiveness of the HPA axis to psychological stress, but not to injection of corticotropin releasing hormone (CRH) or physiological load. The latter points to alterations at hypothalamic or other central structures. The further consequences of decreased HPA axis responsiveness are discussed. Chronic inflammation of the airways is a common consequence of habitual smoking, and smokers often present with low-grade systemic inflammation, which may be mediated by HPA axis alterations. However, habitual smokers' monocytes are reported to show an increased sensitivity towards the inflammation suppressing effects of cortisol, while on the one hand, inflammation of the airways appears to be relatively resistant towards glucocorticoid treatment. In conclusion, this pattern of attenuated cortisol responses and decreased glucocorticoid sensitivity may be causally related to disinhibition of inflammatory processes and thereby further stimulate adverse health outcomes, such as airway inflammation or atherosclerosis.

Corticotropin-releasing hormone activates ERK1/2 MAPK in specific brain areas

Corticotropin-releasing hormone (CRH) coordinates hormonal and behavioral responses to stress. The mitogen-activated protein kinase extracellular signal-related kinase 1/2 (ERK1/2) mediates several functions in different forebrain structures and recently has been implicated in CRH signaling in cultured cells. To study in vivo CRH-mediated activation of central ERK1/2, we investigated the expression pattern of the phosphorylated ERK1/2(p-ERK1/2) in the mouse brain after intracerebroventricular CRH injections. As shown by immunohistochemistry and confocal microscopy analysis, CRH administration increased p-ERK1/2 levels specifically in the CA3 and CA1 hippocampal subfields and basolateral complex of the amygdala, both structures related to external environmental information processing and behavioral aspects of stress. Other regions such as hypothalamic nuclei and the central nucleus of the amygdala, also related to central CRH system but involved in the processing of the ascending visceral information and neuroendocrine-autonomic response to stress, did not show CRH-mediated ERK1/2 activation. To dissect the involvement of CRH receptor 1 (CRHR1) and CRHR2, we used conditional knockout mice in which Crhr1 is inactivated in the anterior forebrain and limbic structures. The conditional genetic ablation of Crhr1 inhibited the p-ERK1/2 increase, underlining the involvement of CRHR1 in the CRH-mediated activation. These findings underscore the fact that CRH activates p-ERK1/2 through CRHR1 only in selected brain regions, pointing to a specific role of this pathway in mediating behavioral adaptation to stress.

Corticotropin-releasing hormone-mediated metamorphosis in the neotenic axolotl Ambystoma mexicanum: Synergistic involvement of thyroxine and corticoids on brain type II deiodinase

In the present study, morphological changes leading to complete metamorphosis have been induced in the neotenic axolotl Ambystoma mexicanum using a submetamorphic dose of T 4 together with an injection of corticotropin-releasing hormone (CRH). An injection of CRH alone is ineffective in this regard presumably due to a lack of thyrotropic stimulation. Using this low hormone profile for induction of metamorphosis, the deiodinating enzymes D2 and D3 known to be present in amphibians were measured in liver and brain 24 h following an intraperitoneal injection. An injection of T 4 alone did not influence liver nor brain D2 and D3, but dexamethasone (DEX) or CRH alone or in combination with T 4 decreased liver D2 and D3. Brain D2 activity was slightly increased with a higher dose of DEX, though CRH did not have this effect. A profound synergistic effect occurred when T 4 and DEX or CRH were injected together, in the dose range leading to metamorphosis, increasing brain D2 activity more than fivefold. This synergistic effect was not found in the liver. It is concluded that brain T 3 availability may play an important role for the onset of metamorphosis in the neotenic axolotl.

Metabolic effects of intravenous corticotropin releasing hormone on adipose tissue metabolism in humans

Introduction: Apart from its well known function in the central nervous system corticotropin releasing hormone (CRH) seems to have an additional role in peripheral tissues. Besides in the myocardium and skin, CRH receptors (CRH-R) have also been found in human adipose tissue and intravenous administration of CRH in humans leads to an increase of energy expenditure and fatty acid oxidation. In order to further clarify the role of CRH in adipose tissue we investigated changes in adipose tissue metabolites and blood flow after injection of CRH in humans.

A stress-induced anxious state in male rats: Corticotropin-releasing hormone induces persistent changes in associative learning and startle reactivity

Exposure to intense inescapable stressors induces a persistent anxious state in rats. The anxious state is evident as increased sensory reactivity and enhanced associative learning. We examine whether similar neurobehavioral changes are observed after intracerebroventricular (ICV) administration of corticotropin releasing hormone (CRH). Two behaviors were observed: acoustic startle responses (ASRs) and acquisition of the classically conditioned eyeblink response. Male Sprague-Dawley rats were administered ICV CRH either in a single dose (1.0 microg/rat) or in three doses each separated by 30 min. Exaggerated ASRs were evident 2 hours after either CRH treatment; however, only the rats given three injections exhibited a persistently exaggerated ASR apparent 24 hours after CRH treatment. Rats administered three injections of CRH also exhibited faster acquisition of the eyeblink conditioned response beginning 24 hours after treatment. Yet, we did not find evidence for a persistent activation of the HPA-axis response; three CRH injections did not lead to elevated basal plasma corticosterone levels the following morning. Repeated treatment with CRH over a 1.5-hour period models some of the behavioral changes observed after exposure to intense inescapable stressors.

A technique for long-term implantation of a microcatheter into the third ventricle of post-pubertal Chinese Meishan pigs based on ventriculography

The objectives of this study were to implant a microcatheter into the third ventricle of post-pubertal Chinese Meishan pigs, and to maintain the microcatheter for a long time without causing stress. Fourteen pigs (45–60 kg BW) were used. Each pig was anesthetized and the head was orientated on the stereotaxic apparatus. A radiopaque dye was placed into the ventricle via a guide cannula inserted 3.5 mm forward of the bregma. A microcatheter was inserted into the third ventricle using ventriculography, and fixed with dental cement to a metal-mesh protector and screw anchors embedded into the skull. The opposite end of the microcatheter was externalized from the dorsal neck so that corticotropin-releasing hormone (CRH) could be injected easily. Simultaneously, a catheter was fitted in the jugular vein, and the free end of the catheter was externalized from the dorsal neck. Microcatheter-implanted pigs showed a normal progesterone concentration profile, and a constant cortisol level during at least two estrous cycles. Furthermore, intracerebroventricular injections of CRH (25 μg/500 μl) resulted in an increased plasma cortisol concentration ( P<0.05). Thus, the technique developed in this study enables us to approach the third ventricle in post-pubertal freely-moving pigs effectively over a long time, without causing stress.

Central administration of corticotropin-releasing hormone alters downstream movement in an artificial stream in juvenile chinook salmon ( Oncorhynchus tshawytscha)

We evaluated the effect of corticotropin-releasing hormone (CRH) on spatial distribution and downstream movement in an artificial stream in juvenile Chinook salmon ( Oncorhynchus tshawytscha) during the period when the fish were able to tolerate seawater. An intracerebroventricular (ICV) injection of CRH (500 ng) to hatchery fish significantly increased the proportion of fish that were distributed downstream of a mid-stream release site. A second group of hatchery fish were given ICV injections of saline (control) or CRH (500 ng) and released near the top of the stream. The time taken to enter a trap at the lower end of the stream was recorded. In all cases the groups given CRH had a higher proportion of fish that did not enter the trap within 60 min of release. However, in those fish that did enter the trap, treatment with CRH increased the speed of downstream movement to this point relative to control fish. Wild sub-yearling Chinook salmon were captured during their downstream migration to the estuary and given ICV injections of saline or CRH (500 ng) either 2, 3, or 7 days after transport from the river. As with hatchery fish, a significantly higher proportion of wild fish that were administered CRH did not enter the trap at the lower end of the stream. The mean time of passage for control fish decreased on each successive day (day 2 > day 3 > day 7). In contrast, the mean passage time of the wild fish that were given CRH was relatively constant through time, and was only significantly faster than control fish on day 2. The current study provides evidence that CRH alters the downstream movement of juvenile Chinook in a simulated stream environment, and produces behavioral effects similar to those of juvenile salmonids that are stressed during their downstream migration.