Levels of Corticotropin Releasing Factor-Like Immunoreactivity in Mammalian Hypothalamic and Extrahypothalamic Brain Tissue as Determined With a Monoclonal Antibody to the Ovine Material.

Hollander, Charles S.; Audhya, Tapan; Russo, Maria; Passarelli, Joseph; Nakane, Takeichiro; Schlesinger, David H.

doi:10.1210/endo-112-6-2206

Cited by 16 publications

(6 citation statements)

References 16 publications

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“…It was prepared by the same methods used previously to raise antiserum for a highly sensitive radioimmunoassay for CRF (22,23). As confirmed by logit-log transformations of the data and statistical comparisons of the resultant regression lines, this antiserum to oCRF displayed parallelism to extracts of guinea pig hypothalamus and extra-hypothalamic brain tissue (unpublished data).…”

Section: Methodsmentioning

confidence: 99%

Evidence for a role of endogenous corticotropin-releasing factor in cold, ether, immobilization, and traumatic stress.

Nakane¹,

Audhya²,

Kanie³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

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The role of corticotropin-releasing factor (CRF) in four model stresses (cold, ether, immobilization, and trauma) was examined in the guinea pig by using passive immunoneutralization with anti-CRF antiserum. Plasma corticotropin levels were measured at various times after exposure to stress, and groups treated with CRF antiserum were compared with those treated with normal rabbit serum. Of the four stresses tested, ether had the most pronounced effect on corticotropin secretion. Treatment with anti-CRF inhibited most of the ether-induced corticotropin secretory response, the difference between the normal serum-and the anti-CRF antiserum-treated groups being significant at 5 and 10 min (P < 0.01). Corticotropin responses to cold stress in the two groups differed at the 0.05 level of significance at 10 and 20 min. After administration of trauma (leg fracture), a statistically significant difference (P < 0.01) between the two groups also was evident, albeit only at 20 min. During immobilization, corticotropin levels differed significantly from control only in the normal serum-treated group but not in the anti-CRF-treated group. These findings show that CRF antiserum was effective in reducing corticotropin levels, indicating that CRF has an important role in mediating corticotropin response to stress. The fact that neutralization was incomplete might be due to an inability of the antiserum to sufficiently neutralize the endogenous CRF or, more likely, reflects the contribution of additional mediators, notably catecholamines and vasopressin, of corticotropin release upon stress.Corticotropin-releasing factor (CRF) has long been postulated to have a physiological role in modulating corticotropin (adrenocorticotropic hormone, ACTH) and l3-endorphin secretion (1-3). Recent elegant studies in the rat (4) strongly imply that CRF and/or an endogenous peptide immunologically related to CRF has a physiologic role in the maintenance of elevated levels of circulating corticotropin in adrenalectomized rats and in the activation of the pituitary-adrenal axis due to ether stress. However, diverse stresses differ considerably in their pathophysiology, sequelae, and underlying neural control of corticotropin secretion (5-20). There-fore, these studies were undertaken utilizing a similar in vivo paradigm in the guinea pig to ascertain whether CRF performs an analogous role in other forms of stress. The results of the passive immunoneutralization approach (4, 21), which in this instance entails intravenous administration of a potent antibody against ovine CRF (oCRF) to the guinea pig before exposure to cold (6,15,19), trauma (8, 9), immobilization (14, 15, 17), or ether stress (5, 10, 11, 18), suggest that an endogenous CRF-like peptide does play a physiological role in the regulation of corticotropin in these model stresses. METHODSA polyclonal antibody (hereafter referred to as anti-CRF) from rabbit 471 was used for the in vivo neutralization studies. It was prepared by the same methods used previously to raise antiserum fo...

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Section: Methodsmentioning

confidence: 99%

Evidence for a role of endogenous corticotropin-releasing factor in cold, ether, immobilization, and traumatic stress.

Nakane¹,

Audhya²,

Kanie³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

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“…This indicates the possibility of a particularly antigenic region at the C terminus, in agreement with the suggestion from structural prediction studies for ovine CRF-41 (Pallai, Mabilia, Goodman et al 1983) that hydrophilic residues 33, 34, 36 and 37 are likely to be involved in antigenic sites. Another murine mono¬ clonal antibody raised against ovine CRF-41 (Hollander, Audhya, Russo et al 1983) was reported to be able to bind ovine CRF-41 and sauvagine and was used to detect immunoreactivity in ovine, bovine and murine tissue, suggesting that this antibody also recognized a region of CRF-41 with cross-species homology. Biological characterization of the antibodies KCHMB001 and KCHMB003 showed that they neutralized the ACTH-releasing activity of rat CRF-41 in a pituitary bioassay in a manner similar to the monoclonal described by Van Oers et al (1989).…”

Section: Discussionmentioning

confidence: 99%

Production and utilization of monoclonal antibodies to human/rat corticotrophin-releasing factor-41

Milton

Hillhouse²,

Nicholson³

et al. 1990

Journal of Molecular Endocrinology

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Murine monoclonal antibodies against human/rat corticotrophin-releasing factor-41 (CRF-41) were produced and characterized for use in the immunological and biological characterization of CRF-41. Spleen cells from BALB/c mice immunized with CRF-41 conjugated to bovine gamma-globulin were fused with a BALB/c-derived non-secretor X-63 myeloma line. Hybridomas were selected for CRF antibody production by enzyme-linked immunosorbent assay, and positive hybridomas cloned twice. Three monoclonal antibodies were obtained (KCHMB001, KCHMB002 and KCHMB003) and characterized as IgG1, IgG1 and IgG2a isotypes respectively, with affinity constants for rat CRF-41 of 30, 53 and 34 nmol/l respectively. All three monoclonal antibodies recognize an epitope contained between residues 34 and 41 of the human/rat sequence. The antibodies were able to neutralize the ACTH-releasing activity of rat CRF-41, applied to rat pituitary fragments in vitro, in a dose-dependent manner. Isoelectric focusing showed that KCHMB003 detected bands of synthetic rat CRF-41 and rat [Met(O)21,38]-CRF-41 at pH 7.1 and 6.8 respectively. Use of KCHMB003 in a two-site enzyme-amplified immunoassay showed that this antibody recognizes both synthetic rat CRF-41 and immunoreactive CRF-41 in rat hypothalamic tissue extracts.

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“…Sections of the L6/S1 spinal cord, brainstem, and hypothalamus were immunostained with two different antibodies that were generated in rabbits against CRF (Hollander et al, 1983; Silverman et al, 1989; 1:1,500 and 1:1,000 dilution, respectively; gifts from Dr. Ann‐Judith Silverman, Columbia University, and Dr. Wylie Vale, The Salk Institute). Sections were incubated in the primary antibody for 48 hr, washed in 0.01 M phosphate‐buffered saline (PBS), and incubated for 80 min in goat anti‐rabbit IgG conjugated to Alexa 594 or Alexa 488 (Molecular Probes, Eugene, OR).…”

Section: Methodsmentioning

confidence: 99%

Distribution and origin of corticotropin‐releasing factor‐immunoreactive axons in the female rat lumbosacral spinal cord

Puder

Papka

2001

J of Neuroscience Research

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Corticotropin-releasing factor (CRF) is a neuropeptide traditionally known for its hormonal role in the hypothalamic/pituitary/adrenal stress axis. However, CRF has been reported in axons in sites that may be considered outside of the direct stress axis, e.g., in axons in the lumbosacral spinal cord associated with the micturition response. Whether any of these CRF-immunoreactive axons interacts with uterine-related preganglionic autonomic neurons or projection neurons in the lumbosacral spinal cord is unknown. Thus, immunohistochemistry and retrograde tracing were employed to determine the presence, distribution, and origin of CRF-immunoreactive axons in the L6/S1 spinal cord of the female rat and to ascertain whether these axons are associated with uterine-related neurons. CRF-immunoreactive axons were present in the dorsal horn, medial and lateral collateral pathways, dorsal intermediate gray, laminae VlI and X, and sacral parasympathetic nucleus of the spinal cord. Nitric oxide-synthesizing, i.e., NADPH-d-positive neurons and pseudorabies virus labeled uterine-related neurons were in the sacral parasympathetic nucleus and were closely apposed by CRF-immunoreactive axons. Injection of retrograde tracers (fluorogold or fast blue) into the L6/S1 spinal cord labeled neurons in the hypothalamic paraventricular nucleus and pontine Barrington's nucleus, and some of these neurons were immunoreactive for CRF. This study demonstrates that CRF-immunoreactive axons are present in the L6/S1 spinal cord of the female rat in areas associated with sensory and autonomic processing. Some of these axons originate from the paraventricular nucleus and Barrington's nucleus and are adjacent to uterine-related neurons. These results indicate that CRF may influence neural activity related to the female reproductive system.

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Levels of Corticotropin Releasing Factor-Like Immunoreactivity in Mammalian Hypothalamic and Extrahypothalamic Brain Tissue as Determined With a Monoclonal Antibody to the Ovine Material.

Cited by 16 publications

References 16 publications

Evidence for a role of endogenous corticotropin-releasing factor in cold, ether, immobilization, and traumatic stress.

Evidence for a role of endogenous corticotropin-releasing factor in cold, ether, immobilization, and traumatic stress.

Production and utilization of monoclonal antibodies to human/rat corticotrophin-releasing factor-41

Distribution and origin of corticotropin‐releasing factor‐immunoreactive axons in the female rat lumbosacral spinal cord

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