Immunoreactive dynorphin in pituitary and brain.

Goldstein, Avram; Ghazarossian, Vartan E.

doi:10.1073/pnas.77.10.6207

Cited by 156 publications

(57 citation statements)

References 23 publications

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“…The distribution of ir-Dyn across tissues in the toad is generally similar to that reported for the rat (4)(5)(6)(7). For (4,5,7,14).…”

Section: Discussionsupporting

confidence: 79%

“…For (4,5,7,14). In toad neural tissue, the relative contribution of each molecular size ir-Dyn to the total immunoreactivity varies from tissue to tissue in a pattern different from that seen in the rat.…”

Section: Discussionmentioning

confidence: 99%

“…Using antiserum ("Lucia") raised against dynorphin-(1-13) (D13) and specific for the sequence dynorphin- (3)(4)(5)(6)(7)(8)(9)(10)(11)(12) (3), immunoreactive dynorphin (ir-Dyn) has been found in pig, beef, and rat pituitary and rat brain and spinal cord (4)(5)(6)(7). This paper describes the presence of ir-Dyn in toad (Bufo marinus) pituitary, brain, and spinal cord.…”

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confidence: 99%

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A dynorphin-like opioid in the central nervous system of an amphibian.

Cone¹,

Goldstein²

1982

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

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We have provided evidence for the existence of a biologically active opioid in toad (Bufo marinus) brain that is immunoreactive with antiserum raised against dynorphin-(1-13). Compared with porcine dynorphin, this opioid is similar in apparent molecular weight on the basis of gel permeation chromatography and is more hydrophobic on the basis of high-performance liquid chromatography. After purification, its opioid biological activity was demonstrated on the guinea pig ileum myenteric plexus-longitudinal muscle preparation. It was found to be less potent, and to have a similar sensitivity to antagonism by naloxone, in comparison with porcine dynorphin. Because it is immunoreactive with antiserum specific for porcine dynorphin, it probably has considerable sequence homology. Generally, the tissue distribution ofimmunoreactive dynorphin in the toad is similar to that in the rat, with highest concentrations in the neurointermediate lobe of the pituitary. However, the anterior lobe of the toad pituitary contains considerably lower concentrations than are found in the rat anterior lobe. There appear to be three size classes of immunoreactive dynorphin in toad neural tissue, each with apparent molecular weight below 12,000, similar to the size classes of immunoreactive dynorphin found in pig and rat. However, in toad spinal cord (and possibly in brain) there is immunoreactive dynorphin of greater apparent molecular weight, which has not been reported in mammalian tissue. The contribution of each molecular size to the total immunoreactivity varies from tissue to tissue and is different from that observed in the rat.

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“…The distribution of ir-Dyn across tissues in the toad is generally similar to that reported for the rat (4)(5)(6)(7). For (4,5,7,14).…”

Section: Discussionsupporting

confidence: 79%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

A dynorphin-like opioid in the central nervous system of an amphibian.

Cone¹,

Goldstein²

1982

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

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“…On the otherhand, leucine-enkephalin RIA (not shown) does detect material that cochromatographs with authentic leucine-enkephalin. Thus, it is possible to conclude that both peptides exist in posterior pit tuitary, a result consonant with that of Goldstein and Ghazarossian (23). The relatively equal concentrations of leucine-enkephalin and dynorphin immunoreactivities suggest that our dynorphin demonstration is not due to crossreactivity with leucine-enkephalin, because a large excess of leucine-enkephalin _ A t was required to block the demonstration.…”

Section: Methodssupporting

confidence: 82%

Dynorphin immunocytochemical localization in brain and peripheral nervous system: preliminary studies.

Watson¹,

Akil²,

Ghazarossian³

et al. 1981

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

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151

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Dynorphin is a recently discovered peptide belonging to the family of endogenous opioids (1). The first five amino acids at the NH2 terminus of dynorphin are identical to leucine-enkephalin; however, from position 6 to 13 it has a unique structure (the remainder ofthe peptide is currently unknown). While the anatomical localization of 3-endorphin and the enkephalins have been well described (2-19), little is currently known about the distribution of dynorphin. This peptide was extracted from porcine pituitaries and was purified by means ofa bioassay using the guinea pig ileum. We therefore elected to begin our studies on dynorphin immunocytochemistry by looking at rat pituitary and brain and at guinea pig ileum. We also extended our studies to guinea pig adrenal because of the previous reports of an enkephalin-like immunoreactivity in that organ. MATERIALS AND METHODSTissue preparation for immunocytochemistry was as described (17). Antisera against dynorphin-(1-13) were prepared, with the resulting antigenic specificity largely directed toward a COOHterminal region (20). Several antisera were produced, with similar specificities; that of highest titer (Lucia) was chosen, because it has been the most thoroughly characterized for radioimmunoassay (RIA). As published elsewhere (20), the Lucia antiserum is used in RIA at titers of from 1:50,000 to 1:200,000and crossreacts with leucine-enkephalin less than 10-8, and [3H]leucine-enkephalin does not appear to bind even in dilutions as low as 1:30. For peroxidase-antiperoxidase immunocytochemistry this antiserum was used at a 1:600 dilution for a 24-hr incubation at 40C (peroxidase-antiperoxidase and related reagents were from Sternberger-Meyer Immunocytochemical, Jarrettsville, MD)..An important issue in the immunocytochemical study of dynorphin distribution in brain is the potential confusion with leucine-enkephalin-containing structures. The antiserum used in this study (Lucia) was raised against dynorphin-(1-13) (Peninsula Labs, San Carlos, CA) and its antigenic determinant includes residues 4 and 5 of dynorphin and leucine-enkephalin. While there is little crossreactivity with leucine-enkephalin under RIA conditions, the Lucia antiserum was used in a much more concentrated form for immunocytochemistry. It is therefore possible that the lower-affinity populations ofantibody may come into play under our immunocytochemical conditions, and that the crossreactivity with other peptides, including leucineenkephalin, may be different from that observed in RIA. For that reason complete control blocking studies were carried out on all tissues, using 2-20 A.M concentrations ofmethionine-and leucine-enkephalin, 8-endorphin, oxytocin, vasopressin, and dynorphin-(1-13).In experiments in which relative concentrations ofdynorphin and enkephalin were determined, a combined high-performance liquid chromatography (HPLC)-RIA procedure was employed. All tissue was obtained fresh after decapitation, kept chilled on ice during dissection, then frozen immediately on dry ice and ...

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“…lmmunoreactive dynorphin has been shown to be distributed in the central nervous system, the pituitary gland and peripheral nervous system (3)(4)(5). A synthetic dynorphin-(1-13) (Dyn) is potent as natural dynorphin- (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17) in bioassay (6), and it has been shown to produce analgesia and catalepsy (7), alteration in centrally stimulated gastric acid secretion (8) and spontaneous feeding (9). However, the physiological role of dynorphin is at present uncertain.…”

mentioning

confidence: 99%