Very low prevalence of bovine immunodeficiency virus infection in western Canadian cattle.

Bovine immunodeficiency virus (BIV) is a lentivirus that causes disease in cattle. Despite the large cattle industry in western Canada, the presence of BIV has not been examined to date. Genomic DNA, derived from semen and buffy coat samples, was analyzed by nested polymerase chain reaction (PCR) using specific primers for the gag, pol, and env genes of BIV. Despite utilizing a procedure that detected a minimum of 10 proviral copies, BIV sequences were not amplified in any of 317 buffy coat and 50 semen samples that were obtained from an archive that included 27 cattle breeds, collected from different sources in Alberta (1980-1999). In the 367 DNA samples examined, there was no evidence of BIV infection, suggesting that the prevalence of BIV infection was very low.

Hetero-concatemeric KIR6.X4/SUR14 channels display distinct conductivities but uniform ATP inhibition.

K(IR)6.1 and K(IR)6.2 are the pore-forming subunits of K(NDP)(,) the nucleotide-diphosphate-activated K(ATP) channels, and classical K(ATP) channels, respectively. "Hybrid" channels, in which the structure is predetermined by concatemerizing K(IR)6.1 and K(IR)6.2, exhibit distinct conductivities specified by subunit number and position. Inclusion of one K(IR)6.2 is sufficient to open K(IR)6. X-X-X-X/SUR1(4) in the absence of nucleotide stimulation through sulfonylurea receptor-1 (SUR1). ATP inhibited the spontaneous bursting of hybrid channels with an IC(50(ATP)) approximately 10(-)(5) m, similar to that of K(IR)6.2(4)-containing channels. These findings and a transient increase in K(NDP) channel activity following rapid wash-out of MgATP suggested that K(IR)6.1 is not ATP-insensitive as previously believed. We propose that SUR-dependent, inhibitory ATP-enhanced interactions of the cytoplasmic domains of both K(IR)6.1 and K(IR)6.2 stabilize a closed form of the M2 bundle in the gating apparatus.

[Effect of thermal treatment on total dietetic fiber, soluble and insoluble contents in legumes]. / Efecto del tratamiento térmico sobre el contenido de fibra dietética total, soluble e insoluble en algunas leguminosas.

During the seventies it was proposed that the deficient intake of dietetic fiber contributed both to the overweight and the suffering of brain vascular and digestive diseases. At the moment, its beneficial effect (which is proportional to the type of fiber consumed) has been widely demonstrated for its action on related diseases. The purpose of this paper was to prove that thermal treatment modifies the content of Total Dietetic Fiber (TDF). Soluble Dietetic Fiber (SDF) and Insoluble Dietetic Fiber (IDF) in lentil (Lens culinaris) and black beans (Phaseolus vulgaris), both species of high intake in the country, especially in Caracas. The determination of TDF and its fractions (SDF and IDF) was performed through the application of the gravimetric enzymatic method (21). The results indicate significant differences (Kruskal-Wallis Test 95% of confidence) in the contents of TDF and its fractions, between the crude legumes and those subject to thermal treatment. The IDF was predominant in all cases and according to its positive effects-apparently favors the consumers of these legumes. On the basis of these findings, it is considered important to continue the analysis of the TDFs constituents generated after the cooking of these legumes.

Discrimination between apo and iron-loaded forms of transferrin by transferrin binding protein B and its N-terminal subfragment.

Many pathogens of the Pasteurellaceae and Neisseriaceae possess a surface receptor that binds transferrin (Tf) as an initial step in an iron acquisition process. This receptor is comprised of two proteins, transferrin binding protein A (TbpA) and transferrin binding protein B (TbpB). Since the ability to recognize the iron-loaded form of Tf preferentially would be a useful attribute of these receptors, we examined this property in a number of bacterial species. In solid-phase binding assays with isolated membranes, only the receptor from Moraxella catarrhalis was capable of preferentially binding iron-loaded Tf. In a competitive affinity isolation assay which enabled us to resolve TbpA and TbpB, TbpA from all tested species was shown to bind both apo and iron-loaded Tf. Under these assay conditions TbpB from M. catarrhalis, Haemophilus somnus and Pasteurella haemolytica discriminated between apo and holo Tf, whereas TbpB from Neisseria meningitidis showed no discrimination. The ability of TbpB from N. meningitidis to bind iron-saturated hTf preferentially became evident in a TbpA- background or by using recombinant TbpB. In binding assays with recombinant fusion proteins, both intact TbpB and the N-terminal half of TbpB from all the tested species preferentially bound Fe-loaded Tf, indicating that this may be a conserved mechanism by which these organisms optimize their ability to acquire iron.

Characterization of the Pasteurella haemolytica transferrin receptor genes and the recombinant receptor proteins.

The tbpA and tbpB genes encoding the transferrin receptor proteins, TbpA and TbpB, from Pasteurella haemolytica A1 were cloned, sequenced and expressed in Escherichia coli. The genes were organized in a putative operon arrangement of tbpB- tbpA. The tbpB gene was preceded by putative promoter and regulatory sequences, and followed by a 96 base pair intergenic sequence in which no promoter regions were found, suggesting that the two genes are coordinately transcribed. The deduced amino acid sequences of the TbpA and TbpB proteins had regions of homology with the corresponding Neisseria meningitidis, N. gonorrhoeae, Haemophilus influenzae and Actinobacillus pleuropneumoniae Tbp and Lbp proteins. The intact tbpB gene was expressed in a T7 expression system and the resulting recombinant TbpB protein retained the functional bovine transferrin binding characteristics. The availability of the recombinant TbpB enabled us to demonstrate its specificity for ruminant transferrins, its ability to bind both the C-and N-terminal lobes of bovine transferrin and its preference for the iron-loaded form of this protein. Several attempts at expressing the cloned tbpA gene were unsuccessful, suggesting that the product of the gene may be toxic to E. coli.

Hypothalamic and extrahypothalamic sauvagine-like immunoreactivity in the bullfrog (Rana catesbeiana) central nervous system.

In the present study, immunocytochemistry and radioimmunoassay were used to investigate the presence of sauvagine in both hypothalamic and extrahypothalamic areas of the central nervous system (CNS) of the bullfrog (Rana catesbeiana) using a specific antiserum raised against synthetic non-conjugated sauvagine (SVG), a frog (Phyllomedusa sauvagei) skin peptide of the corticotropin-releasing factor (CRF) family. Sauvagine-immunoreactive (SVG-ir) bipolar neurons were found in the nucleus of the fasciculus longitudinalis medialis located in the rostral mesencephalic tegmentum. In the tectal mesencephalon, beaded SVG-ir fibres were present in the optic tectum, and in the torus semicircularis. Abundant SVG-ir varicose fibres were seen in the granulosa layer of the cerebellum, the nucleus isthmi, and the obex of the spinal cord. SVG-ir fibres were also seen by the alar plate of the rombencephalon. In the diencephalon, the antiserum stained parvocellular neurons of the preoptic nucleus (PON) which extended their dendrites into the cerebro-spinal fluid (CSF) of the third ventricle and projected their ependymofugal fibres to the zona externa (ZE) of the median eminence. Immunopositive fibres were also present in the medial forebrain bundle at the chiasmatic field, the posterior thalamus, the pretectal gray, and the ventrocaudal hypothalamus. In the telencephalon (forebrain), SVG-ir fibres were seen in the medial septum, the lateral septum, and the amygdala. The SVG immunoreactivity could not be detected after using the SVG antiserum previously immunoabsorbed with synthetic SVG (0.1 microM), but immunoblock of the antiserum with sucker (Catostomus commersoni) urotensin I (sUI), sole (Hippoglossoides elassodon) urotensin I, sucker CRF, rat/human CRF, or ovine CRF (0.1-10 microM) did not eliminate visualization of the immunoreactivity. In radioimmunoassay, the SVG antiserum did not crossreact with sUI, or the SVG fragments SVG1-16, SVG16-27, and SVG26-34, but it recognized the C-terminal fragment SVG35-40. Crossreaction with mammalian ovine CRF and rat/human CRF was negligible. Both hypothalamic and mesencephalic extracts gave parallel displacement curves to SVG. The results suggest the presence in the bullfrog brain of a SVG-like neuropeptide, i.e., a peptide of the CRF family, that either is SVG or shares high homology with the C-terminus of that peptide. The function of this neuropeptide in amphibians is not known at this time, but based on its anatomical distribution to the ZE it could affect the release of adrenocorticotropin (ACTH) or other substances from the amphibian pars distalis. Involvement of the SVG-like peptide in behavioural (forebrain), visual (thalamus-tegmentum mesencephali-pretectal gray-optic tectum), motor coordination (cerebellum), and autonomic (spinal) functions, as well as an undefined interaction with the CSF in the bullfrog, seems likely.

[Lipoprotein(a) concentrations in type I diabetes mellitus]. / Concentraciones de la lipoproteína (a) en la diabetes mellitus tipo I.

OBJECTIVES: To determine the distribution of lipoprotein(a) levels and prevalence of hyperLp(a) in diabetes mellitus type I (IDDM). To analyze the effect of glycemic control and microalbuminuria on Lp(a) levels. METHODS: Cross-sectional study of 263 subjects with IDDM with a mean age of 19.2 +/- 11.6 years and an evolutive course of 6.3 +/- 6.5 years. Apart from Lp(a), measurements were obtained from serum levels of lipids, apolipoproteins AI and B, fructosamine, glycosilated hemoglobin (HbA1c), and albuminuria in all patients. RESULTS: Mean serum concentrations of Lp(a) were 16.5 +/- 18.1 mg/dl and 18.5% of patients had Lp(a) levels > 30 mg/dl [hyperLp(a)]. With a multivariate regression analysis, the only variable correlated with Lp(a) levels was cholesterol LDL (p < 0.001). Patients with hyperLp(a) did not differ from the other patients in any of the other variables analyzed, and patients with HbA1c higher and lower than 8% had similar mean serum Lp(a) concentrations (14.0 +/- 16 vs. 17.9 +/- 20). Lp(a) concentration was also similar among patients with albuminuria higher and lower than 20 micrograms/min (16.6 +/- 20 vs. 17.7 +/- 16). CONCLUSIONS: Control of glycemia and microalbuminuria have no effect on Lp(a) concentrations in diabetes mellitus type I. HyperLp(a) is not indicative of a poor glycemic control.

Presence of sauvagine-like epitopes in the interrenal gland of the bullfrog Rana catesbeiana.

Immunocytochemistry was used to investigate the presence of corticotropin-releasing factor-like peptides in the interrenal (adrenal) glands of the bullfrog Rana catesbeiana by using specific antisera raised against synthetic nonconjugated rat/human corticotropin-releasing factor, urotensin I, and sauvagine. From these three antisera, covering a broad range of corticotropin-releasing factor-like immunoreactivities, only the sauvagine antiserum gave positive immunoreactivity. Sauvagine immunoreactivity was found in cortical cells grouped into cords in the renal zone of the interrenal gland. The central and subcapsular cords were less stained. Tyrosine hydroxylase-positive chromaffin cells were not sauvagine-immunoreactive. The immunoreactivity was abolished, in all cases, by previous immunoabsorption of the sauvagine antiserum with synthetic sauvagine (0.1 microM), but it was not eliminated by sucker (Catostomus commersoni) urotensin I, sole (Hippoglossoides elassodon) urotensin I, sucker corticotropin-releasing factor, rat/human corticotropin-releasing factor, or ovine corticotropin-releasing factor (0.1-10 microM). In a sauvagine radioimmunoassay, interrenal extracts displaced 125I-sauvagine from antiserum only partially, and not in parallel with the sauvagine standard curve. The results suggest that the sauvagine immunoreactivity in the R. catesbeiana interrenal gland may represent a novel sauvagine-like peptide.

Sequence, genetic analysis, and expression of Actinobacillus pleuropneumoniae transferrin receptor genes.

The tbpA and tbpB genes encoding the transferrin receptor proteins Tbp1 and Tbp2 from a serotype 7 strain of Actinobacillus pleuropneumoniae were cloned, sequenced, and expressed in Escherichia coli. The tbpB gene was preceded by putative promoter and regulatory sequences and was separated from the downstream tbpA gene by a 13 bp intercistronic sequence suggesting that the two genes may be coordinately transcribed. Determination of the nucleotide sequence of this region facilitated PCR amplification of the tbp region from a serotype 1 strain for comparative purposes. The deduced amino acid sequences of the Tbp1 proteins had regions of homology with Neisseria Lbp and Tbp1s and with TonB-dependent outer membrane (OM) receptors of E. coli. The deduced amino acid sequences of the Tbp2 proteins were nearly identical to those presented in previous studies. Upon high-level expression of the tbpA gene, a large proportion of the recombinant Tbp1 was found in inclusion bodies and could not be affinity-isolated with immobilized porcine transferrin. Most of the remaining expressed Tbp1 was present in the OM fraction, was expressed at the surface of E. coli cells, and retained binding activity that was specific for the C-lobe of porcine transferrin. Although recombinant Tbp2 was found in inclusion bodies during high-level expression, a significant proportion was associated with a novel OM fraction that appeared in sucrose density gradients which was distinct from the OM fraction containing recombinant Tbp1. The recombinant Tbp2 was accessible at the surface yet was unable to bind porcine transferrin. In contrast to previous observations, the binding by recombinant Tbp2 was specific for the C-lobe of porcine transferrin. These results indicate that the A. pleuropneumoniae transferrin receptor proteins have similar properties to the receptor proteins in Neisseria spp. and Haemophilus influenzae, and that functional studies performed with recombinant receptor proteins need to consider differences in processing and export of these proteins when expressed in heterologous hosts.

Angiotensinogen-like epitopes are present in the CNS of Aplysia california and co-localize with urotensin I- and urotensin II-like immunoreactivities in the cerebral ganglia.

Immunohistochemistry was used to demonstrate urotensin I (UI), urotensin II (UII), and angiotensinogen (Ao)-like immunoreactivities (ir) in the CNS of Aplysia californica. The fish UI is a 41 amino acid peptide that has 50% identity with mammalian corticotropin-releasing factor (CRF). Identity also exists between UI and angiotensinogen in a tetrapeptide at the N-terminus. Ao-ir neurones were found in the F cluster of the Aplysia cerebral ganglia. Beaded Ao-ir fibres were seen in the neuropile and commissure of the cerebral, pleural and pedal ganglia. Ao neurosecretory material was also seen in the perineural region of the proximal supralabial nerve. Previously we have demonstrated UI and UII immunoreactivities were present in the CNS of Aplysia. A comparison of adjacent sections of the cerebral ganglia immunostained sequentially for UI, UII and Ao revealed that all three immunoreactivities co-existed in the same cells of the F cluster of the cerebral ganglia. Liquid-phase immunoabsorption of the Ao antiserum revealed that porcine or human angiotensinogen but not UI or UII were able to quench Ao immunostaining. Conversely UI and UII staining were quenched by white sucker (Catatomus commersoni) UI and goby (Gillichtys mirabilis) UII, respectively, but they were not modified by angiotensinogen. These results suggest that UI-, UII-, and Ao-like peptides might co-exist as separate entities in the cerebral ganglia of Aplysia californica where they can act in an integrated and/or independent modulatory way.

Comparative localization of corticotropin and corticotropin releasing factor-like peptides in the brain and hypophysis of a primitive vertebrate, the sturgeon Acipenser ruthenus L.

The sturgeon is a primitive actinopterigian fish that, unlike modern teleosts, possess a portal vascular system that connects a true median eminence with the anterior pituitary as in mammals. The occurrence and localization of corticotropin and corticotropin releasing factor-like immunoreactivies were examined in the brain of the sturgeon (Acipenser ruthenus L.) by immunocytochemistry with antisera raised against synthetic non-conjugated human corticotropin, and rat/human corticotropin releasing factor. In the hypothalamus, corticotropin-immunoreactive parvicellular perikarya were found in the infundibular nucleus and in dendritic projections to the infundibular recess. In addition, ependymofugal corticotropin-immunoreactive fibres were found to terminate in the ventral hypothalamus. Corticotropin releasing factor-immunoreactive neurons were found in the rostral portion of the ventral hypothalamus (tuberal nucleus), and in the vicinity of the rostral aspect of the lateral recess. These cells projected to the dorsal hypothalamus, the ventral hypothalamus, the median eminence, the anterior and posterior telencephalon, the tegmentum mesencephali, and the pars nervosa of the pituitary. An affinity-purified UI antiserum failed to stain the sturgeon hypothalamus. Corticotrophs in the rostral pars distalis of the pituitary were also corticotropin-immunoreactive. In the neurointermediate lobe, only about 50% of cells of the pars intermedia appeared to be corticotropin-positive, the rest appeared unstained. These results suggest that the presence of corticotropin-like and corticotropin releasing factor-like peptides in the brain is a relatively early event in vertebrate evolution, already occurring in Chondrostean/Actinopterigian fishes, as exemplified by A. ruthenus. The close spatial relationship between corticotropin releasing factor immunoreactivity and corticotropin immunoreactivity in the ventral hypothalamus of A. ruthenus supports a possible interaction between the two systems in that area of the sturgeon brain. The pars intermedia might be an important site for corticotropin synthesis, even though the possibility cannot be excluded that the antiserum was recognizing the proopiomelanocortin molecule. The occurrence of corticotropin releasing factor immunoreactivity in the region of median eminence/pars intermedia of the sturgeon suggests that the sturgeon corticotropin releasing factor might regulate the adenohypophyseal release of proopiomelanocortin products in the same manner as in other vertebrates. The presence of extrahypothalamic corticotropin releasing factor-immunoreactive projections suggests further neuromodulatory functions for this peptide in A. ruthenus.

Localization of vasopressin-like immunoreactivity in the CNS of Aplysia californica.

Chromatographic and immunological evidence indicates that a vasopressin-like peptide might be present in the CNS of Aplysia californica, and that this peptide may be involved in modulating the behaviour of the gill. Immunocytochemical techniques using antisera raised against various vasopressin-like peptides were used to localize the sites containing these peptides in the CNS of Aplysia. Vasopressin-like immunoreactivity was found to be restricted to one single neuron in the abdominal ganglion and two small neurons located bilaterally in each pedal ganglion. Immunoreactive fibres were present in the neuropile of the abdominal, pedal, pleural and cerebral ganglia, but not in the buccal ganglion. The identification of these neurons provides a morphological localization for vasopressin-like substances detected previously in CNS extracts of Aplysia californica. In addition, the possibility of electrophysiological studies involving the immunoreactive neurons identified in the present paper will allow a more direct approach to study the physiological role of vasopressin-like peptides in Aplysia.

Distribution and coexistence of urotensin I and urotensin II peptides in the cerebral ganglia of Aplysia californica.

Urotensin I (UI) and urotensin II (UII) were demonstrated in the cerebral ganglia of Aplysia californica by applying immunocytochemical and radioimmunoassay procedures. Sequential analysis of adjacent sections of the cerebral ganglia of Aplysia demonstrated that the UI-immunoreactive (UI-IR) neurons of the F cluster of the cerebral ganglia also contained UII immunoreactivity (UII-IR). Both UI-IR and UII-IR were also observed in a cuff-like arrangement of fibers surrounding the proximal portion of the supralabial nerve, as well as in a few fibers in the anterior tentacular nerves. The UI-IR perikarya of the cerebral ganglia appeared to project to the entire CNS of Aplysia, but the UII-IR fibers appeared only in the neuropile and commissure of the cerebral ganglia. The UI-IR staining was abolished by previous immunoabsorption of the UI antiserum with sucker (Catastomus commersoni) UI, but not with ovine corticotropin-releasing factor (CRF), rat/human CRF, or goby (Gillichthys mirabilis) UII. Immunostaining with UII antiserum was quenched by goby UII, but not by sucker UII-A, UII-B, UII-A(6-12), or carp (Cyprinus carpio) UII-alpha and UII-gamma. The UII staining was not abolished by UI or somatostatin. The F cluster was not stained when a somatostatin antiserum was applied. Radioimmunoassay of dilutions of cerebral ganglia extract, using UII antiserum, revealed a parallel displacement curve to synthetic goby UII.(ABSTRACT TRUNCATED AT 250 WORDS)

Urotensin I-like immunoreactivity in the central nervous system of Aplysia californica.

In the present study the occurrence and localization of urotensin I (UI, a corticotropin releasing factor-like peptide) in the CNS of Aplysia californica were investigated by immunocytochemistry and radioimmunoassay. The RIA cross-reactivity pattern indicated that the UI antiserum used recognized an epitope in the C-terminal region of the UI, but it did not cross-react with mammalian corticotropin-releasing factor (CRF) and partially recognized sauvagine (SVG, a frog CRF-like peptide). The use of CRF-specific and sauvagine-specific antisera failed to give positive immunostaining. The application of UI antiserum (which does not cross-react with CRF in RIA) gave a positive staining, which was blocked by synthetic sucker (Catostomus commersoni) UI, but not by rat/human CRF (10 microM). On the basis of immunostaining and RIA parallel to fish UI displacement curves of cerebral ganglia extracts, the unknown UI/CRF-like substance in the Aplysia ganglia is likely to have greater homology with sucker UI than with the known CRF peptides. Urotensin I-immunoreactive (UI-ir) neurons were seen mainly in the F neuron clusters, located in the midline and rostrodorsal portion of the cerebral ganglia. Few UI-ir neurons were also found in the C and D neuron clusters of the cerebral ganglia, as well as in the left pleural and abdominal ganglia. In addition, numerous fine and coarse, and beaded UI-ir fibers were found in the cerebral commissure. UI-ir fibers were also seen in the neuropile of the buccal, pedal and pleural ganglia, and abdominal ganglion. A cuff-like arrangement of UI-ir fibers was seen in the supralabial nerves.(ABSTRACT TRUNCATED AT 250 WORDS)

Identification and characterization of a porcine-specific transferrin receptor in Actinobacillus pleuropneumoniae.

All six strains of Actinobacillus pleuropneumoniae screened for the ability to use different transferrins as a source of iron for growth were capable of using porcine but not human, bovine, or avian transferrins. A specific binding activity for porcine transferrin (pTf) was expressed in cells grown in the presence of specific iron-chelators and was repressed by addition of excess iron. Two iron-repressible outer-membrane proteins of 105 and 56 kD were specifically isolated from serotype 1, 2 and 7 strains of A. pleuropneumoniae by an affinity-isolation method using biotinylated porcine transferrin and streptavidin-agarose.

Receptors for transferrin in pathogenic bacteria are specific for the host's protein.

Transferrin receptors detected by a solid-phase binding assay were shown to be specific for the host's transferrin in the representative bacterial pathogens Neisseria meningitidis (human), Pasteurella haemolytica (bovine), and Actinobacillus pleuropneumoniae (porcine). Consistent with the receptor specificity, iron-deficient bacteria were only capable of utilizing transferrin from the host as a source of iron for growth.

Comparison of the abilities of different protein sources of iron to enhance Neisseria meningitidis infection in mice.

This study was done primarily to determine whether the previously observed specificity of the meningococcal transferrin and lactoferrin receptors for human proteins was maintained in vivo during meningococcal infection in mice. Preliminary experiments evaluating the choice of host strain, the age and sex of mice, and the growth conditions of the meningococci indicated that 45-day-old female Swiss Webster mice challenged with meningococci grown on low-pH, low-iron Mueller-Hinton agar plates were appropriate for this study. The comparison of transferrins and lactoferrins from different species demonstrated that only the human forms of these proteins were utilized by meningococci; there was significantly greater mortality among mice treated with iron-saturated human transferrin or lactoferrin (93 and 100%, respectively) than among those not treated or treated with iron-saturated bovine transferrin or bovine lactoferrin (0%). Provision of exogenous hemoglobin also resulted in increased mortality, although not as great as that observed with amounts of transferrin with equivalent iron content, which parallels the more effective utilization of transferrin and lactoferrin in in vitro growth experiments. In addition, unlike with transferrin and lactoferrin, there was no difference in utilization of human and bovine hemoglobin in vitro or in vivo.

Sauvagine-like and corticotropin-releasing factor-like immunoreactivity in the brain of the bullfrog (Rana catesbeiana).

Immunocytochemical methods were used to investigate the occurrence and distribution of sauvagine, corticotropin-releasing factor-, or urotensin I-like immunoreactivities (SVG-ir, CRF-ir, UI-ir, respectively) in the bullfrog (Rana catesbeiana) brain, using specific antisera raised against non-conjugated SVG, ovine CRF, rat/human CRF, and UI. In the hypothalamus, SVG-ir was found in the magnocellular perikarya, in the dorsal and ventral regions of the preoptic nucleus, and in the hypothalamo-hypophyseal projections to the external zone as well as the internal zone of the median eminence, to pars nervosa, and in fibres running from the pars nervosa to the pars intermedia of the pituitary. In contrast, CRF-ir was found only in parvocellular perikarya, mainly localized in the rostro-ventral part of the preoptic nucleus, with fine processes protruding through the ependyma of the third ventricle, fibre projections terminating in the anterior preoptic area and in the neuropil of the periventricular gray, and a caudal projection to the external zone of the median eminence. No CRF-ir staining was seen in the pars nervosa and pars intermedia. The use of UI-specific antisera failed to give a positive response in the frog brain. It is concluded that, in the frog brain, two anatomically different CRF-like (or SVG-like) systems co-exist, comparable to the reported co-existence of UI-ir and CRF-ir neuronal systems in fish brain.