Lawrence M. Maness

Leptin enters the brain by a saturable system independent of insulin

Leptin, or OB protein, is produced by fat cells and may regulate body weight by acting on the brain. To reach the brain, circulating leptin must cross the blood-brain barrier (BBB). Intravenously injected radioiodinated leptin (125I-leptin) had an influx constant (Ki) into brain of (5.87)10(-4) ml/g-min, a rate 20 times greater than that of labeled albumin. Unlabeled leptin inhibited the influx of 125I-leptin in a dose-dependent manner whereas unlabeled tyrosine and insulin, which have saturable transport systems, were without effect. HPLC and acid precipitation showed that the radioactivity in brain and serum represented intact 125I-leptin. About 75% of the extravascular 125I-leptin in brain completely crossed the BBB to reach brain parenchyma. Autoradiography detected uptake at the choroid plexus, arcuate nuclei of the hypothalamus, and the median eminence. Saturable transport did not occur out of the brain. The results show that leptin is transported intact from blood to brain by a saturable system.

Decreased transport of leptin across the blood–brain barrier in rats lacking the short form of the leptin receptor☆ ☆

Leptin is produced in adipose tissue in the periphery, but its satiety effect is exerted in the CNS that it reaches by a saturable transport system across the blood-brain barrier (BBB). The short form of the leptin receptor has been hypothesized to be the transporter, with impaired transport of leptin being implicated in obesity. In Koletsky rats, the splice variant that gives rise to the short form of the leptin receptor contains a point mutation that results in marked obesity. We studied the transport of leptin across the BBB in Koletsky rats and found it to be significantly less than in their lean littermates. By contrast, Sprague-Dawley rats matched in weight to each of these two groups showed no difference in the blood-to-brain influx of leptin. HPLC showed that most of the leptin crossing the BBB in rats remained intact and capillary depletion showed that most of the leptin reached the parenchyma of the brain. The results indicate that the short form of the leptin receptor is involved in the transport of leptin across the BBB.

Peptides crossing the blood–brain barrier: some unusual observations

An interactive blood-brain barrier (BBB) helps regulate the passage of peptides from the periphery to the CNS and from the CNS to the periphery. Many peptides cross the BBB by simple diffusion, mainly explained by their lipophilicity and other physicochemical properties. Other peptides cross by saturable transport systems. The systems that transport peptides into or out of the CNS can be highly specific, transporting MIF-1 but not Tyr-MIF-1, PACAP38 but not PACAP27, IL-1 but not IL-2, and leptin but not the smaller ingestive peptides NPY, orexin A, orexin B, CART (55-102[Met(O)(67)]), MCH, or AgRP(83-132). Although the peptides EGF and TGF-alpha bind to the same receptor, only EGF enters by a rapid saturable transport system, suggesting that receptors and transporters can represent different proteins. Even the polypeptide NGF enters faster than its much smaller subunit beta-NGF. The saturable transport of some compounds can be upregulated, like TNF-alpha in EAE (an animal model of multiple sclerosis) and after spinal cord injury, emphasizing the regulatory role of the BBB. As has been shown for CRH, saturable transport from brain to blood can exert effects in the periphery. Thus, the BBB plays a dynamic role in the communication of peptides between the periphery and the CNS.

The neurotrophins and their receptors: Structure, function, and neuropathology

The neurotrophins are a family of polypeptides that promote differentiation and survival of select peripheral and central neurons. Nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3, neurotrophin-4, and neurotrophin-5 are included in this group. In recent years, tremendous advances have been made in the study of these factors. This has stimulated our review of the field, characterizing the neurotrophins from initial isolation to molecular analysis. The review also discusses their synthesis, localization, and responsive tissues, in both the periphery and CNS. The complex receptor interactions of the neurotrophins are also analyzed, as are putative signal transduction mechanisms. Discussion of the observed and postulated involvement in neuropathological disorders leads to the conclusion that the neurotrophins are involved in the function and dysfunction of the nervous system.

Permeability of the blood-brain barrier to amylin.

Amylin is co-secreted with insulin from the pancreas of patients with non-insulin dependent diabetes mellitus, and its deposition may contribute to the central nervous system (CNS) manifestations of this disease. Amylin, but not its mRNA, is found in brain, suggesting that CNS amylin is derived from the circulation. This would require amylin to cross the blood-brain barrier (BBB). We used multiple-time regression analysis to determine the unidirectional influx constant (Ki) of blood-borne, radioactively labeled amylin (I-Amy) into the brain of mice. The Ki was 8.99(10(-4)) ml/g-min and was not inhibited with doses up to 100 micrograms/kg, but it was inhibited by aluminum (Al). About 0.11 to 0.13 percent of the injected dose of I-Amy entered each gram of brain. Radioactivity recovered from brain and analyzed by HPLC showed that the majority of radioactivity taken up by the brain represented intact I-Amy. Capillary depletion confirmed that blood-borne I-Amy completely crossed the BBB to enter the parenchymal/interstitial fluid space of the cerebral cortex. Taken together, these results show that blood-borne amylin has access to brain tissue and may be involved in some of the CNS manifestations of diabetes mellitus.

Passage of human amyloid β-protein 1–40 across the murine blood-brain barrier

Abstract Previous studies have suggested that the amyloid β-protein present in the brains of patients with Alzheimers disease may be derived in part from peripheral blood. We determined that after IV injection of synthetic amyloid β-protein 1–40 (Aβ), labeled with radioactive 125 I(I-Aβ), radioactivity accumulated in the brains of mice by a nonsaturable mechanism. Radioactivity also accumulated in the brain after the IV injection of radioiodinated reverse amyloid β-protein 40-1 (I-rAβ). Capillary depletion techniques, however, showed I-Aβ to have a much greater degree of association with brain capillaries than I-rAβ. Acid precipitation of radioactivity in CSF samples and recovery from cortical homogenates suggested the presence of intact I-Aβ within the CNS after peripheral administration. HPLC analysis of cortical homogenates confirmed the presence of intact I-Aβ. Gel electrophoresis of the CSF acid precipitates and of the HPLC fractions further verified the presence of intact blood-derived I-Aβ peptide in CNS. These results suggest that endogenous bloodborne Aβ can enter the CNS after associating with the capillary endothelium to accumulate intact within the parenchymal and CSF spaces of the brain.

Persistence of blood-to-brain transport of leptin in obese leptin-deficient and leptin receptor-deficient mice.

In lean CD-1 mice, leptin is delivered into the brain by a saturable transport mechanism. Previous work has shown that obesity is associated with decreased leptin transport. Here, we investigated the transport of leptin across the blood-brain barrier (BBB) in two murine models of obesity. Radioiodinated leptin was intravenously injected into ob/ob (no leptin production) and db/db (high leptin levels, but no long-form leptin receptor) mutant mice and their lean controls. In all groups, the labeled polypeptide was transported across the BBB by a saturable mechanism. The rates of transport were not significantly different between the mutant strains and their lean controls. The results demonstrate that leptin transport persists in the absence of production of the endogenous polypeptide or its signal-transducing receptor and suggest that the impaired transport previously seen is not directly explained by only obesity or alterations in serum plasma levels.

Selective transport of blood-borne interleukin-1α into the posterior division of the septum of the mouse braIn

Film autoradiography was used to demonstrate the transport and sites of accumulation of blood-borne radioiodinated interleukin-1 alpha (II-1 alpha) and other cytokines into the brain after intravenous administration. [125 I]Il-1 alpha, [125I]Il-1 beta, [125I]interleukin-1 receptor antagonist (II-1ra), and [125I]tumor necrosis factor-alpha (TNF alpha) labeled the choroid plexus and the capillary network 30 min after injection into the blood, suggesting that these areas may serve as sites of blood-to-brain transport. [125I]Il-1alpha, but not [125I]Il-1beta, [125I]Il-1ra, [125I]TNF alpha, or [125I]interleukin-2 (Il-2), was also found localized to the caudal region of the septal nuclei. Only unlabeled II-1 alpha was able to inhibit this accumulation. These findings provide further evidence for the passage of select cytokines across the blood-brain barrier (BBB) and are the first to identify a target site within the central nervous system (CNS) for a transported cytokine.

Antibodies to beta-amyloid decrease the blood-to-brain transfer of beta-amyloid peptide.

Amyloid-β peptides (Aβ) play an important role in the pathophysiology of dementia of the Alzheimers type and in amyloid angiopathy. Aβ outside the CNS could contribute to plaque formation in the brain where its entry would involve interactions with the blood-brain barrier (BBB). Effective antibodies to Aβ have been developed in an effort to vaccinate against Alzheimers disease. These antibodies could interact with Aβ in the peripheral blood, block the passage of Aβ across the BBB, or prevent Aβ deposition within the CNS. To determine whether the blocking antibodies act at the BBB level, we examined the influx of radiolabeled Aβ (125I-Aβ1-40) into the brain after ex-vivo incubation with the antibodies. Antibody mAb3D6 (élan Company) reduced the blood-to-brain influx of Aβ after iv bolus injection. It also significantly decreased the accumulation of Aβ in brain parenchyma. To confirm the in-vivo study and examine the specificity of mAb3D6, in-situ brain perfusion in serum-free buffer was performed after incubation of 125I-Aβ1-40 with another antibody mAbmc1 (DAKO Company). The presence of mAbmc1 also caused significant reduction of the influx of Aβ into the brain after perfusion. Therefore, effective antibodies to Aβ can reduce the influx of Aβ1-40 into the brain.

Saturable entry of ciliary neurotrophic factor into brain

Ciliary neurotrophic factor (CNTF), like tumor necrosis factor-alpha (TNF) and granulocyte-macrophage colony-stimulating factor (GM-CSF), is a cytokine with neurotrophic properties. Since all three cytokines are found in the periphery as well as brain, and since TNF and GM-CSF cross the blood-brain barrier (BBB) by a saturable mechanism, we investigated whether CNTF also saturably enters the brain from the blood. We found that CNTF crosses the BBB rapidly, with a rate of entry (Ki) of 4.60 (+/-0.78) x 10(-4) ml/g min, considerably faster than that of the 99mTc-albumin control. The Ki was reduced more than 3-fold by addition of excess unlabeled CNTF. The results indicate that CNTF is saturably transported across the BBB from blood to brain.