![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
TOXICOLOGY
Section on Developmental and Molecular Pharmacology, Laboratory of Developmental Neurobiology (D.E.B., J.M.H., D.A.) and Pregnancy and Perinatology Branch (C.Y.S.), National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland; and Department of Clinical Biochemistry (A.P., T.G., I.G.), Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel
Received for publication
December 9, 2003
Accepted
March 8, 2004.
 |
Abstract |
|---|
 Top Abstract Materials and MethodsResultsDiscussionReferences |
|---|
-amyloid peptide (25-35). In the present study, all D-amino acid peptides of NAPVSIPQ and SALLRSIPA were synthesized and compared respectively to the corresponding all L-amino acid peptides. In rat cerebral cortical test cultures cotreated with 1 µM tetrodotoxin, the D-amino acid peptides produced similar potency and efficacy for neuroprotection as that observed for their respective L-amino acid peptides. Since all these peptides tested individually exhibited attenuation of efficacy at concentrations of >10 pM, combinations of these peptides were tested for possible synergies. Equimolar D-NAPVSIPQ and D-SALLRSIPA combination treatment produced potent neuroprotection (EC50, 0.03 fM) that did not attenuate with increasing concentrations. Similarly, the combination of L-NAPVSIPQ and D-SALLRSIPA also had high potency (EC50, 0.07 fM) without attenuation of efficacy. Combined administration of peptides was tested in a model of fetal alcohol syndrome and in a model of learning impairment: apolipoprotein E knockout mice. Intraperitoneal administration of D-NAPVSIPQ plus D-SALLRSIPA to pregnant mice (embryonic day 8) attenuated fetal demise after treatment with an acute high dose of alcohol. Furthermore, oral administration of D-NAPVSIPQ plus D-SALLRSIPA significantly increased fetal survival after maternal alcohol treatment. Apolipoprotein E knockout mice injected with D-NAPVSIPQ plus D-SALLRSIPA showed improved performance in the Morris water maze. These studies suggest therapeutic potential for the combined administration of neuroprotective peptides that can act through a mechanism independent of chiral recognition.
) and activity-dependent neurotrophic factor (ADNF) (Brenneman and Gozes, 1996). Both of these glial proteins are neuroprotective and regulated by vasoactive intestinal peptide, a polypeptide shown to have growth-promoting and neurotrophic properties in vitro and in vivo (Brenneman and Eiden, 1986; Gressens et al., 1993). NAPVSIPQ and SALLRSIPA are synthesized peptides that exhibited neuroprotection at femtomolar concentrations (Gozes and Brenneman, 2000). Previous structure-activity studies with either an alanine scan (Wilkemeyer et al., 2003) or conservative substitutions (Brenneman et al., 1998) have shown a complete loss of neuroprotective activity with the replacement of single amino acids. In cerebral cortical cell cultures, these peptides have been shown to prevent neuronal cell death associated with tetrodotoxin (electrical blockade), the -amyloid peptide (the Alzheimer's disease neurotoxin), N-methyl-D-aspartate (excitotoxicity), and gp120, the human immunodeficiency virus envelope protein (Brenneman et al., 1998; Bassan et al., 1999). In addition, NAPVSIPQ and SALLRSIPA have been shown to provide protection from oxidative stress in pheochromocytoma and neuroblastoma cell lines (Offen et al., 2000; Steingart et al., 2000). Recent studies have shown that these peptides can be protective as antagonists of ethanol inhibition of L1-mediated cell-cell adhesion (Wilkemeyer et al., 2002). Furthermore, these peptides have been shown to be protective in vivo. For example, daily injections of SALLRSIPA or NAPVSIPQ to newborn apolipoprotein E-deficient mice accelerated the acquisition of developmental reflexes and, in the case of NAPVSIPQ, prevented short-term memory deficits (Bassan et al., 1999). Intranasal administration of NAPVSIPQ or SALLRSIPA promoted learning and memory (Gozes et al., 2000, 2002), whereas subcutaneous injection of NAPVSIPQ protected against edema formation, behavioral deficits, and death in a model of closed head injury (Beni-Adani et al., 2001). Intravenous injection of NAPVSIPQ after mid-cerebral artery occlusion (a model for stroke in rodents) resulted in protection against apoptosis, smaller infarct size, and behavioral improvements (Leker et al., 2002). Furthermore, pretreatment with an equimolar mixture of SALLRSIPA and NAPVSIPQ resulted in protection against fetal demise and growth retardation associated with fetal alcohol syndrome (FAS) (Spong et al., 2001). In the present study, the neuroprotective action of NAPVSIPQ and SALLRSIPA in either the D- or L-enantiomer conformation was investigated in cerebral cortical cultures and various models of neurodegeneration and growth retardation. The results suggest that D-enantiomers of NAPVSIPQ and SALLRSIPA maintain femtomolar-acting neuroprotective activity in vitro and further show that combined treatment with these peptides in vivo has significant protective actions against severe oxidative stress and neurodegeneration. Because NAPVSIPQ and SALLRSIPA maintain their biological efficacy in the D-form, studies were performed that demonstrate for the first time that these peptides can elicit protection after oral administration.
The current studies have first used neuroprotection against tetrodotoxin as an in vitro screen to compare the D- and L-forms of NAPVSIPQ and SALLRSIPA. Electrical blockade was chosen as the model system to make these comparisons because of its relevance to the activity-dependent mechanism of development as well as its first use in detecting and characterizing the action of both activity-dependent neurotrophic factor and activity-dependent neuroprotective protein, the parent proteins for SALLRSIPA and NAPSVIPQ, respectively. The choices of in vivo models were based on precedent with previous studies that used a mouse model of fetal alcohol syndrome (Spong et al., 2001) and the apolipoprotein E knockout mice (Bassan et al., 1999) that had been shown to be responsive to the L-forms of at least one of these peptides. Therefore, the choice of experimental systems was based primarily on preceding studies; however, both the in vitro and in vivo systems share a common link with induced apoptosis. Combinations of the peptides are emphasized in the present study because the previous work with the fetal alcohol model indicated that both L-peptides were required for maximum protection from both fetal demise and growth restriction.
|
Materials and Methods |
|---|
|
TopAbstractMaterials and MethodsResultsDiscussionReferences |
|---|
Cell Culture. Rat cerebral cortical cell cultures were prepared by previously described methods (Hill et al., 1993). Briefly, dissociated cerebral cortical tissue was seeded on a confluent layer of astrocytes previously prepared from newborn rat brain cerebral cortex. Twentyfour hours later, fluorodeoxyuridine (15 µg/ml) was added to the cultures. Cultures were maintained in 5% horse serum in minimal essential medium supplemented with defined media components as previously described (Romijn et al., 1982). After 4 days of growth in vitro, the cultures were given a complete change of medium and treated once with peptide dissolved in phosphate-buffered saline. The duration of the treatment period was 4 days. To assess neuroprotection, cultures were cotreated with 1 µM tetrodotoxin (TTX), a model of apoptotic death that is relevant to neurons dependent on electrical activity for their survival (Brenneman et al., 1983). To terminate neuroprotection, the cells were fixed in 2.5% paraformaldehyde for immunocytochemistry, and neurons were identified with antibodies against neuron-specific enolase by previously described methods (Schmechel et al., 1978). Neuronal cell counts were performed on 30 fields (200x magnification) without knowledge of treatment.
Animal Models
FAS in Mice. A well established model for FAS was used to test the efficacy of the peptides in mice (Webster et al., 1980). This paradigm is a test for efficacy against severe oxidative stress produced from alcohol administration (Spong et al., 2001). The model was chosen because it allowed for a rapid and relevant evaluation of agents efficacious against severe oxidative stress and because of its relevancy to FAS. To assess the protective effects of the peptides, the number of fetal demises was determined. The animals were sacrificed at ED18 (10 days after peptide/alcohol treatment). The number of living fetuses, fetal demises, and total litter size (living fetuses plus demises) was determined. This allowed for the documentation that all litters began with the same number. In the surviving ED18 fetuses, fetal and fetal brain weights were evaluated to assess the protective effects of the peptides. C57Bl/6J female mice were kept under a 12-h light/dark regimen with food and water available at all times. The mice received humane animal care in compliance with the "Guidelines for the Care and Use of Experimental Animals." Sixweek-old females (21-24 g) were mated with C57Bl/6J males for 4 h. The presence of a vaginal plug was considered day 0 of pregnancy. Mice were intraperitoneally injected with 25% ethyl alcohol in saline (v/v) or vehicle alone at 0.03 ml/kg maternal body weight at 9:00 AM on ED8. Pretreatment with the study peptides was given 30 min prior to alcohol as described. D-NAPVSIPQ or L-NAPVSIPQ (0.5 mg) was dissolved in 50 µl dimethyl sulfoxide and diluted with filtered Dulbecco's phosphate-buffered saline (DPBS) to a final volume of 5 ml (concentration, 100 µg/ml); of this solution, 0.2 ml (200 µl) was given per animal. D-SALLRSIPA (1.0 mg) was dissolved in 1 ml of DPBS and diluted in filtered DPBS to a final volume of 10 ml (concentration, 100 µg/ml); of this solution, 0.2 ml (200 µl) was administered for the 20-µg dose. This solution was further diluted 1:10 in DPBS for the 2-µg dose. Dosages of the peptides were D-NAPVSIPQ (20 µg), D-SALLRSIPA (20 or 2 µg), and NAPVSIPQ (20 µg) plus D-SALLRSIPA (20 µg). Since the animals receiving alcohol were incapacitated for approximately 6 h following injection, food and water were withheld from all groups for the initial 6 h after injection to allow for an accurate assessment of fetal weights. The mean pup weight for each litter was calculated, with the litter mean used for all statistical analysis. Percent demises were calculated by dividing the number of demises by the total number of fetuses (live plus demises). Statistical analysis included analysis of variance for continuous variables, Mann-Whitney U for nonparametric data, chi square for categorical variables, or Fisher's exact test where appropriate, with p < 0.05 considered significant.
Apolipoprotein E Knockout Mice: Behavior Assay. Inheritance of apolipoprotein E4 is a major risk factor in Alzheimer's disease (Corder et al., 1993). These studies, along with the investigations of apolipoprotein E-deficient animals, indicated that an apolipoprotein E functioning system is required for repair functions (Ignatius et al., 1986). Memory deficits and cholinergic impairments have been described in adult apolipoprotein E-deficient mice. Furthermore, developing apolipoprotein E-deficient mice have been shown to be cognitively impaired (Gordon et al., 1995; Gozes et al., 1997). Peptides (0.5 mg each) were dissolved in 0.01 M acetic acid (30 µl) and further diluted with saline. Each of the test drugs (D-NAPVSIPQ or D-SALLRSIPA) (0.5 µl) was subcutaneously injected daily in 20 µl per mouse. This protocol was used for the first 4 days of life. From day 5 to 10, the amount of the peptides and the solution volume was doubled. From day 11 to 14, the amount of peptide injected was 2 µg each in 80 µl of saline per mouse. A week after the cessation of peptide injection, the animals were subjected to the Morris water maze as previously described (Bassan et al., 1999). Briefly, mice were subjected to two daily tests in a water maze, including a hidden platform. The experiment was conducted over a 4-day period. For the first test, both the platform and animal were situated in a new location with regards to the pool every day (with the pool being immobile). The experiment was performed as follows. The animal was positioned on the platform for 0.5 min and then placed in the water and allowed to find the platform for 90 s. After finding the platform or being placed on the platform for an additional 0.5 min, the animal was placed back in the water (in the previous position) for a second daily test; then the animal would search again for the hidden platform (retained in the previous position). The time required to reach the platform in the second trial was recorded, and it was indicative of short-term (working) memory. Significance was assessed by one way analysis of variance with Student Neuman-Keuls multiple comparison of means test.
|
Results |
|---|
|
TopAbstractMaterials and MethodsResultsDiscussionReferences |
|---|
10 pM (p < 0.05). Likewise, the D- and L-forms of NAPVSIPQ were very similar, each exhibiting a complex dose response with two apparent maxima (Fig. 2). The EC50 values were 0.01 fM and 2 pM for L-NAPVSIPQ and 0.07 fM and 2 pM for D-NAPVSIPQ. Efficacies at the higher potency peak were not significantly different between the peptides, and the maximum efficacy of the D-peptides differed by only 10%.
|
|
Combinations of peptides were also tested. For these experiments, the two peptides were given in equimolar amounts of each peptide at the amount indicated in each figure. In Fig. 3, the effect of D-NAPVSIPQ plus D-SALLRSIPA was shown to produce a different dose response from that observed with either agent alone. Significantly, there was no attenuation of the survival-promoting activity at higher concentrations of the combined D-peptides. The EC50 was very potent at 0.03 fM, and the maximal efficacy was evident for seven orders of magnitude. The number of surviving neurons after D-NAPVSIPQ plus D-SALLRSIPA treatment (0.1 fM) was not different from that of the control cultures. Thus, a broader therapeutic range of effective concentrations was apparent when the D-peptides were used in combination. Similar experiments conducted with both L-SALLRSIPA and L-NAPVSIPQ resulted in a >60% loss of maximum efficacy, with no change in potency compared with that observed after treatment with D-NAPVSIPQ plus D-SALLRSIPA.
|
Another series of experiments was conducted to show the effect of combining L- and D-forms of NAPVSIPQ and SALLRSIPA. As shown in Fig. 4, the treatment with L-NAPVSIPQ and D-SALLRSIPA resulted in full efficacy and high potency (EC50, 0.07 fM) in preventing apoptotic death of neurons treated with TTX. There was no apparent attenuation of the protective activity at high concentrations of peptide (>1 pM); i.e., synergy was again evident. In contrast, treatment with D-NAPVSIPQ and L-SALLRSIPA resulted in full efficacy but attenuation of the survival-promoting activity at concentrations of 0.1 pM. These data indicate that D-SALLRSIPA is the only peptide among the L- and D-peptides studied that produced no attenuation of survival-promoting activity when used in combination with any of the other peptides.
|
In Vivo Efficacy. The studies of the protective properties of D- and L-peptides of NAPVSIPQ and SALLRSIPA were extended to a mouse model of FAS and to apolipoprotein E-deficient mice, a model of cognitive impairment and neurodegeneration. In the FAS model, the efficacy of the peptides was assessed by counting the number of surviving fetuses. The effects of peptide treatments administered intraperitoneally are presented in Fig. 5. Treatment with alcohol resulted in a 37% fetal demise compared with 6% in controls. A 30-min pretreatment with either D-NAPVSIPQ or D-SALLRSIPA significantly reduced fetal demise compared with those treated with alcohol alone (p < 0.03). A combination of D-NAPVSIPQ and D-SALLRSIPA was also protective; however, a clear dose-dependence was observed, with 20 µg of each being effective and 2 µg of each not significantly increasing fetal survival compared with ethanol-treated controls. L-NAPVSIPQ and D-SALLRSIPA, a combination shown to be protective in the in vitro testing, also produced protection in the FAS model.
|
To extend the studies in the FAS model, higher doses were used to test whether the D-peptides were effective after oral administration (Fig. 6). After oral gavage, 40 µg of D-NAPVSIPQ was not effective; however, treatment with a combination of D-NAPVSIPQ and D-SALLRSIPA significantly increased the number of surviving fetuses compared with the group treated with ethanol alone. The body weights were also measured in the same experiment as shown in Fig. 7. Significant decreases in body weight were observed after the alcohol treatment compared with the control group; however, oral treatment with the D-peptides did not significantly increase the body weight compared with the fetuses from the mothers injected with alcohol alone. Treatment with D-NAPVSIPQ and D-SALLRSIPA alone had no effect on body weight compared with controls.
|
|
Performance of apolipoprotein E-deficient mice was tested in the Morris water maze to assess the effects of the D-peptides on short-term spatial memory. Increases in performance as measured by decreases in latencies to find a hidden platform were observed a week after the cessation of the two-week daily D-SALLRSIPA plus D-NAPVSIPQ injections (i.e., in 21-day-old mice exposed to a 4-day training protocol) (Fig. 8). Short-term memory processes were examined by performance in the water maze and measured by the time required to find the hidden platform in the second of two daily trials. The platform location and starting point at which the animal was placed in the water were held constant within each pair of daily trials, but both locations were changed every day. All groups of tested mice learned to find the hidden platform after 4 testing days, as assessed by comparing the latencies to find the hidden platform on the first and last trial days (p < 0.01); however, differences were already observed on the first day of testing, with apolipoprotein E-deficient mice treated with the peptide mixture finding the platform faster than saline-treated apolipoprotein E-deficient mice (p < 0.05). Further significant differences were observed on the 2nd day of testing between these two groups (p < 0.03).
|
|
Discussion |
|---|
|
TopAbstractMaterials and MethodsResultsDiscussionReferences |
|---|
). In the mid-cerebral artery occlusion paradigm, L-NAPVSIPQ was found to provide protection, whereas D-NAPVSIPQ (tested at a single dose) was not effective. These studies indicate that the protective actions of D-NAPVSIPQ are dose-dependent and may not completely predict L-NAPVSIPQ (Leker et al., 2002).
Although unusual, other examples of biologically active peptides have been reported to be equally efficacious and potent in both the all D- and all L-conformation. However, to our knowledge, the present study is the first report of neuroprotective action that does not exhibit a stereoselectivity. Several of the previous studies of nonchiral effects of peptides suggested membrane phenomena, for example, through the formation of pores, as in the case of the antibiotic magainin and cecropin A (Wade et al., 1990). The D-enantiomeric peptides of both of these peptides exhibit similar antibiotic and single channel conductance as their respective L-enantiomers. Another example of a peptide that produced biological actions equally well in the D- or L-form is the amyloidogenic peptide (A1-42) derived from the amyloid precursor protein that is believed to have a causative role in the neurodegeneration observed in Alzheimer's disease (Cribbs et al., 1997). Although it is not clear whether the toxic A peptide forms pores, there is a report of A1-40 forming a cation channel in model membranes (Arispe et al., 1993). In the case of A1-42, the toxicity associated with this peptide may be mediated by the cross-linking of receptors through fibrillary forms of this peptide. The formation of protofibrils and fibrils may not be dependent on chirality. A recent report indicates that L-NAPVSIPQ does bind to and inhibit A aggregation, suggesting structural interaction between these two peptides (Ashur-Fabian et al., 2003).
D-peptides have been shown to interact with chaperone systems. For example, D-peptides compete with L-peptides for the same binding site in DnaJ, and this interaction can inhibit refolding functions of this chaperone system (Bischofberger et al., 2003). In addition, all D- and all L-conformers of the functional element of 
A-crystallin do not show marked differences in their chaperone-like activity (Bhattacharyya and Sharma, 2001). To speculate, a chaperone-like function may also be affected by these peptides. The latter is of particular interest because SALLRSIPA is very similar to CALLRCIPA, a sequence found in the chaperone heat shock protein 60 (Gozes and Brenneman, 1996).
Recent studies indicate that D-peptides derived from chemokines can interact with the CXCR4 receptor (Zhou et al., 2002). Remarkably, the D-peptides act as antagonists to this receptor, resulting in inhibition of CXCR4-dependent human immunodeficiency virus-1. In the case of the chemokine-related peptides, the D-peptides exhibited either equivalent or greater potency than L-peptide counterparts; however, these actions were confined to an antagonist action. It is not clear whether NAPVSIPQ and SALLRSIPA are acting to stimulate a neuroprotective process or providing an inhibition of a function associated with neuronal cell death. If the model of the chemokine receptor were relevant to the present study, our results could be interpreted as antagonizing a proapoptotic pathway. There is no previous observation of a nonstereoselective agonist action that is receptor-mediated.
The fundamental finding of the present study is that two neuroprotective and structurally related peptides do not exhibit stereoselective constraints to produce their biological activity at extraordinarily high potencies. Whether a similar recognition site or common mode of entry account for the initial events of cellular interaction remains to be established. Previous studies have not provided a systematic comparison of actions between the two peptides, yet a diverse set of actions have been observed in central nervous system related preparations. Neuroprotective actions of SALLRSIPA have been associated with the regulation of calcium homeostasis (Glazner et al., 1998) and transcription factors (Glazner et al., 1998; White et al., 2000). A slightly longer form of SALLRSIPA (ADNF14) has been shown to be associated with stimulation of the mitogen-activated protein kinase and protein kinase C pathways (Gressens et al., 1999). NAPVSIPQ neuroprotection has been associated with stimulation of cGMP and nitric oxide (Ashur-Fabian et al., 2001), with the effects on cGMP being more pharmacologically relevant to the neuroprotective actions. Thus, the L-peptides of NAPVSIPQ and SALLRSIPA have multiple pharmacological effects on neuroprotection, with a unifying mechanism not yet apparent. Contrasting concentration-effect curves of the two peptides support distinctive mechanisms. Although both peptides exhibit extraordinary potencies at femtomolar concentrations on neurons in culture, NAPVSIPQ clearly has two peaks of neuroprotective activity, whereas SALLRSIPA has one. Interestingly, both peptides show less potent neuroprotective effects in enriched neuronal cultures (sparse number of glia), suggesting a glial involvement in the full potency neuroprotective effect (Brenneman et al., 1998; Zemlyak et al., 2000).
All of the peptides investigated in the current study have exhibited attenuating biological responses with either a single- or double-inverted U-shaped dose response. Although characteristic of many peptide responses, the mechanism(s) of these effects remains unclear. Alternative explanations of these attenuating responses with increasing concentration of peptide include receptor desensitization, ligand-ligand interactions, or activation of different and opposing mechanisms that are dose-dependent.
Exploration of the effects of each of the peptides alone and in combination has concentration-effect profiles that suggest synergistic rather than additive actions. Whereas it is clear that the L-forms of both NAPVSIPQ and SALLRSIPA behave similarly to their respective D-form in vitro, the combinations resulted in synergies with respect to the inverted U-shaped dose response (i.e., the attenuation of neuroprotective activity with increasing concentrations). It is apparent that all combinations involving D-SALLRSIPA resulted in a clear plateauing effect, with no loss of protective response with increasing concentrations. The broadening of the effective concentrations of these agents increases the probability of their potential clinical success by obviating the confounding property of concentration-dependent attenuation of protection. Regardless of the mechanism, we believe that the combined administration of these peptides has distinct pharmacological advantages, including neuroprotection elicited at high potency that is insensitive to the use of the D-isomeric form of NAPVSIPQ and SALLRSIPA. However, evidence of similar synergies from combined peptide treatment in vivo remains to be demonstrated.
The discovery of the activity of the D-peptides along with the demonstration that these peptides are orally active opens up a new arena for peptiditic therapeutic agents. Although larger amounts of peptide are required for biological activity through the oral route of administration compared with i.p. or intranasal routes, the advantages for drug applications are significant. As shown in Fig. 6, only oral treatment with D-SALLRSIPA (40 µg) alone prevented fetal demise, whereas the same amount of D-NAPVSIPQ did not significantly reduce the amount of fetal death compared with the alcohol group. Importantly, when given intraperitoneally, both D-peptides produced effective protection from fetal death compared with the alcohol group. These data indicate that the route of administration is an important variable in evaluating the action of the D-peptides and further suggest that both the individual and combination of peptides should be tested, as they differ from the protective activity observed with the L-peptides given intraperitoneally. As shown previously, both L-peptides also prevented growth restriction, whereas the individual L-peptides had no effect on fetal weight (Spong et al., 2001). In the present study, the D-peptides at the same dose did not prevent the loss of fetal weight (Fig. 7). Furthermore, previous studies with the same model indicated that at the same dosage (20 µg) as employed for the individual D-peptides in the present study, L-NAPVSIPQ was effective in preventing fetal death but L-SALLRSIPA was not (Spong et al., 2001). Treatment with each of the D-peptides produced a significant increase in fetal survival compared with the alcohol-treated group. The pharmacodynamic/pharmacokinetic responses that contribute to the protective actions of the D-peptides will be important next steps in characterizing their action. These initial studies did not reveal any gross toxic responses to the D-peptides, but systematic studies of toxicity, metabolism, and biodistribution from the D-peptides will also be necessary next steps for the advancement of these agents. The current studies are an important first step in our evaluation of neuroprotective D-peptides as therapeutic agents.
|
Footnotes |
|---|
ABBREVIATIONS: ADNP, activity-dependent neuroprotective protein; ADNF, activity-dependent neurotrophic factor; ED, embryonic day; FAS, fetal alcohol syndrome; TTX, tetrodotoxin; DPBS, Dulbecco's phosphate-buffered saline.
Address correspondence to: Douglas E. Brenneman, Drug Discovery, Johnson and Johnson Pharmaceutical Research and Development, L.L.C., Welsh and McKean Roads, Spring House, PA 19477. E-mail address: dbrennem{at}prdus.jnj.com
|
References |
|---|
|
TopAbstractMaterials and MethodsResultsDiscussionReferences |
|---|
Arispe N, Pollard HB, and Rojas E (1993) Giant multilevel cation channels formed by Alzheimer disease amyloid -protein [AP-(1-40)] in bilayer membranes. Proc Natl Acad Sci USA 90: 10573-10577.
Ashur-Fabian O, Giladi E, Furman S, Steingart RA, Wollman Y, Fridkin M, Brenneman DE, and Gozes I (2001) Vasoactive intestinal peptide and related molecules induce nitrite accumulation in the extracellular milieu of rat cerebral cortical cultures. Neurosci Lett 307: 167-170.[CrossRef][Medline]
Ashur-Fabian O, Segal-Ruder Y, Skutelsky E, Brenneman DE, Steingart R, Giladi E, and Gozes I (2003) The neuroprotective peptide NAPVSIPQ inhibits the aggregation of the beta-amyloid. Peptides 24: 1413-1423.[CrossRef][Medline]
Bassan M, Zamostiano R, Davidson A, Pinhasov A, Giladi E, Perl O, Bassan H, Blat C, Gibney G, Glazner G, et al. (1999) Complete sequence of a novel protein containing a femtomolar-activity-dependent neuroprotective peptide. J Neurochem 72: 1283-1293.[CrossRef][Medline]
Beni-Adani L, Gozes I, Cohen Y, Assaf Y, Steingart RA, Brenneman DE, Eizenberg O, Trembolver V, and Shohami E (2001) A peptide derived from activity-dependent neuroprotective protein (ADNP) ameliorates injury response in closed head injury in mice. J Pharmacol Exp Ther 296: 57-63.
Bhattacharyya J and Sharma KK (2001) Conformational specificity of mini-alphaA-crystallin as a molecular chaperone. J Pept Res 57: 428-434.[CrossRef][Medline]
Bischofberger P, Han W, Feifel B, Schonfeld HJ, and Christen P (2003) D-peptides as inhibitors of the DnaK/DnaJ/GrpE chaperone System. J Biol Chem 278: 19044-19047.
Brenneman DE and Eiden LE (1986) Vasoactive intestinal peptide and electrical activity influence neuronal survival. Proc Natl Acad Sci USA 83: 1159-1162.
Brenneman DE and Gozes I (1996) A femtomolar-acting neuroprotective peptide. J Clin Investig 97: 2299-2307.[Medline]
Brenneman DE, Hauser J, Neale E, Rubinraut S, Fridkin M, Davidson A, and Gozes I (1998) Activity-dependent neurotrophic factor: structure-activity relationships of femtomolar-acting peptides. J Pharmacol Exp Ther 285: 619-627.
Brenneman DE, Neale EA, Habig WH, Bowers LM, and Nelson PG (1983) Developmental and neurochemical specificity of neuronal deficits produced by electrical impulse blockade in dissociated spinal cord cultures. Dev Brain Res 9: 13-27.
Corder EH, Saunders AM, Strittmatter WJ, Schmechel DE, Gaskell PC, Small GW, Roses AD, Haines JL, and Pericak-Vance MA (1993) Gene dose of apolipoprotein-E type-4 allele and the risk of Alzheimer's disease in late onset families. Science (Wash DC) 261: 921-923.
Cribbs DH, Pike CJ, Weinstein SL, Velazquez P, and Cotman CW (1997) All-D-enantiomers of beta-amyloid exhibit similar biological properties to all-L-betaamyloid. J Biol Chem 272: 7431-7436.
Glazner GW, Boland A, Brenneman DE, Gozes I, and Mattson MP (1998) ADNF-9 protects neurons against oxidative stress-induced death by activation of the transcription factor NF-kappa B. Mol Biol Cell 9: 2049.
Glazner GW and Mattson MP (2000) Differential effects of BDNF, ADNF9 and TNF on levels of NMDA receptor subunits, calcium homeostasis and neuronal vulnerability to excitotoxicity. Exp Neurol 161: 442-452.[CrossRef][Medline]
Gordon I, Grauer E, Genis I, Sehayek E, and Michaelson DM (1995) Memory deficits and cholinergic impairments in apolipoprotein E-deficient mice. Neurosci Lett 199: 1-4.[CrossRef][Medline]
Gozes I, Alcalay R, Giladi E, Pinhasov A, Furman S, and Brenneman DE (2002) NAPVSIPQ accelerates the performance of normal rats in the water maze. J Mol Neurosci 19: 167-170.[Medline]
Gozes I, Bachar M, Bardea A, Davidson A, Rubinraut S, Fridkin M, and Giladi E (1997) Protection against developmental retardation in apolipoprotein E-deficient mice by a fatty neuropeptide: implications for early treatment of Alzheimer's disease. J Neurobiol 33: 329-342.[CrossRef][Medline]
Gozes I and Brenneman DE (1996) Activity-dependent neurotrophic factor (ADNF). An extracellular neuroprotective chaperonin. J Mol Neurosci 7: 235-244.[Medline]
Gozes I and Brenneman DE (2000) A new concept in the pharmacology of neuroprotection. J Mol Neurosci 14: 61-68.[CrossRef][Medline]
Gozes I, Giladi E, Pinhasov A, Bardea A, and Brenneman DE (2000) Activity-dependent neurotrophic factor: intranasal administration of femtomolar-acting peptides improves performance in a water maze. J Pharmacol Exp Ther 293: 1091-1098.
Gressens P, Hill JM, Gozes I, Fridkin M, and Brenneman DE (1993) Growth factor function of vasoactive intestinal peptide in whole cultured mouse embryos. Nature (Lond) 362: 155-158.[CrossRef][Medline]
Gressens P, Marret S, Bodenant C, Schwendimann L, and Evrard P (1999) Activity-dependent neurotrophic factor-14 requires protein kinase C and mitogen-associated protein kinase kinase activation to protect the developing mouse brain against excitotoxicity. J Mol Neurosci 13: 199-210.[CrossRef][Medline]
Hill JM, Mervis RF, Avidor R, Moody TW, and Brenneman DE (1993) HIV envelope protein-induced neuronal damage and retardation of behavioral-development in rat neonates. Brain Res 603: 222-233.[CrossRef][Medline]
Ignatius MJ, Gebicke-Harter PJ, Skene JH, Schilling JW, Weisgraber KH, Mahley RW, and Shooter EM (1986) Expression on apolipoprotein E during nerve degeneration and regeneration. Proc Nat Acad Sci USA 83: 1125-1129.
Leker RR, Teichner A, Grigoriadis N, Ovadia H, Brenneman DE, Fridkin M, Giladi E, Romano J, and Gozes I (2002) NAPVSIPQ, a femtomolar-acting peptide, protects the brain against ischemic injury by reducing apoptotic death. Stroke 33: 1085-1092.
Offen D, Sherki Y, Melamed E, Fridkin M, Brenneman DE, and Gozes I (2000) Vasoactive intestinal peptide (VIP) prevents neurotoxicity in neuronal cultures: relevance to neuroprotection in Parkinson's disease. Brain Res 854: 257-262.[CrossRef][Medline]
Romijn HJ, Habets AM, Mud MT, and Wolters PS (1982) Nerve outgrowth, synaptogenesis and bioelectric activity in rat cerebral cortex tissue cultured in serum-free chemically defined medium. Dev Brain Res 2: 140-146.[CrossRef]
Schmechel D, Marangos PJ, and Brightman J (1978) Neuron specific enolase is a molecular marker for peripheral and central neuroendocrine cells. Nature (Lond) 276: 834-836.[CrossRef][Medline]
Spong CY, Abebe DT, Gozes I, Brenneman DE, and Hill JM (2001) Prevention of fetal demise and growth restriction in a mouse model of fetal alcohol syndrome. J Pharmacol Exp Ther 297: 774-779.
Steingart RA, Solomon B, Brenneman DE, Fridkin M, and Gozes I (2000) VIP and peptides related to activity-dependent neurotrophic factor protect PC12 cells against oxidative stress. J Mol Neurosci 15: 137-145.[CrossRef][Medline]
Wade D, Boman A, Wahlin B, Drain CM, Andreu D, Boman HG, and Merrifield RB (1990) All D-amino acid-containing channel-forming antibiotic peptides. Proc Natl Acad Sci USA 87: 4761-4765.
Webster WS, Walsh DA, Lipson AH, and McEwen SE (1980) Teratogenesis after acute alcohol exposure in inbred and outbred mice. Neurobehav Toxicol 2: 227-234.
White DM, Walker S, Brenneman DE, and Gozes I (2000) CREB contributes to the increased neurite outgrowth of sensory neurons induced by vasoactive intestinal polypeptide and activity-dependent neurotrophic factor. Brain Res 868: 31-38.[CrossRef][Medline]
Wilkemeyer MF, Chen SY, Menkari CE, Brenneman DE, Sulik KK, and Charness ME (2003) Differential effects of ethanol antagonism and neuroprotection in peptide fragment NAPVSIPQ prevention of ethanol-induced developmental toxicity. Proc Natl Acad Sci USA 100: 8543-8548.
Wilkemeyer MF, Menkari CE, Spong CY, and Charness ME (2002) Peptide antagonists of ethanol inhibition of L1-mediated cell-cell adhesion. J Pharmacol Exp Ther 303: 110-116.
Zemlyak I, Furman S, Brenneman DE, and Gozes I (2000) A novel peptide prevents death in enriched neuronal cultures. Regul Pept 96: 39-43.[CrossRef][Medline]
Zhou NM, Luo ZW, Luo JS, Fan XJ, Cayabyab M, Hiraoka M, Liu DX, Han XB, Pesavento J, Dong CZ, et al. (2002) Exploring the stereochemistry of CXCR4-peptide recognition and inhibiting HIV-1 entry with D-peptides derived from chemokines. J Biol Chem 277: 17476-17485.
This article has been cited by other articles:
|
|
 |
|
  L. Visochek, R. A. Steingart, I. Vulih-Shultzman, R. Klein, E. Priel, I. Gozes, and M. Cohen-Armon PolyADP-Ribosylation Is Involved in Neurotrophic Activity J. Neurosci., August 10, 2005; 25(32): 7420 - 7428. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Divinski, L. Mittelman, and I. Gozes A Femtomolar Acting Octapeptide Interacts with Tubulin and Protects Astrocytes against Zinc Intoxication J. Biol. Chem., July 2, 2004; 279(27): 28531 - 28538. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
 |
 |
|
 |
 |
 |
