basilen blue as a proposed additive to cryoprotectant solutio...

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Date: Sun, 29 Apr 2007 19:55:40 -0700 (PDT)
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Subject: basilen blue as a proposed additive to cryoprotectant solutio...

[Basilen blue exerts powerful neuroprotection against ischemia, and other
toxic insults.]

Neuroscience. 2003;120(1):85-98.
Up-regulation of P2X2, P2X4 receptor and ischemic cell death: prevention
by P2 antagonists.
    Cavaliere F, Florenzano F, Amadio S, Fusco FR, Viscomi MT, D'Ambrosi
N, Vacca F, Sancesario G, Bernardi G, Molinari M, Volonte C. IRCCS Santa
Lucia Foundation, Rome, Italy.
    In the present work we examined the involvement of selected P2X
receptors for extracellular ATP in the onset of neuronal cell death
caused by glucose/oxygen deprivation. The in vitro studies of organotypic
cultures from hippocampus evidenced that P2X2 and P2X4 were up-regulated
by glucose/oxygen deprivation. Moreover, we showed that ischemic
conditions induced specific neuronal loss not only in hippocampal, but
also in cortical and striatal organotypic cultures and the P2 receptor
antagonists basilen blue and suramin prevented these detrimental
effects. In the in vivo experiments we confirmed the induction of P2X
receptors in the hippocampus of gerbils subjected to bilateral common
carotid occlusion. In particular, P2X2 and P2X4 proteins became
significantly up-regulated, although to different extent and in different
cellular phenotypes. The induction was confined to the pyramidal cell
layer of the CA1 subfield and to the transition zone of the CA2 subfield
and it was coincident with the area of neuronal damage. P2X2 was
expressed in neuronal cell bodies and fibers in the CA1 pyramidal cell
layer and in the strata oriens and radiatum. Intense P2X4
immunofluorescence was localized to microglia cells. Our results indicate
a direct involvement of P2X receptors in the mechanisms sustaining cell
death evoked by metabolism impairment and suggest the use of selected P2
antagonists as effective neuroprotecting agents.
PMID: 12849743

Neuropharmacology. 1999 Sep;38(9):1335-42.
Neuroprotective effects of modulators of P2 receptors in primary culture
of CNS neurones.
    Volonte C, Ciotti MT, D'Ambrosi N, Lockhart B, Spedding M. Institute
of Neurobiology, C.N.R., I.R.C.C.S. Santa Lucia, Rome, Italy.
    In previous studies (Volonte and Merlo, 1996. J. Neurosci. Res. 45,
183-193) basilen blue was shown to be a P2 receptor antagonist which
abrogated glutamate-mediated cytotoxicity in cerebellar neurones in
primary culture. Our work has now been extended to evaluate the
neuroprotective action of the compound in additional neuronal systems, as
well as in a different paradigm of cell death. We show that basilen blue
prevents L-glutamate-mediated neurotoxicity in rat cerebellar (90-100%
inhibition), cortical (60-70%) and hippocampal (50%) neurones. Similarly,
glutamate-dependent progressive darkening of cell bodies, loss of
phase-brightness and rapid cellular swelling are inhibited. Basilen blue
is significantly less toxic and more effective at blocking L-glutamate
toxicity in mixed cortical/glial cultures, compared to its structural
analogue cibacron blue. Moreover, its neuroprotective effect is correlated
with the time of incubation with granule neurones. Other purinoceptor
ligands, including 2,2'-pyridylisatogen, but not
pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid 4-sodium, are also
effective in preventing glutamate toxicity. Furthermore, basilen blue
prevents serum deprivation- and low potassium-induced apoptotic cell
death in cerebellar granule neurones. In summary, our data extend and
reinforce the possibility of a potential therapeutic use of P2 receptor
modulators as neuroprotective agents for the central nervous system.
PMID: 10471087

Neurochem Int. 2001 Mar;38(3):189-97.
Glucose deprivation and chemical hypoxia: neuroprotection by P2 receptor
antagonists.
    Cavaliere F, D'Ambrosi N, Ciotti MT, Mancino G, Sancesario G,
Bernardi G, Volonte C. Fondazione Santa Lucia, Via Ardeatina 306, 00179,
Rome, Italy.
    In this work we investigate cell survival after glucose deprivation
and/or chemical hypoxia and we analyse the neuroprotective properties of
selected antagonists of P2 ATP receptors. We find that in rat cerebellar
granule neurones, the antagonist basilen blue prevents neuronal death
under hypoglycaemia. Basilen blue acts through a wide temporal range and
it retains its efficacy under chemically induced hypoxic conditions, in
the presence of the respiratory inhibitors of mitochondria electron
transport chain complexes II (3-nitropropionic acid) and III (antimycin
A). In spite of the presence of these compounds, basilen blue maintains
normal intracellular ATP levels. It furthermore prevents neuronal death
caused by agents blocking the mitochondrial calcium uptake (ruthenium
red) or discharging the mitochondrial membrane potential (carbonyl
cyanide m-chlorophenylhydrazone). Inhibition of poly
(ADP-ribose) polymerase, modulation of the enzyme GAPDH and mitochondrial
transport of mono-carboxylic acids are not conceivable targets for the
action of basilen blue. Survival is sustained by basilen blue also in CNS
primary cultures from hippocampus and in PNS sympathetic-like
neurones. Partial neuroprotection is furthermore provided by three
additional P2 receptor antagonists: suramin,
pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid 4-sodium and
4,4'-diisothiocyanatostilbene-2,2'disulphonic acid. Our data suggest the
exploitation of selected P2 receptor antagonists as potential
neuroprotective agents.
PMID: 11099776

J Neurosci Res. 1996 Jul 15;45(2):183-93.
Selected P2 purinoceptor modulators prevent glutamate-evoked cytotoxicity
in cultured cerebellar granule neurons.
    Volonte C, Merlo D. Institute of Neurobiology, CNR, Rome, Italy.
    Primary cultures of granule neurons derived from cerebella of
postnatal rats are endowed with Glu receptors. Glu receptor agonists
exert a trophic influence on differentiating granule cells but, with
maturation, the cells become vulnerable to excitatory amino acids. Here
we show that the P2 purinoceptor antagonist basilen blue abolishes in rat
cerebellar granule neurons the cytotoxic action of glutamate with an IC50
in the 10-20 microM range. Within the same concentrations, basilen blue
inhibits binding of [3H] ATP to cerebellar granule cells,
glutamate-evoked release (but not uptake) of [3H] D-aspartate and Ca2+
uptake. Furthermore, the extracellular phosphorylation of a major 45-kDa
endogenous ecto-protein substrate of cerebellar granule neurons is
inhibited with an IC50 of about 1 microM. Similar effects are elicited by
5-adenylylimidodiphosphate, a P2 purinoceptor agonist, when supplied to
the neurons for 8 days previously to the addition of glutamate. Our data
point to the use of P2 purinoceptor modulators as novel elements for
understanding and controlling glutamate-mediated excitatory neurotoxicity
and neurotransmission. We suggest a possible involvement of P2
purinoceptors in these actions.
PMID: 8843035

[Would basilen blue help with tinnitus?]

Hear Res. 1994 Aug;78(2):181-8.
ATP antagonists cibacron blue, basilen blue and suramin alter
sound-evoked responses of the cochlea and auditory nerve.
    Kujawa SG, Fallon M, Bobbin RP. Kresge Hearing Research Laboratory of
the South, Department of Otorhinolaryngology and Biocommunication,
Louisiana State University Medical Center, New Orleans 70112.
    The P2-purinergic receptor antagonists suramin, cibacron blue and
basilen blue, the latter two being isomers of reactive blue 2, were
studied for their effects on sound-evoked responses from the cochlea
(cochlear microphonic, CM; summating potential, SP; distortion product
otoacoustic emissions, DPOAE) and auditory nerve (compound action
potential, CAP). Local application of these compounds (10-1000
microM) into the cochlear perilymph was associated with
concentration-dependent response alterations. Effects of suramin on
cochlear responses were minimal: High-intensity SP was reduced
slightly at concentrations > or = 330 microM without significant
alterations in CM or DPOAEs. The amplitude of the auditory nerve CAP was
suppressed and its latency increased at drug concentrations > or = 100
microM. Cibacron blue and basilen blue were of greater potency in their
effects on cochlear and auditory nerve responses. DPOAEs were generally
reduced, low-intensity SP was reduced and high-intensity SP was increased
and CM was little affected at drug concentrations 100-1000 microM. The
CAP was suppressed and its latency increased at concentrations > or = 33
microM. Effects of suramin were largely reversible; those associated with
cibacron blue and basilen blue generally were not. To the extent that
these drugs acted selectively as antagonists of ATP receptor-mediated
activity, results support the hypothesis that endogenous ATP exerts
profound actions at the level of the cochlea and the auditory nerve.
PMID: 7982811

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