Health Effects of Penitrem A – Neurotoxic Issues

 

 

This page lists medical journal articles discussing the association between neurotoxic issues and the mycotoxin known as penitrem A.

The Health Effects of Penitrem A page of the Paradigm Change site provides further information on the effects of this mycotoxin.

 

Berntsen H. F., Wigestrand M. B., Bogen I. L., Fonnum F., Walaas S. I., Moldes-Anaya A.. Mechanisms of penitrem-induced cerebellar granule neuron death in vitro: possible involvement of GABAA receptors and oxidative processes. Neurotoxicology. 2013;35:129–136. PMID: 23313729 

The fungal neurotoxin penitrem A penetrates the blood-brain-barrier and causes cerebellar pathology in rats, including mild effects on granule neurons. The aim of the current study was to investigate the potential toxicity of penitrem A in rat cerebellar granule neurons in vitro. Cerebellar granule cells were exposed to penitrem A, alone or together with different pharmacological agents, before cell survival was assessed with the MTT assay or formation of reactive oxygen species (ROS) was investigated with the DCF assay. Penitrem A caused a time- and concentration-dependent reduction in cell survival, as well as a concentration-dependent increase in ROS production. Co-incubation with diazepam, GABA, BAPTA-AM, vitamin E, SP600125 and cyclosporine A significantly reduced cell death. Our results show that penitrem A is toxic to cerebellar granule neurons in vitro. Further, ROS production and the GABAA receptor are likely to be involved in the induction of neuronal death following penitrem A exposure. A disruption of calcium homeostasis and activation of the JNK pathway may also play a role in penitrem A neurotoxicity.

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Moldes-Anaya Angel, Rundberget Thomas, Fæste Christiane K., Eriksen Gunnar S., Bernhoft Aksel. Neurotoxicity of Penicillium crustosum secondary metabolites: tremorgenic activity of orally administered penitrem A and thomitrem A and E in mice. Toxicon. 2012;60:1428–1435. PMID: 23085423

In the present study, the lowest tremor-inducing dose after single oral administration of penitrem A to mice was 0.50 mg/kg bw. The estimated half maximal effective dose (ED(50)) in respect to the visual tremor scale was 2.74 mg/kg bw. Mice receiving the maximum penitrem A dose (8 mg/kg bw) suffered severe spontaneous tremors and even convulsions. Thomitrem A and E are penitrem analogues lacking the C-16-C-18 ether linkage and possessing an olefin at C-18-C-19. Compared with penitrem A, the lowest tremor-inducing dose of thomitrem A was 16-times higher (8 mg/kg bw) and thomitrem E was found to be non-tremorgenic at the highest dose tested (16 mg/kg bw). During a recovery phase of two weeks post administration animals appeared restored and no changes in feeding and other biological processes were observed. An initial dose-related weight reduction was observed 2 days after penitrem A administration. Penitrem A was absorbed and distributed to gastrointestinal tract, liver, kidneys and brain in the mice. Elimination of penitrem A appeared to be mainly hepatic and the highest concentration levels were found 1 h post administration for all investigated organs. The relationship between liver and gastrointestinal tract concentration levels showed time-dependent linear correlation and a doubling within 1.5 h.

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Moldes-Anaya Angel S., Fonnum Frode, Eriksen Gunnar S., Rundberget Thomas, Walaas S. Ivar, Wigestrand Mattis B.. In vitro neuropharmacological evaluation of penitrem-induced tremorgenic syndromes: importance of the GABAergic system. Neurochemistry international. 2011;59:1074–1081. 21924313 

The effects of the fungal neurotoxin penitrem A on the GABAergic and glutamatergic systems in rat brain were evaluated. Penitrem A inhibited binding of the GABA(A)-receptor ligand [³H]TBOB to rat forebrain and cerebellar membrane preparations with IC₅₀ (half maximal inhibitory concentration) values of 11 and 9 μM, respectively. Furthermore, penitrem A caused a concentration-dependent increase of [³H]flunitrazepam and [³H]muscimol binding in rat forebrain, but not in cerebellar preparations. The stimulation of [³H]flunitrazepam binding by penitrem A was abolished by the addition of GABA. In cerebellar preparations, a different pharmacological profile was found, with penitrem A allosterically inhibiting [³H]TBOB binding by interacting with a bicuculline-sensitive site. Moreover, penitrem A inhibited the high affinity uptake of GABA and glutamate into cerebellar synaptosomes with IC₅₀ values of 20 and 47 μM, respectively. The toxin showed no effect on NMDA or AMPA glutamate receptor binding. In conclusion, our results suggest that penitrem A exerts region-specific effects in the brain, leading to positive modulation of GABA(A)-receptor function in forebrain. Conversely, penitrem A may act as a bicuculline-like convulsant in cerebellum.

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Lu Hai-Xia X., Levis Hannah, Liu Yong, Parker Terry. Organotypic slices culture model for cerebellar ataxia: potential use to study Purkinje cell induction from neural stem cells. Brain research bulletin. 2011;84:169–173. PMID: 21145378 

In order to establish an in vitro model of cerebellarataxia for evaluating the potential of neural stem cells (NSC) to repair damage caused by cerebella disorders, organotypic slices of cerebellum were cultured using Stoppini method. Penitrem A (4 mg/kg) was administered intraperitoneally (IP) to postnatal day 6 neonatal Wistar rats to produce Purkinje cell deficient slices suitable to host transplanted NSCs. Analysis indicated that cerebellar cytoarchitecture was well preserved and Purkinje cells survived for over three weeks with fewer Purkinje cells in slices cultured from Penitrem A treated neonates compared to controls.

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Ohno Akitoshi, Ohya Susumu, Yamamura Hisao, Imaizumi Yuji. Gender difference in BK channel expression in amygdala complex of rat brain. Biochemical and biophysical research communications. 2009;378:867–871. PMID: 19070588 

The expression of large-conductance Ca(2+)-activated K(+) (BKchannel protein in amygdala complex was higher in adult (8-10 weeks old) male rats than in female. Castration at 4-6 weeks old significantly reduced BK channel expression in amygdala to the level similar to that in female. Immunocytochemical analyses of pyramidal-like neurons isolated from amygdala revealed that somas with relatively large size were highly immunoreactive to both anti-androgen receptor (AR) and anti-BK channel antibodies, while those with smaller size were not. The double-immunopositive neurons were dominant (60%) among pyramidal-like neurons isolated from amygdala of male rats but rare among those from female. The membrane current sensitive to penitrem A, a BK channel blocker, was the major K(+) current component in large neurons and showed higher current-density than that in smaller ones. These results suggest the gender-dependent cell population expressing BK channels in amygdala complex and its up-regulation by AR stimulation.

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Lu Hai-xia X., Levis Hannah, Melhem Nabil, Parker Terry. Toxin-produced Purkinje cell death: a model for neural stem cell transplantation studies. Brain research. 2008;1207:207–213. PMID: 18374311 

Purkinje cell loss is the hallmark of the cerebellar ataxias. Here the fungal neurotoxin Penitrem A was used to create partially Purkinje-cell-deficient cerebella in neonate and young adult rats suitable for use in neural stem cell transplantation studies. I.p. administration of Penitrem A to P3, P6 and 11-week old rats caused noticeable tremor in all treated animals that lasted between 1 and 3 days and was more immediate in the rat pups than in the 11-week old rats. Quantification of cresyl violet stained sections showed that Purkinje cells were preferentially lost in the cerebellar vermis and specifically in folia VI to IX (P<0.001-0.05). No change occurred in Purkinje cell number in folia I-III and folium X. These results were confirmed by the loss of calbindin binding cells in the Purkinje cell layer and the appearance of enlarged vacuolated mitochondria. The results of the present study show that the Penitrem A can remove Purkinje cells in the immature rat cerebellum and thus provide a potential model to study the micro-environmental cues in vivo for the differentiation of Purkinje cells from transplanted and/or intrinsic neural stem cells.

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Brooke Ruth E., Moores Thomas S., Morris Neil P., Parson Simon H., Deuchars Jim. Kv3 voltage-gated potassium channels regulate neurotransmitter release from mouse motor nerve terminals. The European journal of neuroscience. 2004;20:3313–3321. PMID: 15610163 

Voltage-gated potassium (Kv) channels are critical to regulation of neurotransmitter release throughout the nervous system but the roles and identity of the subtypes involved remain unclear. Here we show that Kv3 channels regulate transmitter release at the mouse neuromuscular junction (NMJ). Light- and electron-microscopic immunohistochemistry revealed Kv3.3 and Kv3.4 subunits within all motor nerve terminals of muscles examined [transversus abdominus, lumbrical and flexor digitorum brevis (FDB)]. Tetraethylammonium (TEA) applied at low concentrations (0.05-0.5 mM), which blocks only a few known potassium channels including Kv3channels, did not affect muscle fibre resting potential but significantly increased the amplitude of all EPPs tested. Significantly, this effect of TEA was still observed in the presence of the large-conductance calcium-activated potassium channel blockers iberiotoxin (25-150 nM) and Penitrem A (100 nM), suggesting a selective action on Kv3 subunits.

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Cavanagh J. B., Holton J. L., Nolan C. C., Ray D. E., Naik J. T., Mantle P. G.. The effects of the tremorgenic mycotoxin penitrem A on the rat cerebellum. Veterinary pathology. 1998;35:53–63. PMID: 9545135 

Within 10 minutes of intraperitoneal injection of penitrem A (3 mg/kg), rats develop severe generalized tremors and ataxia that persist for up to 48 hours. These are accompanied by a three- to fourfold increase in cerebellar cortical blood flow. Mitochondrial swelling occurs in cerebellar stellate and basket cells within 30 minutes of dosing and persists for more than 12 hours without leading to cell death. From 2 hours, Purkinje cell dendrites show early cytoplasmic condensation accompanied by fine vacuolation of smooth endoplasmic reticulum and enlargement of perikaryal mitochondria. From 6 hours, many Purkinje cells develop intense cytoplasmic condensation with eosinophilia that resembles “ischemic cell change,” and from 12 hours, many other Purkinje cells show marked watery swelling. Astrocytes begin to swell from 0.5 hours after injection and show hypertrophy of organelles from 6 hours. Also from 6 hours onward, discrete foci of necrosis appear in the granule cell layer, while permeability of overlying meningeal vessels to horseradish peroxidase becomes evident at 8 hours. All changes are more severe in vermis and paravermis. Despite widespread loss of Purkinje cells, the animals’ behavior becomes almost normal within a week. While tremor occurs with doses of 1.5 and 0.5 mg/kg, cellular damage is minimal. The tremor mechanism differs from that of harmaline since destruction of inferior olivary nuclei abolishes neither the tremor response to penitrem A nor the cellular damage. No morphological changes are found in other brain regions. The affinities of penitrem A for high-conductance calcium-dependent potassium channels and for gamma-aminobutyric acid receptors with the probability of resultant excitotoxity are considered to be important underlying factors for these changes.

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Deschaux O., Bizot J. C.. Effects of penitrem A on ratʼs performances in passive avoidance and Morris water maze tests. Mycopathologia. 1997;138:99–104. PMID: 9433810

Intraperitoneal administration of the mycotoxin penitrem A 30 min before a training session in passive avoidance task, impaired performance of rats subjected to a test-session 24 h after. This effect was not antagonised by pretraining administration of physostigmine or bicuculline. Administration of penitrem A 20 min before a training session or 30 min before a test-session did not impair performance. In the Morris watermaze, doses of penitrem A that induces slight to moderate tremors, but not a lower dose, disrupted place learning. These results suggest that penitrem A disrupts the processes that take place at the time of acquisition, but not those just after acquisition, and does not alter the restitution of information. This effect would not be related to a decrease of cholinergic neurotransmission nor to a stimulation of GABA A receptors. Nevertheless, it could not be totally excluded that the performance impairments induced by penitrem A would be secondary to a motor disruption.

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Breton P., Bizot J. C., Buee J., De La Manche I.. Brain neurotoxicity of Penitrem A: electrophysiological, behavioral and histopathological study. Toxicon. 1998;36:645–655. PMID: 9643478

The neurotoxicity of Penitrem A (PA) in rats was assessed against neurophysiological, behavioral and histopathological parameters. Animals were acutely given intracerebroventricular (22-45 mg) or intraperitoneal injections (0.5-1.5 mg/kg) of PA. A typical trembling syndrome associated with PA was always noted. Depending on the dose administered, animals may convulse and eventually die (1-1.5 mg/kg). PA-induced transient alterations of the EEG involving an increase in the frequency and voltage of electrical activity recorded from the cerebral cortex. Hippocampal activity was not modified and some pathologic activities may be recorded at the thalamus. Generally these EEG alterations disappeared at d 3 after the injection and the animals progressively recovered. However in the most severe cases, neuromotor disturbances were maintained at d 7 (rotarod test). Coronal sections of the brain at the striatal, thalamic, hippocampal and pons levels mainly revealed that PA was able to induce dose related injuries in the cerebellum with massive degeneration of Purkinje cells and a significant vacuolization within the molecular layer. The neurotoxic mechanism remains unclear. Action of the mycotoxin on the cerebello-thalamo-cortical tract is discussed.

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Scuteri M., Miguel M. A., Blanco Viera J., Banchero E.. Tremorgenic mycotoxins produced by strains of Penicillium spp. isolated from toxic Poa huecu parodi. Mycopathologia. 1992;120:177–182. PMID: 1494361

Seventeen strains of Penicillium spp. have been isolated from Poa huecu Parodi from the Zapala zone, exhibiting toxicity to sheet. The following strains have been identified: P. crustosum, cyclopium, notatum, palitans, puberulum, verrucosum, viridicatum and Penicillium spp. The toxigenic capacity of the strains was studied after growing them under suitable conditions. The effect of these mycotoxins was studied in mice. Neurological symptoms characteristic of the intoxication by tremorgenic toxins and similar to those observed in sheep suffering from ‘huecu‘s disease’ were observed. The possible role of these toxins as causative agents of ‘huecu‘s disease’ is discussed.

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Bradford H. F., Norris P. J., Smith C. C.. Changes in transmitter release patterns in vitro induced by tremorgenic mycotoxins. Journal of environmental pathology, toxicology and oncology. 1990;10:17–30. PMID: 1977902

The neurochemical effects of the tremorgenic mycotoxins Verruculogen and Penitrem A, which produce a neurotoxic syndrome characterized by sustained tremors, were studied using sheep and rat synaptosomes. Penitrem A (400 mg mycelium/kg) increased the spontaneous release of endogenous glutamate, GABA, and aspartate by 213%, 455%, and 227%, respectively, from cerebrocortical synaptosomes. The spontaneous release of the transmitter amino acids or other amino acids was not increased by the tremorgens in spinal cord/medullary synaptosomes. Penitrem A pretreatment reduced the Veratrine (75 microM) stimulated release of glutamate, aspartate and GABA from cerebrocortical synaptosomes by 33%, 46%, and 11% respectively, and the stimulated release of glycine and GABA from spinal cord/medulla synaptosomes by 67% and 32%, respectively. Penitrem A pretreatment increased the spontaneous release of aspartate, glutamate and GABA by 68%, 62%, and 100%, respectively, from sheep corpus striatum synaptosomes but did not alter the synthesis and release of dopamine in this tissue.

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Norris P. J., Smith C. C., De Belleroche J., et al. Actions of tremorgenic fungal toxins on neurotransmitter release. Journal of neurochemistry. 1980;34:33–42. PMID: 6108984

The neurochemical effects of the tremorgenic mycotoxins Verruculogen and Penitrem A, which produce a neurotoxic syndrome characterised by sustained tremors, were studied using sheep and rat synaptosomes. The toxins were administered in vivo, either by chronic feeding (sheep) or intraperitoneal injection 45 min prior to killing (rat), and synaptosomes were subsequently prepared from cerebrocortical and spinal cord/medullary regions of rat, and corpus striatum of sheep. Penitrem A (400 mg mycelium/kg) increased the spontaneous release of endogenous glutamate, GABA (gamma-aminobutyric acid), and aspartate by 213%, 455%, and 277%, respectively, from cerebrocortical synaptosomes. The spontaneous release of the transmitter amino acids or other amino acids was not increased by the tremorgens in spinal cord/medullary synaptosomes. Penitrem A pretreatment reduced the veratrine (75 microM) stimulated release of glutamate, aspartate, and GABA from cerebrocortical synaptosomes by 33%, 46%, and 11%, respectively, and the stimulated release of glycine and GABA from spinal cord/medulla synaptosomes by 67% and 32% respectively. Penitrem A pretreatment increased the spontaneous release of aspartate, glutamate, and GABA by 68%, 62%, and 100%, respectively, from sheep corpus striatum synaptosomes but did not alter the synthesis and release of dopamine in this tissue.

Sobotka T. J., Brodie R. E., Spaid S. L.. Neurobehavioral studies of tremorgenic mycotoxins verruculogen and penitrem A. Pharmacology. 1978;16:287–294. PMID: 643898 

Verruculogen (V) and penitrem A (PA) represent a group of toxic secondary metabolites of mold contaminants of foodstuffs known as ‘tremorgenic mycotoxins‘, which produce a unique neurotoxic syndrome characterized by sustained tremors, limb weakness, ataxia and convulsions. In the present study the intraperitoneal median tremogenic dose in mice for V was found to be 0.92 mg/kg and that for PA, 0.19 mg/kg. Behavioral and neurochemical parameters were assessed following acute exposure to varying neurotoxic and subneurotoxic doses of these mycotoxins. Measures of spontaneous motor activity (photoactometer) and exploratory reflex behaviors (open field) were markedly depressed by both V and PA. Notably, at dose levels of V or PA that were not accompanied by any overt signs of neurotoxicity, significant neuromotor depression was still observed. Acquisition of a conditioned avoidance shuttle response was disrupted, but only at a high neurotoxic dose level of V. Neurochemical analyses revealed no clear catecholaminergic or cholinergic involvement in the neurotoxic syndrome of eigher V or PA.

 

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