Toxic Concentrations of Glutamate Induce Ferroptosis without Affecting Mitochondrial Respiration in Rodent Brain Slice Cultures

May combine two claims.

Rosiglitazone is an inhibitor of acyl-CoA synthetase long-chain family member 4 (ACSL4), an enzyme that catalyzes the esterification of arachidonic and adrenic acids preparing them for synthesis of phospholipids, enriching membranes with oxidizable polyunsaturated fatty acids and thereby promoting the LPO that drives ferroptosis.

“In addition, we tested rosiglitazone, an inhibitor of acyl-CoA synthetase long-chain family member 4 (ACSL4), an enzyme that catalyzes the esterification of arachidonic and adrenic acids preparing them for synthesis of phospholipids, enriching membranes with oxidizable polyunsaturated fatty acids and thereby promoting the LPO that drives ferroptosis (Ding et al., 2023).”

• cites Ding et al., 2023 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

A part of glutamate can be taken up by mitochondria, where it is metabolized to 2-oxoglutarate, contributing to ATP synthesis.

“A part of the glutamate can be taken up by the mitochondria, where it is metabolized to 2-oxoglutarate further contributing to the synthesis of ATP (Tang and Zucker, 1997; Verstreken et al., 2005), thereby removing it from the glutamine–glutamate cycle and reducing its toxicity (Vaglio-Garro et al., 2024b).”

• cites Tang and Zucker, 1997 sourceCould not verify

  The Discussion describes mitochondrial calcium transport and says ATP production is unlikely to explain post-tetanic potentiation. Searches for glutamate uptake, 2-oxoglutarate, and glutamate-linked ATP synthesis found no passage in our copy addressing the attributed metabolic pathway. (LLM, conf=0.72)

  Experimental — accuracy not yet measured
• cites Verstreken et al., 2005 sourcePartly supported

  The Discussion reports ATP generation by synaptic mitochondria, supporting the ATP-production part of the claim. Our copy does not say that mitochondria take up glutamate or metabolize it to 2-oxoglutarate (searched both terms and a paraphrase), so it does not establish the claimed route to ATP synthesis. (LLM, conf=0.83)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Memantine, an NMDA receptor antagonist, protected neuronal primary cells from glutamate-induced death.

“It has been shown that memantine, an N-methyl-d-aspartate (NMDA) receptor antagonist, protected neuronal primary cells from glutamate-induced death (Weidinger et al., 2023).”

• cites Weidinger et al., 2023 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Acute brain slices (ABSs) have been developed and used for investigating the structural and functional characteristics of the adult brain.

“The acute brain slices (ABSs) have been developed and used for investigating the structural and functional characteristics of the adult brain (Eguchi et al., 2020).”

• cites Eguchi et al., 2020 sourcePartly supported

  The Introduction says acute brain slices have been developed and extensively used to investigate the structural and functional characteristics of neuronal circuits. It does not specify the adult brain; the Animals section says this study generally used mice aged 4–6 weeks, so the adult-brain qualifier is not established by our copy. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Ferroptosis is associated with neural disorders.

“Recently iron-mediated lipid peroxidation (LPO) reactions have been recognized as the major contributor to a novel mechanism of programmed cell death known as ferroptosis (Stoyanovsky et al., 2019), which has been also associated with neural disorders (Li et al., 2019; Wang, T. et al., 2022; Wang, Z. L. et al., 2022).”

• cites Li et al., 2019Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites Wang, T. et al., 2022 sourceSupported
• cites Wang, Z. L. et al., 2022 sourceSupported

Classified as probably not a substantial claim. May combine two claims.

Glutamate-induced cell death is associated with mitochondrial dysfunction, Ca2+ accumulation, mitochondrial permeability transition pore opening, and functional collapse of mitochondria.

“Glutamate-induced cell death is associated with mitochondrial dysfunction, associated with accumulation of Ca2+, activating mitochondrial permeability transition pore opening and causing functional collapse of mitochondria (for review, see Szabó and Zoratti, 1992; Bonora et al., 2020).”

• cites Szabó and Zoratti, 1992Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites Bonora et al., 2020 sourcePartly supported

  Section 1.1, “Mitochondrial Routes of Cell Death” (page not available), says pathological mitochondrial Ca2+ overload triggers permeability transition and pore opening, followed by collapse of mitochondrial potential and related activities that can lead to cell death. It does not connect this sequence specifically to glutamate-induced cell death. (LLM, conf=0.85)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

In primary neuronal cultures, glutamate toxicity primarily targets neurons, causing a marked reduction in tyrosine hydroxylase-positive neurons and neurite degeneration.

“In primary neuronal cultures, it has been shown that glutamate toxicity primarily targets neurons, as evidenced by a marked reduction in the number of tyrosine hydroxylase-positive neurons together with neurite degeneration (Weidinger et al., 2023).”

• cites Weidinger et al., 2023 sourcePartly supported

  The Results (Fig. 6; page not provided) report that glutamate reduced the number of tyrosine hydroxylase-positive neurons and shortened their neurites in primary murine mesencephalic cultures. The paper does not establish that glutamate primarily targets neurons rather than other cell types; that comparative part of the claim is unsupported. (LLM, conf=0.83)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Restoration of OGDHC activity attenuated glutamate-induced Ca2+ accumulation and neuronal cell death.

“In a previous study, restoration of the mitochondrial enzyme complex oxoglutarate dehydrogenase complex (OGDHC) activity attenuated glutamate-induced Ca2+ accumulation and neuronal cell death (Weidinger et al., 2023), supporting a mechanistic link between glutamate metabolism, excitotoxic Ca2+ signaling, and neuronal survival.”

• cites Weidinger et al., 2023 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

If pyruvate is not sufficiently present in mitochondria, glutamate can be used for energy instead.

“Glutamate is reserved for signal transduction and is normally not used for energy purposes; however, if pyruvate is not sufficiently present in the mitochondria, then glutamate can be used instead (Sonnay et al., 2017).”

• cites Sonnay et al., 2017 sourcePartly supported

  The ‘Brain glucose uptake and metabolism’ section says glutamate can re-enter the TCA cycle and be oxidized for energy. It does not say this happens specifically when mitochondrial pyruvate is insufficient, or that glutamate is normally reserved exclusively for signaling. Its discussion of low acetyl-CoA instead describes pyruvate recycling. (LLM, conf=0.87)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Endogenous NO serves as a posttranslational regulator of OGDHC and may affect toxic effects of glutamate.

“Previous studies have shown that elevated levels of glutamate may upregulate the activity of OGDHC and that endogenous NO serves as a posttranslational regulator of OGDHC and may affect toxic effects of glutamate (Weidinger et al., 2023).”

• cites Weidinger et al., 2023 sourcePartly supported

  The Results (page unavailable) report that glutamate treatment increased OGDHC activity and that NO inhibited it and increased glutamate-induced cell damage. The Discussion (page unavailable) links increased NO production to reduced OGDHC activity and toxic extracellular glutamate. Our copy does not establish that NO acts through a posttranslational modification of OGDHC, so that specific characterization is unsupported. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

The pathway of glutamate into the TCA cycle mainly occurs in neurons.

“This pathway mainly occurs in neurons (Dobolyi et al., 2020; Zhong et al., 2022), and OGDHC is a limiting step in this process (Mkrtchyan et al., 2018).”

• cites Dobolyi et al., 2020 sourcePartly supported

  The Results section proposes that neuronal mitochondria perform the conventional TCA cycle, while glia in the adult human cortex process glutamate through other routes. The Discussion says the apparent lack of KGDHC in human glia may prevent glutamate catabolism through the TCA cycle. This supports the neuron–glia distinction, but the source frames it as a proposal about adult human cortex, not an established general rule for all neurons. (LLM, conf=0.83)

  Experimental — accuracy not yet measured
• cites Zhong et al., 2022 sourceNot checked — no full text of the source

May combine two claims.

BSC preserves anatomically brain regions and is highly sensitive to oxygen and glucose changes, allowing recapitulation of pathological changes in energy metabolism.

“Because they preserve their anatomically brain regions, it is also possible to recapitulate pathological changes in energy metabolism because the slices are highly sensitive to oxygen and glucose changes (Underwood et al., 2020).”

• cites Underwood et al., 2020 sourcePartly supported

  The Results on tissue preparation say brain sections are highly vulnerable to oxygen and glucose deprivation, and the Discussion says tissue punches retain their surrounding cellular milieu and permit measurements in anatomically defined regions. The Results on mild fluid percussion injury also show region-specific changes in mitochondrial respiration. The paper does not show that changing oxygen or glucose in slices recapitulates pathological changes in energy metabolism, so the claimed causal link is unsupported. (LLM, conf=0.83)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Elevated levels of glutamate may upregulate the activity of OGDHC.

“Previous studies have shown that elevated levels of glutamate may upregulate the activity of OGDHC and that endogenous NO serves as a posttranslational regulator of OGDHC and may affect toxic effects of glutamate (Weidinger et al., 2023).”

• cites Weidinger et al., 2023 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Toxic levels of glutamate induce morphological changes in neurons, manifesting in deformed dendrites and expanded rounded neurons.

“These toxic levels of glutamate induce morphological changes in neurons, manifesting in deformed dendrites and expanded rounded neurons (Reynolds’ and Hastings, 1995; Jellinger, 1999; Adamchik et al., 2000; Robert et al., 2002; Youdim et al., 2002; García et al., 2005), showing degenerative cell (Kang et al., 2020; Erkartal et al., 2025) and neuronal death (Weidinger et al., 2023).”

• cites Reynolds’ and Hastings, 1995 sourcePartly supported

  The Results (page not available in our copy) note some neuronal swelling after glutamate addition, supporting a change in cell shape. They do not describe deformed dendrites or expanded, rounded neurons. The small round cells mentioned elsewhere in the Results did not respond to glutamate. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites Jellinger, 1999 sourceNot checked — no full text of the source
• cites Adamchik et al., 2000 sourceNot checked — no full text of the source

May combine two claims.

Toxic levels of glutamate cause degenerative cell and neuronal death.

“These toxic levels of glutamate induce morphological changes in neurons, manifesting in deformed dendrites and expanded rounded neurons (Reynolds’ and Hastings, 1995; Jellinger, 1999; Adamchik et al., 2000; Robert et al., 2002; Youdim et al., 2002; García et al., 2005), showing degenerative cell (Kang et al., 2020; Erkartal et al., 2025) and neuronal death (Weidinger et al., 2023).”

• cites Kang et al., 2020 sourceNot checked yet

  Checked before — its earlier verdict is held back until a new check runs.

• cites Erkartal et al., 2025 sourcePartly supported

  The Discussion reports greater neuronal degeneration in MSG-treated rats than in controls, supporting the degenerative-change portion attributed to Erkartal et al. The paper studies administered MSG in rats, not measured toxic glutamate levels generally. The citing sentence attaches a different citation to neuronal death. (LLM, conf=0.82)

  Experimental — accuracy not yet measured
• cites Weidinger et al., 2023 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

In neurons, additional intracellular iron pools may also contribute to ferroptosis induction.

“However, in neurons, additional intracellular iron pools may also contribute to ferroptosis induction (Chen et al., 2026).”

• cites Chen et al., 2026 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Deferoxamine has been reported to inhibit ferroptosis in several experimental disease models.

“has been reported to inhibit ferroptosis in several experimental disease models (Ma et al., 2016; Bruni et al., 2018).”

• cites Ma et al., 2016Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites Bruni et al., 2018 sourcePartly supported

  The Results section reports that deferoxamine (DFO) prevented erastin-induced ferroptotic cell death in cultured human islets. It does not establish DFO's effect in several experimental disease models: the Introduction attributes that broader finding to ferrostatins, not DFO. No page numbers are available in our copy. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Deferoxamine is an FDA-approved iron-chelating agent.

“deferoxamine is an FDA-approved iron-chelating agent (Yao et al., 2019)”

• cites Yao et al., 2019 sourceSupported

Classified as probably not a substantial claim.

Mitochondrial dysfunction is characteristic of the excitotoxic pathway.

“This aligns with the knowledge that mitochondrial dysfunction is characteristic of the excitotoxic pathway (Nicholls and Budd, 1998; Votyakova and Reynolds, 2005), whereas ferroptosis may operate through damage other than (Wang et al., 2015) mitochondrial cellular membranes, such as the cytoplasmic membrane (Vaglio-Garro et al., 2024b).”

• cites Nicholls and Budd, 1998 sourceSupported
• cites Votyakova and Reynolds, 2005 sourceNot checked — no full text of the source

May combine two claims.

Diffusion of glutamate to cells in BSC is slower than in cell cultures, which means BSC require higher concentrations of glutamate to achieve toxic effects.

“First, the diffusion of glutamate to the cells in BSC is slower than in cell cultures, which means BSC require higher concentrations of glutamate to achieve toxic effects (Humpel, 2015).”

• cites Humpel, 2015 sourcePartly supported

  In “Applications using organotypic brain slices” (page unavailable), Humpel reports that slices need markedly higher doses of a toxic stimulus than primary cells. The passage does not identify glutamate as that stimulus or say that slower glutamate diffusion explains the difference. (LLM, conf=0.87)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Recent evidence suggests that the lysosomal iron pool may play a predominant role in ferroptosis.

“Recent evidence suggests that the lysosomal iron pool may play a predominant role in ferroptosis (Cañeque et al., 2025).”

• cites Cañeque et al., 2025 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

There is a body of evidence for the activation of the excitotoxic pathway by glutamate, including in our previous publication.

“Since there is a body of evidence for the activation of the excitotoxic pathway by glutamate, also in our previous publication (Weidinger et al., 2023), we collected more evidence related to the activation of excitotoxic pathway in our model.”

• cites Weidinger et al., 2023 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Normalization of respiration to tissue weight did not substantially alter the results.

“This normalization did not substantially alter the results (Fig. 5A–D).”

May combine two claims.

Neither memantine nor ferrostatin-1 affected LDH release in control samples.

“Neither memantine nor ferrostatin-1 affected LDH release in control samples.”

May combine two claims.

In homogenized brain slices, respiration in glutamate-treated groups showed a nonsignificant trend toward a decrease, evident in ATP-linked respiration with glutamate and succinate, and in uncoupled respiration, but not in routine respiration.

“Interestingly, the respiration in the glutamate-treated groups showed a nonsignificant trend toward a decrease. This trend was evident in ATP-linked respiration with glutamate (Fig. 4C) and succinate (Fig. 4D), as well as in uncoupled respiration (Fig. 4E), but not in routine respiration (Fig. 4B).”

May combine two claims.

In the O2k experiments with homogenized BSC, the lowest levels of leak and ATP-linked respiration were observed in the glutamate and glutamate + memantine groups, whereas in the presence of ferrostatin-1, mean oxygen flux values reached levels above those of the control group for both leak and ATP synthesis-linked respirations.

“Of note, the lowest levels of leak and ATP-linked respiration were observed in the glutamate and glutamate + memantine groups, whereas, in the presence of ferrostatin-1, mean oxygen flux values reached levels above those of the control group for both leak and ATP synthesis-linked respirations.”

May combine two claims.

In brain homogenate of BSC, OGDHC activity and NO levels did not change significantly after memantine pretreatment in control, Mem, and Glut + Mem groups.

“Figure 3 shows that OGDHC activity (Fig. 3A,B) and NO levels (Fig. 3C,D), in brain homogenate of BSC, did not change significantly after memantine pretreatment in control, Mem and Glut + Mem groups; OGDHC activity tended to be higher in the Glut group compared with the Mem group.”

• Also stated in: § Discussion · plain

  “However, as shown in Figure 3, there were no significant changes in OGDHC activity or NO levels, except for a notable increase in OGDHC activity in the glutamate group compared with the memantine group, but not compared with the controls (Fig. 3A).”

May combine two claims.

Treatment with ferrostatin-1 ameliorated some of the changes induced by glutamate, with more neurons exhibiting pale cytoplasm and dark nuclei.

“Treatment with ferrostatin-1 ameliorated some of the changes induced by glutamate (Fig. 7C,F), with more neurons exhibiting pale cytoplasm and dark nuclei.”

May combine two claims.

In neuronal cell cultures, the phenomenon of intensified energy metabolism and shift to ferroptosis was not observed.

“Such phenomenon was not observed in studies on neuronal cell cultures, also including our previous studies.”

May combine two claims.

Ferrostatin-1 significantly increased leak respiration and tended to increase ATP-linked respiration compared with the Glut group.

“Ferrostatin-1 significantly increased leak respiration (Fig. 5A) and tended to increase ATP-linked respiration compared with the Glut group (Fig. 5B).”

• Also stated in: § Discussion · plain

  “Furthermore, ferrostatin-1, but not memantine, significantly increased the rate of ATP synthesis-linked respiration (Fig. 5B) and uncoupled respiration (Fig. 5D) compared with the glutamate group.”

May combine two claims.

In BSC homogenates, for leak respiration and ATP synthesis-linked oxygen flux, mean values were higher for AP-5 + CNQX compared with the Glut group, but differences were not significant.

“We observed in the BSC homogenates, for leak respiration and ATP synthesis-linked oxygen flux, higher mean values for AP-5 + CNQX compared with the Glut group, but the differences were not significant (Extended Data Fig. 5-1C,D).”

May combine two claims.

In nonhomogenized brain slice cultures, glutamate treatment substantially elevated mitochondrial respiration rate, observed in routine respiration, ATP synthesis-linked respiration, and uncoupled respiration, except uncoupled respiration in the Glut group did not differ from control.

“Metabolic analysis of the samples using glutamate exhibited a substantially elevated respiration rate. This increase was observed in routine (Fig. 2C), ATP synthesis-linked respiration (Fig. 2D), and uncoupled respiration (Fig. 2E), except for uncoupled respiration in the Glut group, which did not differ from the control (Fig. 2E).”

• Also stated in: § Discussion · plain

  “However, we observed a drastic increase in routine respiration (Fig. 2C).”

• AssumptionThe authors state this as an assumption: The respiration observed in the brain tissue primarily originates from neuronal mitochondria.

May combine two claims.

Deferoxamine did not provide significant protection against glutamate-induced cell death (LDH release), despite its pronounced inhibitory effect on LPO.

“In contrast, despite its pronounced inhibitory effect on LPO, deferoxamine did not provide significant protection against glutamate-induced cell death (Fig. 9).”

May combine two claims.

Glutamate enters cells and is used as an energy source to increase ATP synthesis.

“Along with Ca2+, glutamate enters the cells, whereas as shown in this study, it is used as an energy source to increase ATP synthesis.”

May combine two claims.

Treatments with ferrostatin-1 or rosiglitazone significantly reduced glutamate-induced cell death (LDH release).

“Consistent with the LPO data, the treatments with ferrostatin-1 or with rosiglitazone significantly reduced glutamate-induced cell death (Fig. 9).”

May combine two claims.

Deferoxamine almost completely inhibited LPO but failed to improve cell survival.

“Unexpectedly, although deferoxamine almost completely inhibited LPO (Fig. 8), it failed to improve cell survival (Fig. 9).”

May combine two claims.

Inhibition of ACSL4 with rosiglitazone significantly reduced both LPO and glutamate-induced cell death.

“Inhibition of ACSL4 with rosiglitazone significantly reduced both LPO and glutamate-induced cell death (Fig. 8).”

May combine two claims.

Ferrostatin-1 markedly suppressed LPO and improved cell survival.

“A similar protective effect was observed with ferrostatin-1, which markedly suppressed LPO and improved cell survival (Fig. 9).”

May combine two claims.

The respiratory burst after glutamate treatment is due to elevated electron flow through the ETS and not due to activation of ATP synthase.

“The respiration was also increased after uncoupler titration (Fig. 2E), suggesting that this respiratory burst is due to elevated electron flow through ETS and not due to activation of ATP synthase.”

• AssumptionThe authors state this as an assumption: The respiration observed in the brain tissue primarily originates from neuronal mitochondria.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

For Seahorse experiments, slices from three to four brains were pooled and randomly assigned to different treatments.

“For the Seahorse experiments, slices from three to four brains were pooled each time and randomly assigned to different treatments.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Under physiological conditions, extracellular glutamate enters the glutamine-glutamate cycle.

“Under physiological conditions, the extracellular glutamate enters the glutamine–glutamate cycle (McKenna et al., 2016; Sonnewald and Schousboe, 2016).”

• cites McKenna et al., 2016 sourceNot checked — no full text of the source
• cites Sonnewald and Schousboe, 2016 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

Astrocytes obtain their energy predominantly from glycolysis.

“In contrast to neurons, astrocytes obtain their energy predominantly from glycolysis (Brooks, 2018).”

• cites Brooks, 2018 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

Ca2+ stimulates the induction of apoptosis via release of cytochrome c and apoptosis inducing factor.

“In addition, Ca2+ stimulates the induction of apoptosis via release of cytochrome c (cyt c) and apoptosis inducing factor (Lemasters et al., 2002).”

• cites Lemasters et al., 2002 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

The animal study protocol was approved by the Institutional Review Board of the Ludwig Boltzmann Institute of Traumatology.

“The animal study protocol was approved by the Institutional Review Board (Tierschutzgremium) of the Ludwig Boltzmann Institute of Traumatology (Certification MA58-821/90) on February 16, 2023.”

May combine two claims.

Elevated extracellular glutamate concentration blocks the cystine/glutamate antiporter, which may induce ferroptosis.

“Since elevated extracellular glutamate concentration blocks this transporter, it may induce ferroptosis.”

• AssumptionRests on an assumption: Glutathione required for GPX activity is synthesized from cystine, which is transported into the cell via the cystine/glutamate antiporter.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Mitochondrial glutamate uptake removes it from the glutamine-glutamate cycle and reduces its toxicity.

“A part of the glutamate can be taken up by the mitochondria, where it is metabolized to 2-oxoglutarate further contributing to the synthesis of ATP (Tang and Zucker, 1997; Verstreken et al., 2005), thereby removing it from the glutamine–glutamate cycle and reducing its toxicity (Vaglio-Garro et al., 2024b).”

• cites Vaglio-Garro et al., 2024bNot matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

Either the rate of the TCA cycle is elevated in the presence of glutamate or the respiration is limited by endogenous concentrations of TCA cycle substrates, which is compensated by increased transport of glutamate into the cells in the Glut groups.

“These results suggest that either the rate of the TCA cycle is elevated in the presence of glutamate or that the respiration is limited by endogenous concentrations TCA cycle substrates, which is compensated by increased transport of glutamate into the cells in the Glut groups.”

Classified as probably not a substantial claim.

Each slice was treated as an independent sample.

“Each slice, regardless of which pup it was obtained from, was treated as an independent sample.”

Probably not a substantial claim: may not pass S3 (load-bearing).

OGDHC is a limiting step in the process of glutamate uptake into the TCA cycle.

“This pathway mainly occurs in neurons (Dobolyi et al., 2020; Zhong et al., 2022), and OGDHC is a limiting step in this process (Mkrtchyan et al., 2018).”

• cites Mkrtchyan et al., 2018 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

BSC provides a more complex and physiologically relevant model compared with traditional dissociated neuronal cultures.

“BSC provides more complex and physiologically relevant model compared with traditional dissociated neuronal cultures (Humpel, 2018).”

• cites Humpel, 2018 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S2 (checkable).

Targeting ferroptosis may offer therapeutic potential in neuronal disorders traditionally attributed to excitotoxic damage.

“These results suggest that targeting ferroptosis may offer therapeutic potential in neuronal disorders traditionally attributed to excitotoxic damage.”

• AssumptionRests on an assumption: Excessive extracellular glutamate is a hallmark of many neurodegenerative and ischemic conditions.

Classified as probably not a substantial claim.

Iron-mediated lipid peroxidation reactions are the major contributor to ferroptosis.

“Recently iron-mediated lipid peroxidation (LPO) reactions have been recognized as the major contributor to a novel mechanism of programmed cell death known as ferroptosis (Stoyanovsky et al., 2019), which has been also associated with neural disorders (Li et al., 2019; Wang, T. et al., 2022; Wang, Z. L. et al., 2022).”

• cites Stoyanovsky et al., 2019Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

Probably not a substantial claim: may not pass S3 (load-bearing).

Brain slice culture (BSC) is an in vitro model that represents the architecture of the brain.

“Brain slice culture (BSC) is an in vitro model in the field of neurosciences that represents the architecture of the brain (Beach et al., 1982; Bahr, 1995; Sieg et al., 1998).”

• cites Beach et al., 1982 sourceNot checked — no full text of the source
• cites Bahr, 1995 sourceNot checked — no full text of the source
• cites Sieg et al., 1998 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

At least three slices were included per experiment.

“At least three slices were included per experiment.”

May combine two claims.

Glutamate exposure causes a slight impairment of mitochondrial function, which is partially restored by ferrostatin-1.

“These data suggest a slight impairment of mitochondrial function following glutamate exposure, which was partially restored by ferrostatin-1.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Elevated intracellular Ca2+ as a consequence of extracellular glutamate concentrations between 2 and 5 μmol/L leads to excitotoxicity.

“Elevated intracellular Ca2+ as consequence of extracellular concentrations of glutamate between 2 and 5 μmol/L leads to excitotoxicity (Mark et al., 2001);”

• cites Mark et al., 2001Not checked — no full text of the source

May combine two claims.

Ferroptosis is triggered by the extracellular Km of 78 μM glutamate, which inhibits the Xc-transporter and later inhibits intracellular GSH synthesis.

“ferroptosis is triggered by the extracellular Km of 78 μM glutamate (Lewerenz et al., 2013) which inhibits the Xc-transporter and later inhibiting the intracellular GSH synthesis.”

• cites Lewerenz et al., 2013Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

This eventually leads to depletion of intracellular cysteine and reduced GSH, thereby triggering ferroptosis.

“This eventually leads to the depletion of intracellular cysteine and reduced GSH, thereby triggering ferroptosis.”

May combine two claims.

Together, these findings strongly support the induction and execution of ferroptosis following glutamate exposure.

“Together, these findings strongly support the induction and execution of ferroptosis following glutamate exposure.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Deferoxamine is a well-established iron chelator that inhibits LPO and ferroptosis in several experimental models of brain diseases.

“We also investigated effect of deferoxamine, a well-established iron chelator, which was shown to inhibit LPO and ferroptosis in several experimental models of brain diseases (Costa et al., 2023).”

• cites Costa et al., 2023 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

Increased ATP production, driven by glutamate influx, meets the energy demands required for Ca2+ sequestration into mitochondria and the endoplasmic reticulum.

“We propose that the increased ATP production, driven by the influx of glutamate, meets the energy demands required for Ca2+ sequestration into mitochondria and the endoplasmic reticulum.”