Inhibition of arachidonate 15-lipoxygenase prevents 4-hydroxynonenal-induced protein damage in male germ cells

Elizabeth G. Bromfield, Bettina P. Mihalas, Matthew D. Dun, R. John Aitken, Eileen A. McLaughlin, Jessica L.H. Walters, Brett Nixon

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Lipid peroxidation products, such as 4-hydroxynonenal (4HNE), are causative agents responsible for extensive protein damage within the male and female germlines. Recently, we have demonstrated that 4HNE production can initiate the proteolytic degradation of the molecular chaperone Heat Shock Protein A2 (HSPA2) in male germ cells. These events may be partially responsible for HSPA2 deficiency in the spermatozoa of patients that repeatedly fail in vitro fertilization. Given this, mechanisms that limit the production of 4HNE will be highly advantageous for the preservation of male fertility. The propagation of 4HNE in somatic cells has been linked to the enzymatic actions of arachidonate 15-lipoxygenase (ALOX15), a member of the lipoxygenase family of proteins. In view of this association, this study sought to explore ALOX15 as a physiological target to manipulate the levels of 4HNE produced in the male germline. Herein, we have demonstrated that ALOX15 is markedly upregulated in response to oxidative stress in round spermatids and the GC-2 cell line. Pharmacological inhibition of ALOX15 in GC-2 cells resulted in a significant reduction in both mitochondrial and cytoplasmic reactive oxygen species, as well as a dramatic reduction in 4HNE. Importantly, the reduced bioavailability of this aldehyde appears to confer positive downstream effects to its target proteins such that HSPA2 could be protected from damage by 4HNE. Taken together, these results suggest that the actions of ALOX15 are intimately tied to the production of 4HNE. Thus, the ALOX15 protein may be a promising new target for the mitigation of germline oxidative stress.

Original languageEnglish
Article numberiox005
Pages (from-to)598-609
Number of pages12
JournalBiology of Reproduction
Volume96
Issue number3
Early online date2 Feb 2017
DOIs
Publication statusPublished - 1 Mar 2017
Externally publishedYes

Fingerprint

Arachidonate 15-Lipoxygenase
Germ Cells
Proteins
Heat-Shock Proteins
varespladib methyl
Oxidative Stress
Fertility Preservation
Protein Deficiency
Molecular Chaperones
Spermatids
Lipoxygenase
Inhibition (Psychology)
4-hydroxy-2-nonenal
Fertilization in Vitro
Aldehydes
Lipid Peroxidation
Biological Availability
Spermatozoa
Reactive Oxygen Species
Pharmacology

Cite this

Bromfield, E. G., Mihalas, B. P., Dun, M. D., John Aitken, R., McLaughlin, E. A., Walters, J. L. H., & Nixon, B. (2017). Inhibition of arachidonate 15-lipoxygenase prevents 4-hydroxynonenal-induced protein damage in male germ cells. Biology of Reproduction, 96(3), 598-609. [iox005]. https://doi.org/10.1093/biolre/iox005
Bromfield, Elizabeth G. ; Mihalas, Bettina P. ; Dun, Matthew D. ; John Aitken, R. ; McLaughlin, Eileen A. ; Walters, Jessica L.H. ; Nixon, Brett. / Inhibition of arachidonate 15-lipoxygenase prevents 4-hydroxynonenal-induced protein damage in male germ cells. In: Biology of Reproduction. 2017 ; Vol. 96, No. 3. pp. 598-609.
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abstract = "Lipid peroxidation products, such as 4-hydroxynonenal (4HNE), are causative agents responsible for extensive protein damage within the male and female germlines. Recently, we have demonstrated that 4HNE production can initiate the proteolytic degradation of the molecular chaperone Heat Shock Protein A2 (HSPA2) in male germ cells. These events may be partially responsible for HSPA2 deficiency in the spermatozoa of patients that repeatedly fail in vitro fertilization. Given this, mechanisms that limit the production of 4HNE will be highly advantageous for the preservation of male fertility. The propagation of 4HNE in somatic cells has been linked to the enzymatic actions of arachidonate 15-lipoxygenase (ALOX15), a member of the lipoxygenase family of proteins. In view of this association, this study sought to explore ALOX15 as a physiological target to manipulate the levels of 4HNE produced in the male germline. Herein, we have demonstrated that ALOX15 is markedly upregulated in response to oxidative stress in round spermatids and the GC-2 cell line. Pharmacological inhibition of ALOX15 in GC-2 cells resulted in a significant reduction in both mitochondrial and cytoplasmic reactive oxygen species, as well as a dramatic reduction in 4HNE. Importantly, the reduced bioavailability of this aldehyde appears to confer positive downstream effects to its target proteins such that HSPA2 could be protected from damage by 4HNE. Taken together, these results suggest that the actions of ALOX15 are intimately tied to the production of 4HNE. Thus, the ALOX15 protein may be a promising new target for the mitigation of germline oxidative stress.",
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Bromfield, EG, Mihalas, BP, Dun, MD, John Aitken, R, McLaughlin, EA, Walters, JLH & Nixon, B 2017, 'Inhibition of arachidonate 15-lipoxygenase prevents 4-hydroxynonenal-induced protein damage in male germ cells', Biology of Reproduction, vol. 96, no. 3, iox005, pp. 598-609. https://doi.org/10.1093/biolre/iox005

Inhibition of arachidonate 15-lipoxygenase prevents 4-hydroxynonenal-induced protein damage in male germ cells. / Bromfield, Elizabeth G.; Mihalas, Bettina P.; Dun, Matthew D.; John Aitken, R.; McLaughlin, Eileen A.; Walters, Jessica L.H.; Nixon, Brett.

In: Biology of Reproduction, Vol. 96, No. 3, iox005, 01.03.2017, p. 598-609.

Research output: Contribution to journalArticle

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T1 - Inhibition of arachidonate 15-lipoxygenase prevents 4-hydroxynonenal-induced protein damage in male germ cells

AU - Bromfield, Elizabeth G.

AU - Mihalas, Bettina P.

AU - Dun, Matthew D.

AU - John Aitken, R.

AU - McLaughlin, Eileen A.

AU - Walters, Jessica L.H.

AU - Nixon, Brett

PY - 2017/3/1

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N2 - Lipid peroxidation products, such as 4-hydroxynonenal (4HNE), are causative agents responsible for extensive protein damage within the male and female germlines. Recently, we have demonstrated that 4HNE production can initiate the proteolytic degradation of the molecular chaperone Heat Shock Protein A2 (HSPA2) in male germ cells. These events may be partially responsible for HSPA2 deficiency in the spermatozoa of patients that repeatedly fail in vitro fertilization. Given this, mechanisms that limit the production of 4HNE will be highly advantageous for the preservation of male fertility. The propagation of 4HNE in somatic cells has been linked to the enzymatic actions of arachidonate 15-lipoxygenase (ALOX15), a member of the lipoxygenase family of proteins. In view of this association, this study sought to explore ALOX15 as a physiological target to manipulate the levels of 4HNE produced in the male germline. Herein, we have demonstrated that ALOX15 is markedly upregulated in response to oxidative stress in round spermatids and the GC-2 cell line. Pharmacological inhibition of ALOX15 in GC-2 cells resulted in a significant reduction in both mitochondrial and cytoplasmic reactive oxygen species, as well as a dramatic reduction in 4HNE. Importantly, the reduced bioavailability of this aldehyde appears to confer positive downstream effects to its target proteins such that HSPA2 could be protected from damage by 4HNE. Taken together, these results suggest that the actions of ALOX15 are intimately tied to the production of 4HNE. Thus, the ALOX15 protein may be a promising new target for the mitigation of germline oxidative stress.

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Bromfield EG, Mihalas BP, Dun MD, John Aitken R, McLaughlin EA, Walters JLH et al. Inhibition of arachidonate 15-lipoxygenase prevents 4-hydroxynonenal-induced protein damage in male germ cells. Biology of Reproduction. 2017 Mar 1;96(3):598-609. iox005. https://doi.org/10.1093/biolre/iox005