Abstract
Bax∆2 is a pro-apoptotic protein originally discovered in colon cancer patients with high microsatellite instability. Unlike most pro-apoptotic Bax family members, Bax∆2 mediates cell death through a non-mitochondrial caspase 8-dependent pathway. In the scope of analyzing the distribution of Bax∆2 expression in human tissues, we examined a panel of human brain samples. Here, we report four cerebellar cases in which the subjects had no neurological disorder or disease documented. We found Bax∆2 positive cells scattered in all areas of the cerebellum, but most strikingly concentrated in Purkinje cell bodies and dendrites. Two out the four subjects tested had strong Bax∆2-positive staining in nearly all Purkinje cells; one was mainly negative; and one had various levels of positive staining within the same sample. Further genetic analysis of the Purkinje cell layer, collected by microdissection from two subjects, showed that the samples contained G7 and G9 Bax microsatellite mutations. Both subjects were young and had no diseases reported at the time of death. As the distribution of Bax∆2 is consistent with that known for Baxα, but in a less ubiquitous manner, these results may imply a potential function of Bax∆2 in Purkinje cells.
This is a preview of subscription content, log in via an institution to check access.
References
Atkins JF, Loughran G, Bhatt PR, Firth AE, Baranov PV (2016) Ribosomal frameshifting and transcriptional slippage: from genetic steganography and cryptography to adventitious use. Nucleic Acids Res 44:gkw530. https://doi.org/10.1093/nar/gkw530
Bernard JA, Seidler RD (2014) Moving forward: age effects on the cerebellum underlie cognitive and motor declines. Neurosci Biobehav Rev 42:193–207. https://doi.org/10.1016/J.NEUBIOREV.2014.02.011
Casoni F, Croci L, Cremona O, Hawkes R, Consalez GG (2017) Early Purkinje cell development and the origins of cerebellar patterning. In: Development of the cerebellum from molecular aspects to diseases. Springer, Cham, pp 67–86
Didonna A, Sussman J, Benetti F, Legname G (2012) The role of Bax and caspase-3 in doppel-induced apoptosis of cerebellar granule cells. Prion 6:309–316. https://doi.org/10.4161/pri.20026
Dorszewska J, Adamczewska-Goncerzewicz Z, Szczech J (2004) Apoptotic proteins in the course of aging of central nervous system in the rat. Respir Physiol Neurobiol 139:145–155. https://doi.org/10.1016/J.RESP.2003.10.009
Fan H, Favero M, Vogel MW (2001) Elimination of Bax expression in mice increases cerebellar Purkinje cell numbers but not the number of granule cells. 91:82–91
Garcia I, Crowther AJ, Gama V, Ryan Miller C, Deshmukh M, Gershon TR (2013) Bax deficiency prolongs cerebellar neurogenesis, accelerates medulloblastoma formation and paradoxically increases both malignancy and differentiation. Oncogene 32:2304–2314. https://doi.org/10.1038/onc.2012.248
Haferkamp B, Zhang H, Lin Y, Yeap X, Bunce A, Sharpe J, Xiang J (2012) Bax∆2 is a novel bax isoform unique to microsatellite unstable tumors. J Biol Chem 287:34722–34729. https://doi.org/10.1074/jbc.M112.374785
Haferkamp B, Zhang H, Kissinger S, Wang X, Lin Y, Schultz M, Xiang J (2013) Bax∆2 family alternative splicing salvages Bax microsatellite-frameshift mutations. Genes Cancer 4:501–512. https://doi.org/10.1177/1947601913515906
Hara A, Hirose Y, Wang A, Yoshimi N, Tanaka T, Mori H (1996) Localization of Bax and Bcl-2 proteins, regulators of programmed cell death, in the human central nervous system. Virchows Arch 429–429:249–253. https://doi.org/10.1007/BF00198341
Jirenhed D-A, Rasmussen A, Johansson F, Hesslow G (2017) Learned response sequences in cerebellar Purkinje cells. Proc Natl Acad Sci USA 114:6127–6132. https://doi.org/10.1073/pnas.1621132114
Jung A-R, Kim TW, Rhyu IJ, Kim H, Lee YD, Vinsant S, Oppenheim RW, Sun W (2008) Misplacement of Purkinje cells during postnatal development in Bax knock-out mice: a novel role for programmed cell death in the nervous system? J Neurosci 28:2941–2948. https://doi.org/10.1523/JNEUROSCI.3897-07.2008
Ketteler R (2012) On programmed ribosomal frameshifting: the alternative proteomes. Front Genet 3:242. https://doi.org/10.3389/fgene.2012.00242
Koziol LF, Budding D, Andreasen N, D’Arrigo S, Bulgheroni S, Imamizu H, Ito M, Manto M, Marvel C, Parker K, Pezzulo G, Ramnani N, Riva D, Schmahmann J, Vandervert L, Yamazaki T (2014) Consensus paper: the cerebellum’s role in movement and cognition. Cerebellum 13:151–177. https://doi.org/10.1007/s12311-013-0511-x
Krajewski S, Krajewska M, Shabaik A, Miyashita T, Wang HG, Reed JC (1994) Immunohistochemical determination of in vivo distribution of Bax, a dominant inhibitor of Bcl-2. Am J Pathol 145:1323–1336
Krajewski S, Mai JK, Krajewska M, Sikorska M, Mossakowski MJ, Reed JC (1995) Upregulation of bax protein levels in neurons following cerebral ischemia. J Neurosci 15:6364–6376
Mañas A, Wang S, Nelson A, Li J, Zhao Y, Zhang H, Davis A, Xie B, Maltsev N, Xiang J (2017) The functional domains for Bax∆2 aggregate-mediated caspase 8-dependent cell death. Exp Cell Res 359:342–355. https://doi.org/10.1016/J.YEXCR.2017.08.016
Mañas A, Chen W, Nelson A, Yao Q, Xiang J (2018) Bax∆2 sensitizes colorectal cancer cells to proteasome inhibitor-induced cell death. Biochem Biophys Res Commun 496:18–24. https://doi.org/10.1016/J.BBRC.2017.12.156
Mothersill O, Knee-Zaska C, Donohoe G (2016) Emotion and theory of mind in schizophrenia—investigating the role of the cerebellum. Cerebellum 15:357–368. https://doi.org/10.1007/s12311-015-0696-2
Penault-Llorca F, Bouabdallan R, Devilard E, Charton-Bain M-C, Hassoun J, Birg F, Xerri L (1998) Analysis of BAX expression in human tissues using the anti-BAX, 4F11 monoclonal antibody on paraffin sections. Pathol Res Pract 194:457–464. https://doi.org/10.1016/S0344-0338(98)80114-3
Picard H, Amado I, Mouchet-Mages S, Olie J-P, Krebs M-O (2007) The role of the cerebellum in schizophrenia: an update of clinical, cognitive, and functional evidences. Schizophr Bull 34:155–172. https://doi.org/10.1093/schbul/sbm049
Vogel MW (2002) Cell death, Bcl-2, Bax, and the cerebellum. Cerebellum 1:277–287. https://doi.org/10.1080/147342202320883588
Zhang H, Lin Y, Manas A, Zhao Y, Denning MF, Ma L, Xiang J (2014) BaxDelta2 promotes apoptosis through caspase-8 activation in microsatellite-unstable colon cancer. Mol Cancer Res 12:1225–1232. https://doi.org/10.1158/1541-7786.MCR-14-0162
Zhang H, Tassone C, Lin N, Mañas A, Zhao Y, Xiang J (2015) Detection of Bax microsatellite mutations and BaxDelta2 isoform in human buccal cells. J Cell Sci Ther. https://doi.org/10.4172/2157-7013.S8-002
Acknowledgements
This work was supported by the National Institutes of Health (R15 CA195526).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Mañas, A., Davis, A., Lamerand, S. et al. Detection of pro-apoptotic Bax∆2 proteins in the human cerebellum. Histochem Cell Biol 150, 77–82 (2018). https://doi.org/10.1007/s00418-018-1669-6
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00418-018-1669-6