Multiple functions of precursor BDNF to CNS neurons: Negative regulation of neurite growth, spine formation and cell survival

Hisatsugu Koshimizu, Kazuyuki Kiyosue, Tomoko Hara, Shunsuke Hazama, Shingo Suzuki, Koichi Uegaki, Guhan Nagappan, Eugene Zaitsev, Takatsugu Hirokawa, Yoshiro Tatsu, Akihiko Ogura, Bai Lu, Masami Kojima

Research output: Contribution to journalArticle

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Abstract

Background. Proneurotrophins and mature neurotrophins elicit opposite effects via the p75 neurotrophin receptor (p75NTR) and Trk tyrosine kinase receptors, respectively; however the molecular roles of proneurotrophins in the CNS are not fully understood. Results. Based on two rare single nucleotide polymorphisms (SNPs) of the human brain-derived neurotrophic factor (BDNF) gene, we generated R125M-, R127L- and R125M/R127L-BDNF, which have amino acid substitution(s) near the cleavage site between the pro- and mature-domain of BDNF. Western blot analyses demonstrated that these BDNF variants are poorly cleaved and result in the predominant secretion of proBDNF. Using these cleavage-resistant proBDNF (CR-proBDNF) variants, the molecular and cellular roles of proBDNF on the CNS neurons were examined. First, CR-proBDNF showed normal intracellular distribution and secretion in cultured hippocampal neurons, suggesting that inhibition of proBDNF cleavage does not affect intracellular transportation and secretion of BDNF. Second, we purified recombinant CR-proBDNF and tested its biological effects using cultured CNS neurons. Treatment with CR-proBDNF elicited apoptosis of cultured cerebellar granule neurons (CGNs), while treatment with mature BDNF (matBDNF) promoted cell survival. Third, we examined the effects of CR-proBDNF on neuronal morphology using more than 2-week cultures of basal forebrain cholinergic neurons (BFCNs) and hippocampal neurons. Interestingly, in marked contrast to the action of matBDNF, which increased the number of cholinergic fibers and hippocampal dendritic spines, CR-proBDNF dramatically reduced the number of cholinergic fibers and hippocampal dendritic spines, without affecting the survival of these neurons. Conclusion. These results suggest that proBDNF has distinct functions in different populations of CNS neurons and might be responsible for specific physiological cellular processes in the brain.

Original languageEnglish
Article number27
JournalMolecular brain
Volume2
Issue number1
DOIs
Publication statusPublished - 24-09-2009

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Neurites
Brain-Derived Neurotrophic Factor
Cell Survival
Spine
Neurons
Growth
Cholinergic Fibers
Dendritic Spines
Physiological Phenomena
Nerve Growth Factor Receptor
Cholinergic Neurons
Normal Distribution
Nerve Growth Factors
Receptor Protein-Tyrosine Kinases
Amino Acid Substitution
brain-derived neurotrophic factor precursor
Single Nucleotide Polymorphism
Western Blotting
Apoptosis
Brain

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cellular and Molecular Neuroscience

Cite this

Koshimizu, Hisatsugu ; Kiyosue, Kazuyuki ; Hara, Tomoko ; Hazama, Shunsuke ; Suzuki, Shingo ; Uegaki, Koichi ; Nagappan, Guhan ; Zaitsev, Eugene ; Hirokawa, Takatsugu ; Tatsu, Yoshiro ; Ogura, Akihiko ; Lu, Bai ; Kojima, Masami. / Multiple functions of precursor BDNF to CNS neurons : Negative regulation of neurite growth, spine formation and cell survival. In: Molecular brain. 2009 ; Vol. 2, No. 1.
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abstract = "Background. Proneurotrophins and mature neurotrophins elicit opposite effects via the p75 neurotrophin receptor (p75NTR) and Trk tyrosine kinase receptors, respectively; however the molecular roles of proneurotrophins in the CNS are not fully understood. Results. Based on two rare single nucleotide polymorphisms (SNPs) of the human brain-derived neurotrophic factor (BDNF) gene, we generated R125M-, R127L- and R125M/R127L-BDNF, which have amino acid substitution(s) near the cleavage site between the pro- and mature-domain of BDNF. Western blot analyses demonstrated that these BDNF variants are poorly cleaved and result in the predominant secretion of proBDNF. Using these cleavage-resistant proBDNF (CR-proBDNF) variants, the molecular and cellular roles of proBDNF on the CNS neurons were examined. First, CR-proBDNF showed normal intracellular distribution and secretion in cultured hippocampal neurons, suggesting that inhibition of proBDNF cleavage does not affect intracellular transportation and secretion of BDNF. Second, we purified recombinant CR-proBDNF and tested its biological effects using cultured CNS neurons. Treatment with CR-proBDNF elicited apoptosis of cultured cerebellar granule neurons (CGNs), while treatment with mature BDNF (matBDNF) promoted cell survival. Third, we examined the effects of CR-proBDNF on neuronal morphology using more than 2-week cultures of basal forebrain cholinergic neurons (BFCNs) and hippocampal neurons. Interestingly, in marked contrast to the action of matBDNF, which increased the number of cholinergic fibers and hippocampal dendritic spines, CR-proBDNF dramatically reduced the number of cholinergic fibers and hippocampal dendritic spines, without affecting the survival of these neurons. Conclusion. These results suggest that proBDNF has distinct functions in different populations of CNS neurons and might be responsible for specific physiological cellular processes in the brain.",
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Koshimizu, H, Kiyosue, K, Hara, T, Hazama, S, Suzuki, S, Uegaki, K, Nagappan, G, Zaitsev, E, Hirokawa, T, Tatsu, Y, Ogura, A, Lu, B & Kojima, M 2009, 'Multiple functions of precursor BDNF to CNS neurons: Negative regulation of neurite growth, spine formation and cell survival', Molecular brain, vol. 2, no. 1, 27. https://doi.org/10.1186/1756-6606-2-27

Multiple functions of precursor BDNF to CNS neurons : Negative regulation of neurite growth, spine formation and cell survival. / Koshimizu, Hisatsugu; Kiyosue, Kazuyuki; Hara, Tomoko; Hazama, Shunsuke; Suzuki, Shingo; Uegaki, Koichi; Nagappan, Guhan; Zaitsev, Eugene; Hirokawa, Takatsugu; Tatsu, Yoshiro; Ogura, Akihiko; Lu, Bai; Kojima, Masami.

In: Molecular brain, Vol. 2, No. 1, 27, 24.09.2009.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Multiple functions of precursor BDNF to CNS neurons

T2 - Negative regulation of neurite growth, spine formation and cell survival

AU - Koshimizu, Hisatsugu

AU - Kiyosue, Kazuyuki

AU - Hara, Tomoko

AU - Hazama, Shunsuke

AU - Suzuki, Shingo

AU - Uegaki, Koichi

AU - Nagappan, Guhan

AU - Zaitsev, Eugene

AU - Hirokawa, Takatsugu

AU - Tatsu, Yoshiro

AU - Ogura, Akihiko

AU - Lu, Bai

AU - Kojima, Masami

PY - 2009/9/24

Y1 - 2009/9/24

N2 - Background. Proneurotrophins and mature neurotrophins elicit opposite effects via the p75 neurotrophin receptor (p75NTR) and Trk tyrosine kinase receptors, respectively; however the molecular roles of proneurotrophins in the CNS are not fully understood. Results. Based on two rare single nucleotide polymorphisms (SNPs) of the human brain-derived neurotrophic factor (BDNF) gene, we generated R125M-, R127L- and R125M/R127L-BDNF, which have amino acid substitution(s) near the cleavage site between the pro- and mature-domain of BDNF. Western blot analyses demonstrated that these BDNF variants are poorly cleaved and result in the predominant secretion of proBDNF. Using these cleavage-resistant proBDNF (CR-proBDNF) variants, the molecular and cellular roles of proBDNF on the CNS neurons were examined. First, CR-proBDNF showed normal intracellular distribution and secretion in cultured hippocampal neurons, suggesting that inhibition of proBDNF cleavage does not affect intracellular transportation and secretion of BDNF. Second, we purified recombinant CR-proBDNF and tested its biological effects using cultured CNS neurons. Treatment with CR-proBDNF elicited apoptosis of cultured cerebellar granule neurons (CGNs), while treatment with mature BDNF (matBDNF) promoted cell survival. Third, we examined the effects of CR-proBDNF on neuronal morphology using more than 2-week cultures of basal forebrain cholinergic neurons (BFCNs) and hippocampal neurons. Interestingly, in marked contrast to the action of matBDNF, which increased the number of cholinergic fibers and hippocampal dendritic spines, CR-proBDNF dramatically reduced the number of cholinergic fibers and hippocampal dendritic spines, without affecting the survival of these neurons. Conclusion. These results suggest that proBDNF has distinct functions in different populations of CNS neurons and might be responsible for specific physiological cellular processes in the brain.

AB - Background. Proneurotrophins and mature neurotrophins elicit opposite effects via the p75 neurotrophin receptor (p75NTR) and Trk tyrosine kinase receptors, respectively; however the molecular roles of proneurotrophins in the CNS are not fully understood. Results. Based on two rare single nucleotide polymorphisms (SNPs) of the human brain-derived neurotrophic factor (BDNF) gene, we generated R125M-, R127L- and R125M/R127L-BDNF, which have amino acid substitution(s) near the cleavage site between the pro- and mature-domain of BDNF. Western blot analyses demonstrated that these BDNF variants are poorly cleaved and result in the predominant secretion of proBDNF. Using these cleavage-resistant proBDNF (CR-proBDNF) variants, the molecular and cellular roles of proBDNF on the CNS neurons were examined. First, CR-proBDNF showed normal intracellular distribution and secretion in cultured hippocampal neurons, suggesting that inhibition of proBDNF cleavage does not affect intracellular transportation and secretion of BDNF. Second, we purified recombinant CR-proBDNF and tested its biological effects using cultured CNS neurons. Treatment with CR-proBDNF elicited apoptosis of cultured cerebellar granule neurons (CGNs), while treatment with mature BDNF (matBDNF) promoted cell survival. Third, we examined the effects of CR-proBDNF on neuronal morphology using more than 2-week cultures of basal forebrain cholinergic neurons (BFCNs) and hippocampal neurons. Interestingly, in marked contrast to the action of matBDNF, which increased the number of cholinergic fibers and hippocampal dendritic spines, CR-proBDNF dramatically reduced the number of cholinergic fibers and hippocampal dendritic spines, without affecting the survival of these neurons. Conclusion. These results suggest that proBDNF has distinct functions in different populations of CNS neurons and might be responsible for specific physiological cellular processes in the brain.

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