PMID- 37407448
OWN - NLM
STAT- MEDLINE
DCOM- 20230804
LR  - 20240102
IS  - 1347-7439 (Electronic)
IS  - 0916-7250 (Print)
IS  - 0916-7250 (Linking)
VI  - 85
IP  - 8
DP  - 2023 Aug 1
TI  - Oxygen-glucose deprivation-induced glial cell reactivity in the rat primary 
      neuron-glia co-culture.
PG  - 799-808
LID - 10.1292/jvms.23-0175 [doi]
AB  - It has been demonstrated that in vivo brain ischemia induces activation and 
      proliferation of astrocytes and microglia. However, the mechanism underlying the 
      ischemia-induced activation and proliferation of these cells remains to be 
      unclear. Oxygen-glucose deprivation (OGD), an in vitro ischemia mimic, has been 
      extensively used to analyze the hypoxia response of various cell types. This 
      study examined the OGD-induced changes in the expression level of astrocytes and 
      microglia marker proteins and immunoreactivity for Ki-67, a marker protein for 
      cell proliferation, using rat primary hippocampal neuron-glia co-culture (NGC) 
      cells. Furthermore, OGD-induced changes in the expression of M1/M2 microglia 
      phenotype-related genes were also examined. MTT assay indicated that 120 min of 
      OGD decreased cell viability, and immunocytochemistry indicated that 120 min of 
      OGD abolished most microtubule-associated protein 2 (MAP2)-immunopositive 
      neurons. In contrast, glial fibrillary acidic protein (GFAP)-immunopositive 
      astrocytes and ionized calcium-binding adapter protein-1 (Iba-1)-immunopositive 
      microglia, and 2',3'-cyclic nucleotide-3'-phosphodiesterase 
      (CNPase)-immunopositive oligodendrocytes survived OGD. Western blot assays and 
      double-immunofluorescent staining indicated that OGD increased the GFAP 
      expression level and the Ki-67-immunopositive/GFAP-immunopositive cells' ratio. 
      Real-time PCR analysis showed that OGD altered M1 microglia phenotype-related 
      genes. Specifically, OGD decreased the expression level of CD32 and 
      interleukin-1beta (IL-1beta) genes and increased that of the inducible nitric oxide 
      synthase (iNOS) gene. Therefore, applying OGD to NGC cells could serve as a 
      useful in vitro tool to elucidate the molecular mechanisms underlying brain 
      ischemia-induced changes in GFAP expression, astrocyte proliferation, and M1 
      microglia phenotype-related gene expression.
FAU - Inoue, Maiko
AU  - Inoue M
AD  - Laboratory of Veterinary Anatomy, Graduate School of Veterinary Science, Osaka 
      Metropolitan University, Osaka, Japan.
FAU - Tanida, Takashi
AU  - Tanida T
AD  - Laboratory of Veterinary Anatomy, Graduate School of Veterinary Science, Osaka 
      Metropolitan University, Osaka, Japan.
FAU - Kondo, Tomohiro
AU  - Kondo T
AD  - Laboratory of Animal Science, Graduate School of Veterinary Science, Osaka 
      Metropolitan University, Osaka, Japan.
FAU - Takenaka, Shigeo
AU  - Takenaka S
AD  - Department of Nutrition, Graduate School of Human Life and Ecology, Osaka 
      Metropolitan University, Osaka, Japan.
FAU - Nakajima, Takayuki
AU  - Nakajima T
AD  - Laboratory of Veterinary Anatomy, Graduate School of Veterinary Science, Osaka 
      Metropolitan University, Osaka, Japan.
LA  - eng
PT  - Journal Article
DEP - 20230706
PL  - Japan
TA  - J Vet Med Sci
JT  - The Journal of veterinary medical science
JID - 9105360
RN  - S88TT14065 (Oxygen)
RN  - IY9XDZ35W2 (Glucose)
RN  - 0 (Ki-67 Antigen)
SB  - IM
MH  - Rats
MH  - Animals
MH  - Coculture Techniques/veterinary
MH  - Oxygen/metabolism
MH  - Glucose/metabolism
MH  - Ki-67 Antigen/metabolism
MH  - Neurons/metabolism
MH  - Astrocytes
MH  - *Brain Ischemia/metabolism/veterinary
MH  - Microglia
PMC - PMC10466061
OTO - NOTNLM
OT  - astrocytes
OT  - microglia
OT  - neuron-glia co-culture
OT  - oxygen-glucose deprivation
OT  - rat
COIS- The authors declare no conflict of interest associated with this manuscript.
EDAT- 2023/07/06 01:08
MHDA- 2023/08/04 06:43
PMCR- 2023/08/01
CRDT- 2023/07/05 22:13
PHST- 2023/08/04 06:43 [medline]
PHST- 2023/07/06 01:08 [pubmed]
PHST- 2023/07/05 22:13 [entrez]
PHST- 2023/08/01 00:00 [pmc-release]
AID - 23-0175 [pii]
AID - 10.1292/jvms.23-0175 [doi]
PST - ppublish
SO  - J Vet Med Sci. 2023 Aug 1;85(8):799-808. doi: 10.1292/jvms.23-0175. Epub 2023 Jul 
      6.