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Table 1 Astroglial and microglial NOS2 immunoreactivity in LPS-activated mouse/rat astroglial-enriched/mixed glial cultures. This table summarizes all the studies that, to my knowledge, have addressed the question of what cell types express NOS2 in LPS-activated rodent astroglial-enriched or mixed glial cultures. Studies using activating stimuli other than LPS or studies on human glial cultures are not included in this table. Reports describing the presence of NADPH diaphorase in LPS-treated astroglial-enriched cultures (e.g [77]) are also not included because NOS2 is only one of many brain enzymes that exhibit NADPH diaphorase activity [78].

From: Microglial cells in astroglial cultures: a cautionary note

Ref.

Species-Age-Region

[LPS] μg/ml

Co-activators

Time post-LPS

Presence in astrocytes*

Presence in microglia (marker)

Notes

Double labelling method**

[29] Fig 4

Mouse-Neonatal-Whole Brain

1

-

-

Yes (?)

Yes (Mac1)

"The number of GFAP/NOS2 positive cells was low". NOS2(+) cells have microglial morphology and not a clear GFAP staining

DAB-Ni and DAB

[68] Fig 2

Rat-Neonatal-Cortex

100

-

24 h

No

-

NOS2(+) cells have a microglial appearance but no double labelling is done

IF

[69] Fig 1

Rat-Neonatal-Cortex

1

-

24 h

No

Yes (Isolectin-B4)

 

IF

[70] Fig 1

Rat-Neonatal-Forebrain

1

-

12 h, 24 h, 48 h

No

-

NOS2(+) cells have a microglial appearance but no double labelling NOS2/microglia is done

DAB and AP

[71] Fig 3

Rat-Neonatal-Cortex

2

IFNγ (100 U/ml)

18 h

No

Yes (OX-42)

 

IF

[72] Fig 2

Rat-Neonatal-Cortex

2

IFNγ (100 U/ml)

18 h

No

Yes (OX-42)

 

IF

[73] Fig 3

Rat-Neonatal-Cortex

2

IFNγ (100 U/ml)

18 h

-

Yes (OX-42)

"All NOS2(+) cells were OX-42(+)"

IF

[74] Fig 2

Rat-Neonatal-Cortex

2

IFNγ (100 U/ml)

18 h

No

-

 

IF

[39] Figs 2,3,5

Rat-Neonatal-Cortex

10

-

48 h

No

Yes (OX-42)

 

IF

[30] Fig 3

Rat-Neonatal-Neopallium

0,025

IFNγ (100 U/ml)

24 h

Yes (?)

-

Some double GFAP-NOS2(+) cells do not look like astrocytes

IF

[75] Fig 5

Rat-Neonatal-Cortex

1

IFNg (100 U/ml)

48 h

No

Yes (Isolectin-B4)

 

IF

[28] Fig 1

Rat-Neonatal-Spinal cord

1

-

24 h

Yes

-

Double GFAP-NOS2(+) cells. Double NOS2-microglial staining not done because "these cultures had no OX42 positive cells"

IF

[76] Fig 3

Rat-Neonatal-Cortex

0,1

-

24 h

No

Yes (OX-42)

"All NOS2(+) cells were OX-42(+)"

IF

[27] Fig 2

Rat-Neonatal-Cortex

1

IFNγ (10 ng/ml)

14 h

Yes

Yes (Isolectin-B4)

"NOS2 staining is much stronger in microglia than in astrocytes"

IF

[24] Fig 6

Mouse-Neonatal-Cortex

1

CGS21680 (100 nM)

48 h

No

Yes (Tomato lectin)

 

IF

[26] Fig 6

Mouse-Neonatal-Cortex

2

IFNγ (3 ng/ml)

24 h

Yes

-

Double GFAP-NOS2(+) cells

IF

[25] Fig 8

Mouse-Neonatal-Cortex

2

IFNγ (3 ng/ml) and TGFβ1 (3 ng/ml)

14 h

Yes

-

Double GFAP-NOS2(+) cells. The number of NOS2(+) astrocytes is very low after LPS but high after TGFβ1 + LPS + IFNγ treatment

IF

  1. * GFAP was the astroglial marker used in all the studies
  2. ** DAB: diaminobenzidine, DAB-Ni: DAB-Nickel; AP: Alkaline phosphatase; IF: immunofluorescence