Neurochemical Research

, Volume 35, Issue 2, pp 254–261 | Cite as

Postnatal Expression of GAD67

  • Christopher P. Turner
  • Emily Ware
  • Robert Stowe
  • Danielle DeBenedetto
  • Caroline Walburg
  • Andrew Lee
  • John Swanson
  • Alexandra Lambert
  • Melissa Lyle
  • Priyanka Desai
  • Chun Liu
Original Paper

Abstract

N-methyl-d-aspartate receptor blockade promotes apoptosis at postnatal day 7 (P7) and is linked to loss of glutamic acid decarboxylase 67 (GAD67) expression in older animals. To more fully appreciate this relationship we must first understand how GAD67 is regulated postnatally. Thus, the brains of P7, P14 and P21 rats were examined for expression of GAD67 protein and we found that levels of this GABAergic marker increased steadily with age, such that by P21 there was as much as a 6-fold increase compared to P7 animals and a 1.5- to 2-fold increase compared to P14 animals, depending on the region sampled. At P7, GAD67 was almost exclusively detected in puncta, with very few cell bodies displaying this marker. In contrast, at P14 and especially P21, both puncta and cell bodies were robustly labeled. Our data indicate that adult-like expression of GAD67 emerges quite late in the postnatal period.

Keywords

Rat Postnatal Somatosensory Cingulate Caudate-putamen GABA 

Abbreviations

P7

Postnatal day 7

P14

Postnatal day 14

P21

Postnatal day 21

GAD67

Glutamic acid decarboxylase 67

NMDAR

N-methyl-d-aspartate receptor

AC3

Activated caspase-3

CaBP

Calcium binding proteins

PV

Parvalbumin

Cg

Cingulate cortex

SSC

Somatosensory cortex

CP

Caudate putamen

GP

Globus pallidus

TRN

Thalamic reticular nucleus

ir

Immunoreactivity

Supplementary material

11064_2009_49_MOESM1_ESM.tif (2.2 mb)
Supplementary Fig. 1Developmental changes in GAD67 expression: posterior forebrain. GAD67 expression in posterior forebrain at: (a) P7 (b) P14 and (c) P21. RS—retrosplenial cortex; SSCPOST—posterior somatosensory cortex; CC—corpus callosum; Hip—hippocampus; TRN—thalamic reticular nucleus (extent indicated by arrows); IC—internal capsule; GP—globus pallidus; Hyp—hypothalamus; scale in C is 1 mm. In the hippocampal formation, the CA3 division as well as the dentate gyrus showed heavy GAD67-ir at P7 but at later ages far more of this structure was labeled. In some structures, such as the GP or the TRN, robust staining was observed even at P7. (TIFF 2207 kb)
11064_2009_49_MOESM2_ESM.tif (2.4 mb)
Supplementary Fig. 2Punctate and somatic distribution of GAD67: cingulate cortex. GAD67 expression in cingulate cortex at: (a–c) P7 (d–f) P14 and (g–i) P21. Roman numerals indicate the layer each panel represents. Scale in I indicates 25 µm. Please note that at P7 there is little evidence of somatic GAD67. However, light punctate staining was observed (arrowheads). At P14, somatic staining was now clearly visible (arrows) and punctate immunoreactivity was greater than that found at P7 (arrowheads). At P21, somatic and punctate staining for GAD67 was similar to that found at P14. It should be noted that at P21 it was often difficult to distinguish cell bodies from the intense punctate staining observed in many of the layers (TIFF 2439 kb)
11064_2009_49_MOESM3_ESM.tif (2.4 mb)
Supplementary Fig. 3Punctate and somatic distribution of GAD67: somatosensory cortex. GAD67 expression in cingulate cortex at: (a–c) P7, (d–f) P14 and (g–i) P21. Roman numerals indicate the layer each panel represents. Scale in I indicates 25 µm. Please note that at P7 there is little evidence of somatic GAD67. However, light punctate staining was observed (arrowheads). At P14, somatic staining was now clearly visible (arrows) and punctate immunoreactivity was greater than that found at P7 (arrowheads). At P21, somatic and punctate staining for GAD67 was similar to that found at P14. In some areas somatic details were often difficult to distinguish from GAD67-rich puncta (TIFF 2439 kb)
11064_2009_49_MOESM4_ESM.tif (2.4 mb)
Supplementary Fig. 4Punctate and somatic distribution of GAD67 in the caudate-putamen. GAD67 expression in the caudate-putamen at: (ac) P7, (df) P14 and (gi) P21. Images were taken from the lateral (Lat; a, d, g), the ventral (Vent; b, e, h) and the dorsomedial areas (DM; c, f, i). Scale in i indicates 25 µm. At P7, although somatic staining was evident much of the GAD67-ir was localized to puncta (arrowheads). At P14, somatic staining was now clearly visible (arrows) and punctate immunoreactivity greater (arrowheads), especially in the DM quadrant. At P21, somatic and punctate GAD67 distribution was similar to that found at P14. As with other brain regions, cellular details were sometimes obscured by the intense immunoreactivity associated with GAD67-positive puncta (TIFF 2413 kb)

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Christopher P. Turner
    • 1
  • Emily Ware
    • 1
  • Robert Stowe
    • 1
  • Danielle DeBenedetto
    • 1
  • Caroline Walburg
    • 1
  • Andrew Lee
    • 1
  • John Swanson
    • 1
  • Alexandra Lambert
    • 1
  • Melissa Lyle
    • 1
  • Priyanka Desai
    • 1
  • Chun Liu
    • 1
  1. 1.Neurobiology and AnatomyWake Forest University School of MedicineWinston-SalemUSA

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