Journal: The Journal of Neuroscience

Article Title: Differential Distribution of α 2A and α 2C Adrenergic Receptor Immunoreactivity in the Rat Spinal Cord

doi: 10.1523/JNEUROSCI.18-15-05928.1998

Figure Lengend Snippet: Characterization of α2A-AR-IR fibers in rat spinal cord. Single sections of lumbar spinal cord were double-labeled with α2A-AR antisera in combination with various other antisera. The first column(A, E, I,M, Q, U) depicts α2A-AR-IR (red). The second column (B, F, J,N, R, V) depicts immunoreactivity obtained with other antisera used in double-labeling (green). The third column(C, G, K,O, S, W) depicts digitally merged images of the first two columns. In merged images, the appearance of yellow may indicate colocalization, although higher magnification is required to distinguish colocalization from close apposition. The fourth column(D, H, L,P, T, X) depicts merged images obtained at higher magnification. Row 1(A–D) demonstrates that α2A-AR -IR and α2C-AR-IR are rarely colocalized. Row 2(E–H) demonstrates that α2A-AR-IR and SP-IR are highly colocalized. Row 3 (I–L) demonstrates that α2A-AR-IR and CGRP-IR fibers are highly, but not exclusively, colocalized. Row 4(M–P) demonstrates that α2A-AR-IR and ENK-IR are rarely colocalized. Row 5 (Q–T) demonstrates that α2A-AR-IR and DβH-IR are closely apposed, but not colocalized. Similarly, row 6(U–X) demonstrates that instances of colocalization between α2A-AR-IR and TH-IR could not be detected. SP, Substance P; CGRP, calcitonin gene-related peptide; ENK, leuenkephalin; DβH, dopamine β-hydroxylase; TH, tyrosine hydroxylase.

Article Snippet: Primary antisera were used at the following dilutions: rabbit anti-α 2A -AR, 1:1000; rabbit anti-α 2C -AR, 1:1000; guinea pig anti-α 2C -AR, 1:500; mouse anti-calcitonin gene-related peptide (CGRP), 1:2000 (Research Biochemicals, Natick, MA); mouse anti-dopamine-β-hydroxylase, 1:500 (Chemicon, Temecula, CA); rat anti-substance P (SP), 1:1000 (Research Biochemicals); rabbit anti-neurokinin-1 receptor (NK-1R) 1:500 ( Vigna et al., 1994 ; a gift from Dr. P. Mantyh, University of Minnesota, Minneapolis, MN); rabbit anti-somatostatin, 1:1000 ( Seybold and Elde, 1980 ); mouse anti-leuenkephalin, 1:500 (Sera-Lab, Sussex, UK); mouse anti-tyrosine hydroxylase, 1:500 (IncStar, Stillwater, MN); guinea pig anti-preprodynorphin, 1:500 ( Arvidsson et al., 1995b ); and mouse anti-GFAP, 1:500 (Sigma).

Techniques: Labeling

Journal: The Journal of Neuroscience

Article Title: Differential Distribution of α 2A and α 2C Adrenergic Receptor Immunoreactivity in the Rat Spinal Cord

doi: 10.1523/JNEUROSCI.18-15-05928.1998

Figure Lengend Snippet: Characterization of α2C-AR-IR fibers in rat spinal cord. Single sections of lumbar spinal cord were double-labeled with α2C-AR antisera in combination with various other antisera. The first column(A, E, I,M, Q, U,Y) depicts α2C-AR-IR (red). The second column(B, F, J,N, R, V, Z) depicts immunoreactivity obtained with other antisera used in double-labeling (green). The third column (C, G, K,O, S, W,AA) depicts digitally merged images of the first two columns. In merged images, the appearance of yellow may indicate colocalization, although higher magnification is required to distinguish colocalization from close apposition. The fourth column (D, H, L,P, T, X,BB) depicts merged images obtained at higher magnification. Row 1(A–D) demonstrates that α2C-AR-IR and SP-IR are rarely colocalized. Row 2 (E–H) demonstrates that α2C-AR-IR and CGRP-IR fibers are moderately colocalized.Row 3 (I–L) demonstrates that α2C-AR-IR and somatostatin-IR fibers are moderately colocalized. Row 4 (M–P) demonstrates that α2C-AR-IR and ENK-IR are moderately colocalized. Row 5(Q–T) demonstrates that α2C-AR-IR and NK-1R are not colocalized. Row 6 (U–X) demonstrates that α2C-AR-IR and GFAP-IR are very rarely colocalized.Row 7 (Y–BB) demonstrates that α2C-AR-IR and DβH-IR are not colocalized. A similar lack of colocalization was observed between α2C-AR-IR and TH-IR (data not shown). SP, Substance P; CGRP, calcitonin gene-related peptide; SOM, somatostatin; ENK, leuenkephalin; NK-1R, NK-1 receptor; GFAP, glial fibrillary acidic protein; DβH, dopamine β-hydroxylase.

Article Snippet: Primary antisera were used at the following dilutions: rabbit anti-α 2A -AR, 1:1000; rabbit anti-α 2C -AR, 1:1000; guinea pig anti-α 2C -AR, 1:500; mouse anti-calcitonin gene-related peptide (CGRP), 1:2000 (Research Biochemicals, Natick, MA); mouse anti-dopamine-β-hydroxylase, 1:500 (Chemicon, Temecula, CA); rat anti-substance P (SP), 1:1000 (Research Biochemicals); rabbit anti-neurokinin-1 receptor (NK-1R) 1:500 ( Vigna et al., 1994 ; a gift from Dr. P. Mantyh, University of Minnesota, Minneapolis, MN); rabbit anti-somatostatin, 1:1000 ( Seybold and Elde, 1980 ); mouse anti-leuenkephalin, 1:500 (Sera-Lab, Sussex, UK); mouse anti-tyrosine hydroxylase, 1:500 (IncStar, Stillwater, MN); guinea pig anti-preprodynorphin, 1:500 ( Arvidsson et al., 1995b ); and mouse anti-GFAP, 1:500 (Sigma).

Techniques: Labeling