Skip to main content
. Author manuscript; available in PMC: 2017 Jun 15.
Published in final edited form as: J Comp Neurol. 2016 Jan 4;524(9):1892–1919. doi: 10.1002/cne.23946

Figure 1. Specificity of calbindin, parvalbumin and VGLUT1 antibodies tested in knockout tissues.

Figure 1

All images are in epifluorescence captured at low magnification with a digital camera. with All images are wide-field epifluorescence captured at low magnification with a digital camera. A-B, Parvalbumin and calbindin-immunoreactivity in the cerebellum of wild-type (A1 and B1), parvalbumin knockout (A2) and calbindin knockout (B2) adult mice. Parvalbumin (A1) and calbindin (B1) immunoreactivities are revealed in Purkinje cells of adult wild-type mice and abolished in the cerebellum of their respective knockout animals (A2, B2)(dashed line indicate the Purkinje cell layer). C-D, Parvalbumin and calbindin-immunoreactivities in the spinal cord of adult wild-type (C1 and D1), parvalbumin knockout (C2) and calbindin knockout (D2) mice. Parvalbumin-IR afferent axons are visible in the dorsal columns (DCs) of the spinal cord of wild-type mice (C1, outlined area), but not in parvalbumin knockout mice (C2). Calbindin antibodies reveal in wild-type spinal cords a large number of immunoreactive cells in lamina II (LII, D1) and in the Renshaw cell area (ventral dashed outline, D1). No immunoreactive neurons are observed in spinal cord sections from knockout animals (D2). E, VGLUT1-immunoreactivity in the spinal cord of wild-type (E1), VGLUT1 heterozygote (E2) and VGLUT1 knockout (E3) mice. A normal pattern of VGLUT1-IR boutons is present in spinal cords from wild type (E1) and heterozygotes (E2). There are no VGLUT1-IR boutons detected in VGLUT1 knockout mice (E3). Some blood vessel immunolabeling remains in knockout mice indicating it is due to a cross-reaction of the primary antibody (it is not revealed by secondary antibodies applied to the section in the absence of primary antibodies). This relatively weaker immunoreaction of blood vessels did not interfere with the identification of VGLUT1-IR synaptic puncta on spinal neurons and was variable from animal to animal. It is also a particular feature of the guinea-pig antibody against VGLUT1 we used since other anti-VGLUT1 antibodies raised in different species (rabbit) did not show blood vessel labeling. We used guinea pig antibodies to better combine with other rabbit-raised primary antibodies for dual and triple immunfluorescence. Scale bars: 200 μm in A1, B1, C1, D1 and E1.