MAP-2-A Versatile Neuron Marker

Neuromics is a leader in providing Neuron-Glial Markers for Neuroscientists.

We are constantly on the search for publications that reference use of these markers in unique applications. In this posting I would like to share a publication where researchers used on of our MAP-2 antibodies to stain medial superior olive (MSO) neurons. Baumann Veronika, Lehnert Simon, Leibold Christian, Koch Ursula. Tonotopic Organization of the Hyperpolarization-activated Current (Ih) in the Mammalian Medial Superior Olive. Front. Neural Circuits 7:117. doi: 10.3389/fncir.2013.00117.
 ...Following recording, slices were fixed in 4% paraformaldehyde for 30 min. After extensive washing in phosphate-buffered saline (PBS) slices were exposed to blocking buffer (0.5% trition X-100/0.1% saponin/1% BSA in PBS) followed by incubation with the primary antibody (chicken anti-microtubule-associated protein 2, MAP2, 1:1000, Neuromics) in blocking buffer. Slices were then rinsed in washing buffer (0.5% Trition X-100/0.1% saponin in PBS) and immunoreactivity was visualized by incubating the slices with the Cy3-conjugated secondary antibody raised in donkey (1:300; Dianova). Finally, slices were washed and mounted on slides with vectashield mounting reagent (Vector Laboratories, USA)...

Here the MAP-2 antibody is used to help identify the dorsal, medial and ventral portion of the MSO of p18 and p22 gerbils.

Figure . Ih varies systematically along the dorsoventral axis. (A) A brain slice containing the MSO with Alexa-488-filled neurons (green) verifies the distribution of the patched neurons along the dorsoventral axis (red: MAP-2). (B) Pharmacologically isolated Ih current traces were elicited by depolarizing and hyperpolarizing voltage steps from −60.5 mV to potentials between −40.5 mV and −120.5 mV for 1 s in 5 mV step increment and then to −100.5 mV for 0.5 s to elicit the tail current to determine the voltage dependence of Ih activation. Current traces are representative for the dorsal, the intermediate and the ventral part of the MSO. (C) I-V relationships of steady-state (red arrow in B) Ih density for ventral (n = 15), intermediate (n = 12) and dorsal (n = 18) neurons emphasize that Ih density amplitudes are smallest in dorsal neurons and largest in ventral neurons (C1). Ih density amplitudes for a voltage step to −110.5 mV (C2). (D) Weighted activation time constants at −110.5 mV (D1). The weighted activation time constants are voltage dependent and largest in the dorsal part of the MSO (D2). (E) The voltage-dependence of Ih activation was measured from the tail current 20 ms after the end of the voltage steps (red arrow) (E1). Values were fitted with a Boltzmann function to obtain the half-maximal activation voltage. In dorsal neurons the Ih activation curve is shifted to more negative voltages (E2). Half-maximal activation voltage was measured in each experiment and averaged (E3). Black symbols: dorsal neurons; gray symbols: intermediate neurons; white symbols: ventral neurons. **P < 0.01, ***P < 0.001, single-factor ANOVA test followed by a Scheffe's post-hoc test.

I will continue to post interesting applications using our Neuron-Glial Markers.

Markers for Medial Superior Olivary Neurons

This is an excellent reference for researchers looking for immunohistochemistry images of slice preparations of the Neurons in the medial superior olive (MSO). It also references use of our widely used and frequently published MAP2 (Microtubule associated protein 2).

Kiri Couchman, Benedikt Grothe and Felix Felmy. Medial Superior Olivary Neurons Receive Surprisingly Few Excitatory and Inhibitory Inputs with Balanced Strength and Short-Term Dynamics. The Journal of Neuroscience, December 15, 2010, 30(50):17111-17121; doi:10.1523/JNEUROSCI.1760-10.2010.

Summary: Neurons in the medial superior olive (MSO) process microsecond interaural time differences, the major cue for localizing low-frequency sounds, by comparing the relative arrival time of binaural, glutamatergic excitatory inputs. This coincidence detection mechanism is additionally shaped by highly specialized glycinergic inhibition. Traditionally, it is assumed that the binaural inputs are conveyed by many independent fibers, but such an anatomical arrangement may decrease temporal precision. Short-term depression on the other hand might enhance temporal fidelity during ongoing activity. For the first time we show that binaural coincidence detection in MSO neurons may require surprisingly few but strong inputs, challenging long-held assumptions about mammalian coincidence detection. This study exclusively uses adult gerbils for in vitro electrophysiology, single-cell electroporation and immunohistochemistry to characterize the size and short-term plasticity of inputs to the MSO. We find that the excitatory and inhibitory inputs to the MSO are well balanced both in strength and short-term dynamics, redefining this fastest of all mammalian coincidence detector circuits.

Related Reagents:
Neuronal-Glial Markers-Astrocytes, Glia,
Microglia, Olidogodendrocytes, Progenitors and Schwann Cell Markers
Stem Cell Research Antibodies
Stem Cell Research Reagents
Primary Neurons and Astrocytes-Primary

human, rat and mouse neurons and astrocytes.

Potent Neuron-Glial Markers

We are recognized for having top shelf Neuron/Glial Marker Antibodies. We have an extensive catalog and have customer referencing use of these in a variety of applications, species and cell types.

Cell types include neural progenitors, neurons, glia, astrocytes, schwann cells and more. We are pleased to provide present a new publication referencing use of our MAP2 (Microtubule assoc. protein 2) Antibody for immunostaining of E17 primary mouse astrocytes.

Shelley Jacobs and Laurie C. Doering. Astrocytes Prevent Abnormal Neuronal Development in the Fragile X Mouse. J. Neurosci., Mar 2010; 30: 4508 - 4514 ; doi:10.1523/JNEUROSCI.5027-09.2010.

After 7 d in vitro (DIV), the cells were fixed with ice-cold (–20°C) methanol and processed for immunocytochemistry. After the appropriate serum block, the cells were incubated with primary antibodies overnight at 4°C. Secondary antibodies were applied for 3 h at room temperature. The following antibody, diluted in 1% BSA, was used: chicken microtubule-associated protein 2 (MAP2) (1:20,000; Neuromics) and anti-chicken FITC (1:100; Jackson ImmunoResearch Laboratories). Coverslips were mounted with Vectashield fluorescent mounting medium with 4`,6-diamidino-2-phenylindole (DAPI).

Images: Effects of astrocytes on the growth of hippocampal neurons in coculture at 7 DIV. E17 primary hippocampal neurons were cocultured with P0–P1 primary cortical astrocytes for 7 DIV in each of four coculture conditions. a, Immunofluorescent images of neurons in each of the four culture combinations. Neurons are stained with an antibody directed against the neuronal dendritic marker, MAP2. Scale bar, 100 µm. b, Quantification of percentage of surviving neurons at 7 DIV in each of the four culture conditions. Data shown are mean values ± SEM from two or three independent experiments (10–15 regions of 1.5 mm2 from 2 coverslips per experiment). Significant differences revealed by post hoc Tukey's tests are indicated (p less than 0.001).

Related Reagents

• Stem Cell Research Antibodies


• Stem Cell Research Reagents


• Primary Neurons and Astrocytes-Primary
  human, rat and mouse neurons and astrocytes