Related Experiment Video

Updated: Aug 9, 2026

Isolation of CA1 Nuclear Enriched Fractions from Hippocampal Slices to Study Activity-dependent Nuclear Import of Synapto-nuclear Messenger Proteins
10:03

Isolation of CA1 Nuclear Enriched Fractions from Hippocampal Slices to Study Activity-dependent Nuclear Import of Synapto-nuclear Messenger Proteins

Published on: August 10, 2014

A phosphorylation epitope on MAP 1B that is transiently expressed in growing axons in the developing rat nervous

P R Gordon-Weeks1, S G Mansfield, C Alberto

  • 1Developmental Biology Research Centre, King's College London, UK.

The European Journal of Neuroscience
|October 1, 1993
PubMed

Insights

A new monoclonal antibody (150) identifies a specific phosphorylated epitope on microtubule-associated protein 1B (MAP 1B). This epitope is transiently found in developing rat axons, suggesting a role in axonogenesis.

Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Microtubule-associated protein 1B (MAP 1B) is crucial for microtubule dynamics.
  • Phosphorylation of MAP 1B regulates its function.
  • Understanding MAP 1B phosphorylation in vivo is essential for comprehending neural development.

Purpose of the Study:

  • To isolate and characterize a monoclonal antibody targeting a specific phosphorylation epitope on MAP 1B.
  • To investigate the in vivo expression and localization of this epitope in the developing rat nervous system.
  • To determine the potential role of this phosphorylated epitope in axonogenesis.

Main Methods:

  • Isolation of a monoclonal antibody (mAb 150) against a phosphorylated epitope on MAP 1B.
  • Immunoblot and immunoprecipitation analyses to confirm antibody specificity and epitope presence on MAP 1B.
  • Alkaline phosphatase treatment to verify the phosphorylated nature of the epitope.
  • Immunohistochemistry on developing rat central nervous system sections to determine epitope localization and temporal expression.

Main Results:

  • Monoclonal antibody 150 specifically recognizes a phosphorylated epitope on the 325 kDa MAP 1B.
  • The epitope is present on MAP 1B that copolymerizes with microtubules.
  • Immunohistochemistry revealed transient expression of the epitope in growing axons, radial glial fibers, and some cell nuclei, but not in dendrites.
  • Alkaline phosphatase treatment abolished all immunostaining, confirming the epitope's phosphorylated state.

Conclusions:

  • A novel phosphorylation epitope on MAP 1B, recognized by mAb 150, has been identified in vivo.
  • The transient expression pattern of this epitope in developing axons suggests a critical role in axonogenesis.
  • This finding provides new insights into the regulation of MAP 1B during neural development.

More Related Videos

Rapid Genotyping of Animals Followed by Establishing Primary Cultures of Brain Neurons09:51

Rapid Genotyping of Animals Followed by Establishing Primary Cultures of Brain Neurons

Published on: January 29, 2015

Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons10:13

Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons

Published on: February 10, 2016

Related Concept Videos

Enzyme-linked Receptors01:00

Enzyme-linked Receptors

Enzyme-linked receptors are proteins that act as both receptor and enzyme, activating multiple intracellular signals. This is a large group of receptors that include the receptor tyrosine kinase (RTK) family. Many growth factors and hormones bind to and activate the RTKs.
Neurotrophin (NT) receptors are a family of RTKs, including trkA, trkB, and trkC (tropomyosin-related kinase) receptors. TrkA is specific for nerve growth factor (NGF), neurotrophin-6, and neurotrophin-7. TrkB binds...
Microtubule Associated Proteins (MAPs)01:42

Microtubule Associated Proteins (MAPs)

Microtubule function and architecture are regulated by an array of specialized proteins called microtubule-associated proteins or MAPs. These proteins are widespread across different organisms and have conserved protein motifs, like the multi-TOG domain for tubulin binding found in the CLASP family of MAPs. Some MAPs are lineage-specific based on their conserved domains. Their functions depend upon the cytoskeletal architecture and cell type they are located within. In-plant cells, a specific...
MAPK Signaling Cascades01:07

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
Calmodulin-dependent Signaling01:16

Calmodulin-dependent Signaling

Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...