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Rab proteins constitute the largest family of monomeric GTPases, of which 70 members are present in humans. Rab proteins and their effectors regulate consecutive stages of vesicle transport such as vesicle transport, docking, and fusion to the correct recipient membrane.
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Rab GTPases act in a regulated cascade during membrane fusion, helping the lipid bilayers mix. The Rab family of proteins are active when bound to GTP, and inactive when bound to GDP. Hence, they act as guanine nucleotide-dependent molecular switches. Rab-GTP recognizes and binds to long or short-range tethering proteins to capture the target vesicle. These tethers coordinate with SNAREs on the vesicle and the target membrane to assemble the trans SNARE complex that locks the mixing bilayers.
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Complex microtubule structures are present in resting cells and in dividing cells. In resting cells, they are responsible for maintaining the cellular architecture, tracks for intracellular transport, positioning of organelles, assembly of cilia and flagella. They mediate the bipolar spindle assembly for chromosomal segregation and positioning of the cell division plate in dividing cells. The formation of microtubule complex structures depends on the cell type, cell stage, and cell function.
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The primary cilium, made up of microtubules, acts as antennae on the cell surfaces for relaying external stimuli into the cells. These fine hair-like structures are present, generally one per cell. These are non-motile cilia in a 9+0 microtubules arrangement, where the central pair of microtubules are absent. The primary cilia arise from the basal body embedded in the cell membrane. Intraflagellar transport (IFT) carries requisite proteins from the cytoplasm to the cilium because the primary...
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Standardized Method to Detect Tunneling Nanotubes in Human Skin Cells for Tissue Engineering Applications
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Rab8a/Rab11a regulate intercellular communications between neural cells via tunneling nanotubes.

Hui Zhu1,2,3, Chengbin Xue1,2, Xi Xu4

  • 1State Key Laboratory of Pharmaceutical Biotechnology and MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing Biomedical Research Institute, Nanjing University, Nanjing, China.

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Tunneling nanotubes (TNTs) facilitate cell communication and transfer of materials. These structures, regulated by Rab8a/Rab11a, are crucial for peripheral nerve regeneration and Schwann cell survival.

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Area of Science:

  • Cell Biology
  • Neuroscience
  • Regenerative Medicine

Background:

  • Tunneling nanotubes (TNTs) are actin-based membrane structures facilitating intercellular communication, including organelle transfer.
  • Peripheral nerve injury necessitates complex cell communication for effective regeneration.
  • The presence and role of TNTs in the peripheral nervous system (PNS) remain largely unexplored.

Purpose of the Study:

  • To investigate the existence and function of TNTs between Schwann cells (SCs) in the PNS.
  • To determine the impact of TNTs on neural regeneration following injury.
  • To identify molecular regulators of TNT formation in SCs.

Main Methods:

  • Cultured Schwann cells (SCs) were used to study TNT formation and intercellular transfer.
  • Serum depletion was employed to induce TNT-like structures in SCs.
  • Rab8a and Rab11a were downregulated using genetic manipulation (siRNA/shRNA) to assess their role in TNT formation and function.
  • Axonal outgrowth assays with dorsal root ganglion (DRG) neurons were performed.
  • TNT-like structures were examined in sciatic nerve stumps from transected rats.

Main Results:

  • Functional TNTs were observed between cultured SCs, mediating the transfer of proteins, mitochondria, and RNA.
  • TNT-like structures increased in SCs under serum depletion and in injured rat sciatic nerves.
  • Downregulation of Rab8a or Rab11a inhibited TNT formation, vesicle transfer, SC migration, and increased SC apoptosis.
  • Knockdown of Rab8a or Rab11a in SCs suppressed axonal outgrowth from co-cultured DRG neurons.

Conclusions:

  • Rab8a and Rab11a are implicated in the formation of TNTs within the PNS.
  • TNTs play a significant role in peripheral nerve regeneration by regulating intercellular communication between neural and non-neural cells.
  • Targeting TNTs or their regulators may offer therapeutic strategies for enhancing nerve repair.