title: Activation of the neurokinin 3 receptor promotes filopodia growth and sprouting in rat embryonic hypothalamic cells.
abstract: Members of the tachykinin family have trophic effects on developing neurons. The tachykinin neurokinin 3 receptor (NK3R) appears early in embryonic development; during the peak birthdates of hypothalamic neurons, but its involvement in neural development has not been examined. To address its possible role, immortalized embryonic hypothalamic neurons (CLU209) were treated with CellMask, a plasma membrane stain, or the membranes were imaged in CLU209 cells that were transfected with a pEGFP-NK3R expression vector. Nontransfected cells and transfected cells were then treated with senktide, a NK3R agonist, or Dulbecco's Modified Eagle's Medium (DMEM) and time-lapse confocal images were captured for the following 30 min. Compared to DMEM, senktide treatment led to filopodia initiation from the soma of both nontransfected and transfected CLU209 cells. These filopodia had diameters and lengths of approximately 200 nm and 3 µm, respectively. Pretreatment with an IP3 receptor blocker, 2-aminoethoxydiphenyl borate (2-APB), prevented the senktide-induced growth in filopodia; demonstrating that NK3R-induced outgrowth of filopodia likely involves the release of intracellular calcium. Exposure of transfected CLU209 cells to senktide for 24 h led to further growth of filopodia and processes that extended 10-20 µm. A mathematical model, composed of a linear and population model was developed to account for the dynamics of filopodia growth during a timescale of minutes. The results suggest that the ligand-induced activation of NK3R affects early developmental processes by initiating filopodia formation that are a prerequisite for neuritogenesis.
authors.affiliation: Graduate Neuroscience Program and Department of Zoology and Physiology, University of Wyoming, Laramie, Wyoming, 82071.
authors.forename: Francis W
authors.initials: FW
authors.lastname: Flynn
authors.forename: Eli
authors.initials: E
authors.lastname: Kinney-Lang
authors.forename: Chelsea
authors.initials: C
authors.lastname: Hoekstra
authors.forename: Donald L
authors.initials: DL
authors.lastname: Pratt
authors.forename: Amit
authors.initials: A
authors.lastname: Thakar
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000502:physiology*; D007031:Hypothalamus / Q000196:embryology / Q000502:physiology*; D011554:Pseudopodia / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D018042:Receptors, Neurokinin-3 / Q000378:metabolism / Q000502:physiology*
chemical_list: D018042:Receptors, Neurokinin-3
keywords: NK3R; filopodia; senktide