A subset of chicken statoacoustic ganglion neurites are repelled by Slit1 and Slit2.
Mechanosensory hair cells in the chicken inner ear are innervated by bipolar afferent neurons of the statoacoustic ganglion (SAG). During development, individual SAG neurons project their peripheral process to only one of eight distinct sensory organs. These neuronal subtypes may respond differently to guidance cues as they explore the periphery in search of their target. Previous gene expression data suggested that Slit repellants might channel SAG neurites into the sensory primordia, based on the presence of robo transcripts in the neurons and the confinement of slit transcripts to the flanks of the prosensory domains. This led to the prediction that excess Slit proteins would impede the outgrowth of SAG neurites. As predicted, axonal projections to the primordium of the anterior crista were reduced 2-3 days after electroporation of either slit1 or slit2 expression plasmids into the anterior pole of the otocyst on embryonic day 3 (E3). The posterior crista afferents, which normally grow through and adjacent to slit expression domains as they are navigating towards the posterior pole of the otocyst, did not show Slit responsiveness when similarly challenged by ectopic delivery of slit to their targets. The sensitivity to ectopic Slits shown by the anterior crista afferents was more the exception than the rule: responsiveness to Slits was not observed when the entire E4 SAG was challenged with Slits for 40 h in vitro. The corona of neurites emanating from SAG explants was unaffected by the presence of purified human Slit1 and Slit2 in the culture medium. Reduced axon outgrowth from E8 olfactory bulbs cultured under similar conditions for 24 h confirmed bioactivity of purified human Slits on chicken neurons. In summary, differential sensitivity to Slit repellents may influence the directional outgrowth of otic axons toward either the anterior or posterior otocyst.
Department of Biological Sciences and Purdue University Center for Cancer Research, Purdue University, 915 W State St., West Lafayette, IN 47907-1392, USA. Electronic address: abattisti@medinst.com.
Andrea C
AC
Battisti
Department of Biological Sciences and Purdue University Center for Cancer Research, Purdue University, 915 W State St., West Lafayette, IN 47907-1392, USA. Electronic address: kfantetti@gmail.com.
Kristen N
KN
Fantetti
Department of Biological Sciences and Purdue University Center for Cancer Research, Purdue University, 915 W State St., West Lafayette, IN 47907-1392, USA. Electronic address: bamoyers@umich.edu.
Belle A
BA
Moyers
Department of Biological Sciences and Purdue University Center for Cancer Research, Purdue University, 915 W State St., West Lafayette, IN 47907-1392, USA. Electronic address: dfekete@purdue.edu.
Donna M
DM
Fekete
D000818:Animals; D030801:Animals, Genetically Modified; D030161:Avian Proteins / Q000235:genetics / Q000502:physiology*; D002642:Chick Embryo; D007758:Ear, Inner / Q000196:embryology / Q000294:innervation; D018274:Electroporation; D005724:Ganglia / Q000196:embryology / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000502:physiology*; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016501:Neurites / Q000145:classification / Q000502:physiology; D009475:Neurons, Afferent / Q000502:physiology*; D011971:Receptors, Immunologic / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D000097905:Roundabout Proteins; D000098552:Slit Homolog 2 Protein
D030161:Avian Proteins; D036341:Intercellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins; D011971:Receptors, Immunologic; D011994:Recombinant Proteins; D000097905:Roundabout Proteins; D000098552:Slit Homolog 2 Protein; C492289:SLIT1 protein, human