title: Immunocytochemical evidence of Tulp1-dependent outer segment protein transport pathways in photoreceptor cells.
abstract: Tulp1 is a protein of unknown function exclusive to rod and cone photoreceptor cells. Mutations in the gene cause autosomal recessive retinitis pigmentosa in humans and photoreceptor degeneration in mice. In tulp1-/- mice, rod and cone opsins are mislocalized, and rhodopsin-bearing extracellular vesicles accumulate around the inner segment, indicating that Tulp1 is involved in protein transport from the inner segment to the outer segment. To investigate this further, we sought to define which outer segment transport pathways are Tulp1-dependent. We used immunohistochemistry to examine the localization of outer segment proteins in tulp1-/- photoreceptors, prior to retinal degeneration. We also surveyed the condition of inner segment organelles and rhodopsin transport machinery proteins. Herein, we show that guanylate cyclase 1 and guanylate cyclase activating proteins 1 and 2 are mislocalized in the absence of Tulp1. Furthermore, arrestin does not translocate to the outer segment in response to light stimulation. Additionally, data from the tulp1-/- retina adds to the understanding of peripheral membrane protein transport, indicating that rhodopsin kinase and transducin do not co-transport in rhodopsin carrier vesicles and phosphodiesterase does not co-transport in guanylate cyclase carrier vesicles. These data implicate Tulp1 in the transport of selective integral membrane outer segment proteins and their associated proteins, specifically, the opsin and guanylate cyclase carrier pathways. The exact role of Tulp1 in outer segment protein transport remains elusive. However, without Tulp1, two rhodopsin transport machinery proteins exhibit abnormal distribution, Rab8 and Rab11, suggesting a role for Tulp1 in vesicular docking and fusion at the plasma membrane near the connecting cilium.
authors.affiliation: Department of Ophthalmic Research, i31, Cole Eye Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA.
authors.forename: Gregory H
authors.initials: GH
authors.lastname: Grossman
authors.forename: Rao F
authors.initials: RF
authors.lastname: Watson
authors.forename: Gayle J T
authors.initials: GJ
authors.lastname: Pauer
authors.forename: Kathryn
authors.initials: K
authors.lastname: Bollinger
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Hagstrom
mesh_terms: D000221:Adaptation, Ocular; D000818:Animals; D019393:Arrestin / Q000378:metabolism; D005136:Eye Proteins / Q000378:metabolism* / Q000502:physiology; D019084:Fluorescent Antibody Technique, Indirect; D051606:G-Protein-Coupled Receptor Kinase 1 / Q000378:metabolism; D006162:Guanylate Cyclase / Q000378:metabolism; D051604:Guanylate Cyclase-Activating Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D021381:Protein Transport; D011956:Receptors, Cell Surface / Q000378:metabolism; D055214:Retinal Photoreceptor Cell Outer Segment / Q000378:metabolism*; D015293:Transducin / Q000378:metabolism
chemical_list: D019393:Arrestin; D005136:Eye Proteins; D051604:Guanylate Cyclase-Activating Proteins; D011956:Receptors, Cell Surface; C491353:Tulp1 protein, mouse; C110715:guanylate cyclase 1; D051606:G-Protein-Coupled Receptor Kinase 1; D015293:Transducin; D006162:Guanylate Cyclase