Calpain-5 Expression in the Retina Localizes to Photoreceptor Synapses

Kellie A Schaefer; Marcus A Toral; Gabriel Velez; Allison J Cox; Sheila A Baker; Nicholas C Borcherding; Diana F Colgan; Vimala Bondada; Charles B Mashburn; Chen-Guang Yu; James W Geddes; Stephen H Tsang; Alexander G Bassuk; Vinit B Mahajan

doi:10.1167/iovs.15-18680

Back

Calpain-5 Expression in the Retina Localizes to Photoreceptor Synapses

Journal article

Open access

Peer reviewed

Calpain-5 Expression in the Retina Localizes to Photoreceptor Synapses

Kellie A Schaefer, Marcus A Toral, Gabriel Velez, Allison J Cox, Sheila A Baker, Nicholas C Borcherding, Diana F Colgan, Vimala Bondada, Charles B Mashburn, Chen-Guang Yu, …

Investigative ophthalmology & visual science, Vol.57(6), pp.2509-2521

05/01/2016

DOI: 10.1167/iovs.15-18680

PMCID: PMC4868102

PMID: 27152965

Files and links (1)

url

https://doi.org/10.1167/iovs.15-18680View

Published (Version of record) Open Access

Abstract

We characterize calpain-5 (CAPN5) expression in retinal and neuronal subcellular compartments. CAPN5 gene variants were classified using the exome variant server, and RNA-sequencing was used to compare expression of CAPN5 mRNA in the mouse and human retina and in retinoblastoma cells. Expression of CAPN5 protein was ascertained in humans and mice in silico, in mouse retina by immunohistochemistry, and in neuronal cancer cell lines and fractionated central nervous system tissue extracts by Western analysis with eight antibodies targeting different CAPN5 regions. Most CAPN5 genetic variation occurs outside its protease core; and searches of cancer and epilepsy/autism genetic databases found no variants similar to hyperactivating retinal disease alleles. The mouse retina expressed one transcript for CAPN5 plus those of nine other calpains, similar to the human retina. In Y79 retinoblastoma cells, the level of CAPN5 transcript was very low. Immunohistochemistry detected CAPN5 expression in the inner and outer nuclear layers and at synapses in the outer plexiform layer. Western analysis of fractionated retinal extracts confirmed CAPN5 synapse localization. Western blots of fractionated brain neuronal extracts revealed distinct subcellular patterns and the potential presence of autoproteolytic CAPN5 domains. CAPN5 is moderately expressed in the retina and, despite higher expression in other tissues, hyperactive disease mutants of CAPN5 only manifest as eye disease. At the cellular level, CAPN5 is expressed in several different functional compartments. CAPN5 localization at the photoreceptor synapse and with mitochondria explains the neural circuitry phenotype in human CAPN5 disease alleles.

Immunohistochemistry

Retina - metabolism

Retinoblastoma - genetics

Humans

Gene Expression Regulation, Neoplastic

Retinal Neoplasms - pathology

Male

Calpain - genetics

Retinoblastoma - pathology

Synapses - metabolism

Cattle

Neoplasms, Experimental

Female

Photoreceptor Cells - pathology

Retinal Neoplasms - metabolism

Tumor Cells, Cultured

Retinoblastoma - metabolism

Blotting, Western

Animals

Photoreceptor Cells - metabolism

Calpain - biosynthesis

RNA, Neoplasm - genetics

Mice

Retina - pathology

Retinal Neoplasms - genetics

Details

Title: Subtitle: Calpain-5 Expression in the Retina Localizes to Photoreceptor Synapses
Creators: Kellie A Schaefer - Omics Laboratory, University of Iowa, Iowa City, Iowa, United States 2Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City, Iowa, United States
Marcus A Toral - Omics Laboratory, University of Iowa, Iowa City, Iowa, United States 2Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City, Iowa, United States 3Medical Scientist Training Program, University of Iowa, Iowa City, Iowa, United States
Gabriel Velez - Omics Laboratory, University of Iowa, Iowa City, Iowa, United States 2Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City, Iowa, United States 3Medical Scientist Training Program, University of Iowa, Iowa City, Iowa, United States
Allison J Cox - Department of Pediatrics, University of Iowa, Iowa City, Iowa, United States
Sheila A Baker - Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City, Iowa, United States 5Department of Biochemistry, University of Iowa, Iowa City, Iowa, United States
Nicholas C Borcherding - Omics Laboratory, University of Iowa, Iowa City, Iowa, United States 3Medical Scientist Training Program, University of Iowa, Iowa City, Iowa, United States
Diana F Colgan - Omics Laboratory, University of Iowa, Iowa City, Iowa, United States 2Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City, Iowa, United States
Vimala Bondada - Spinal Cord and Brain Injury Research Center, University of Kentucky, Lexington, Kentucky, United States
Charles B Mashburn - Spinal Cord and Brain Injury Research Center, University of Kentucky, Lexington, Kentucky, United States
Chen-Guang Yu - Spinal Cord and Brain Injury Research Center, University of Kentucky, Lexington, Kentucky, United States
James W Geddes - Spinal Cord and Brain Injury Research Center, University of Kentucky, Lexington, Kentucky, United States
Stephen H Tsang - Barbara & Donald Jonas Stem Cell Laboratory, and Bernard & Shirlee Brown Glaucoma Laboratory, Department of Pathology & Cell Biology, Institute of Human Nutrition, College of Physicians and Surgeons, Columbia University, New York, New York, United States
Alexander G Bassuk - Department of Pediatrics, University of Iowa, Iowa City, Iowa, United States 9Neurology, University of Iowa, Iowa City, Iowa, United States
Vinit B Mahajan - Omics Laboratory, University of Iowa, Iowa City, Iowa, United States 2Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City, Iowa, United States
Resource Type: Journal article
Publication Details: Investigative ophthalmology & visual science, Vol.57(6), pp.2509-2521
DOI: 10.1167/iovs.15-18680
PMID: 27152965
PMCID: PMC4868102
NLM abbreviation: Invest Ophthalmol Vis Sci
ISSN: 0146-0404
eISSN: 1552-5783
Publisher: United States
Grant note: R01 EY020542 / NEI NIH HHS R01 EY025225 / NEI NIH HHS T32 GM008629 / NIGMS NIH HHS R01 EY024698 / NEI NIH HHS T32 GM007337 / NIGMS NIH HHS K08 EY020530 / NEI NIH HHS R01 EY024665 / NEI NIH HHS F30 EY027986 / NEI NIH HHS F31 EY026789 / NEI NIH HHS R21 AG050437 / NIA NIH HHS
Language: English
Date published: 05/01/2016
Academic Unit: Dermatology; Neurology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Biochemistry and Molecular Biology; Neurology (Pediatrics); University College Courses; Internal Medicine; Ophthalmology and Visual Sciences
Record Identifier: 9984020615702771

Metrics

27 Record Views

31 Times Cited - Web of Science