Stigmoid bodies contain type I receptor proteins SorLA/LR11 and sortilin: new perspectives on their function.
Stigmoid bodies (SBs) are structures in the cytoplasm of neurons. SBs are mostly found in the hypothalamic region of the rat and contain a protein called huntingtin-associated protein 1 (HAP1). In a recent publication, large cytoplasmic structures were shown to be immunoreactive for a type I receptor called SorLA/LR11. By light microscopic analysis, these structures appeared similar to SBs in size and in brain regional and subcellular localization. To determine whether these large puncta correspond to HAP1-containing SBs, we used antibodies specific to various domains of the apolipoprotein receptor LR11 to perform immunocytochemistry in rat and mouse brain tissue. Transfection studies using HeLa cells were conducted to demonstrate the specificity of the antibodies. We found that, in both species, antibodies to the domain II (or VSP10 for vacuolar sorting protein 10 domain) of LR11 immunoreact with large cytoplasmic structures. Co-localization immunolabeling experiments in rat brain tissue sections and in neuron cultures showed that these LR11-immunoreactive structures correspond to HAP1-positive SBs. Electron microscopy was performed in rat hypothalamus and further demonstrated the presence of LR11 in SBs and its co-localization with HAP1. LR11-containing SBs were most abundant in the hypothalamus but were also found in many brainstem nuclei, thalamus, and hippocampus. Our data also show that sortilin, another transmembrane protein containing a VPS10 domain, localizes to large cytoplasmic puncta and is found in LR11-positive and Hap1-positive SBs in hypothalamic neuron cultures.
Gutekunst
Claire-Anne
CA
Department of Neurology, Emory University School of Medicine, Atlanta, Georgia 30322, USA. mguteku@emory.edu
Torre
Enrique R
ER
Sheng
Zejuan
Z
Yi
Hong
H
Coleman
Sarah H
SH
Riedel
I Björn
IB
Bujo
Hideaki
H
D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000033:anatomy & histology / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism / Q000648:ultrastructure; D005333:Fetus / Q000166:cytology; D005455:Fluorescent Antibody Technique; D006801:Humans; D026502:LDL-Receptor Related Proteins; D008562:Membrane Glycoproteins / Q000378:metabolism* / Q000502:physiology; D026901:Membrane Transport Proteins; D051379:Mice; D008854:Microscopy, Electron; D009419:Nerve Tissue Proteins / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D011973:Receptors, LDL / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D000099183:Sortilin
D033942:Adaptor Proteins, Vesicular Transport; D000911:Antibodies, Monoclonal; D026502:LDL-Receptor Related Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D011973:Receptors, LDL; D000099183:Sortilin; C096896:HAP1 protein, human; C490758:Hap1 protein, mouse; C490757:Hap1 protein, rat; C102582:SORL1 protein, human; C491168:Sorl1 protein, mouse; C491167:Sorl1 protein, rat