Effect of host plant and immune challenge on the levels of chemosensory and odorant-binding proteins in caterpillar salivary glands.
More than half of the proteome from mandibular glands in caterpillars is represented by chemosensory proteins. Based on sequence similarity, these proteins are putative transporters of ligands to gustatory receptors in sensory organs of insects. We sought to determine whether these proteins are inducible by comparing, both qualitatively and quantitatively, the salivary (mandibular and labial) proteomes from caterpillars (Vanessa cardui) reared on different plants and artificial diet containing either bacteria or bacterial cell-walls. We included a treatment where the caterpillars were switched from feeding on artificial diet to plant material at some point in their development. Additionally, we evaluated the degree of overlap between the proteomes in the hemolymph-filled coelom and salivary glands of caterpillars reared on plant material. We found that the quality and quantity of the identified proteins differed clearly between hemolymph-filled coelome, labial and mandibular glands. Our results indicated that even after molting and two-day feeding on a new diet, protein production is affected by the previous food source used by the caterpillar. Candidate proteins involved in chemosensory perception by insects were detected: three chemosensory (CSPs) and two odorant-binding proteins (OBPs). Using the relative amounts of these proteins across tissues and treatments as criteria for their classification, we detected hemolymph- and mandibular gland-specific CSPs and observed that their levels were affected by caterpillar diet. Moreover, we could compare the protein and transcript levels across tissues and treatment for at least one CSP and one OBP. Therefore, we have identified specific isoforms for testing the role of CSPs and OBPs in plant and pathogen recognition. We detected catalase, immune-related protein and serine proteases and their inhibitors in high relative levels in the mandibular glands in comparison to the labial glands. These findings suggest that the mandibular glands of caterpillars may play an important role protecting the caterpillar from oxidative stress, pathogens and aiding in digestion. Contamination with hemolymph proteins during dissection of salivary glands from caterpillars may occur but it is not substantial since the proteomes from hemolymph, mandibular and labial glands were easily discriminated from each other by principal component analysis of proteomic data.
Stockholm University, Department of Zoology Ecology, Svante Arrheniusväg 18 B, 106 91, Stockholm, Sweden. Electronic address: maria.celorio@zoologi.su.se.
Maria de la Paz
Mde L
Celorio-Mancera
Karolinska Institute, Department of Medicine, Solna, Stockholm, Sweden. Electronic address: jimmy.ytterberg@ki.se.
A Jimmy
AJ
Ytterberg
Karolinska Institute, Division of Physiological Chemistry I, Department of Medical Biochemistry and Biophysics, Scheeles väg 2, S-171 77, Stockholm, Sweden; Science for Life Laboratory, Stockholm, Sweden. Electronic address: dorothea.rutishauser@ki.se.
Dorothea
D
Rutishauser
Stockholm University, Department of Zoology Ecology, Svante Arrheniusväg 18 B, 106 91, Stockholm, Sweden. Electronic address: niklas.janz@zoologi.su.se.
Niklas
N
Janz
Karolinska Institute, Division of Physiological Chemistry I, Department of Medical Biochemistry and Biophysics, Scheeles väg 2, S-171 77, Stockholm, Sweden; Science for Life Laboratory, Stockholm, Sweden. Electronic address: roman.zubarev@ki.se.
Roman A
RA
Zubarev
D000818:Animals; D002080:Butterflies / Q000254:growth & development / Q000378:metabolism*; D005260:Female; D006458:Hemolymph / Q000378:metabolism; D019476:Insect Proteins / Q000378:metabolism*; D007814:Larva / Q000378:metabolism; D008297:Male; D020543:Proteome / Q000378:metabolism*; D018035:Receptors, Odorant / Q000378:metabolism*; D012469:Salivary Glands / Q000378:metabolism
D019476:Insect Proteins; D020543:Proteome; D018035:Receptors, Odorant; C045648:odorant-binding protein
Chemosensory protein; Hemolymph; Insect immunity; Mandibular salivary gland; Odorant-binding protein; Proteomics