Skip to main content
SpringerLink
Log in

Investigation of the Effects of PACAP on the Composition of Tear and Endolymph Proteins

  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Pituitary adenylate cyclase activating polypeptide (PACAP) is widely distributed in ocular tissues, including the lacrimal gland. PACAP has been shown to influence the activity of several exocrine glands, but its effects on the composition of the tear film are not known yet. Similarly, the presence of PACAP has already been shown in the inner ear, but it is not known whether PACAP influences the composition of the endolymph. The aim of the present study was to investigate whether systemic injection of PACAP has any modulatory effects on the protein composition of the tear film and endolymph using chip electrophoresis and mass spectrometry analysis. Tear and endolymph samples were collected from rats and chickens, respectively, at various time points after systemic injection of PACAP. Fluid samples were further processed for chip electrophoretic studies. No difference was found in the protein composition of the endolymph between control and PACAP-treated animals. In contrast, tear samples showed a marked difference after PACAP treatment. Proteins in the molecular range 50–70 kDa, which showed a different chip electropherogram profile in every PACAP-treated sample, were further analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. PACAP treatment induced a repression in certain keratins, while others were induced after PACAP injection. Furthermore, PACAP treatment decreased aldehyde dehydrogenase expression. The present study provides a base for further studies on the in vivo effects of PACAP on the composition of tear film. These investigations may have important clinical relevance because of the noninvasive sample collection, the correlation between tear proteins and ocular diseases, and the possible presence of biomarkers for both ophthalmological and systemic pathological conditions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  • Abu-Hamdan, M. D., Drescher, M. J., Ramakrishnan, N. A., et al. (2006). Pituitary adenylate cyclase activating polypeptide (PACAP) and its receptor (PAC1-R) in the cochlea: Evidence for specific transcript expression of PAC1-R splice variants in rat miscrodissected cochlear subfractions. Neuroscience, 140, 147–161.

    Article  PubMed  CAS  Google Scholar 

  • Babai, N., Atlasz, T., Tamas, A., Reglodi, D., Kiss, P., & Gabriel, R. (2005). Degree of damage compensation by various PACAP treatments in monosodium glutamate-induced retina degeneration. Neurotoxicity Research, 8, 227–233.

    Article  PubMed  CAS  Google Scholar 

  • Baker, G. R., Morton, M., Rajapaska, R. S., et al. (2006). Altered tear composition in smokers and patients with Graves ophthalmopathy. Archives of Ophthalmology, 124, 1451–1456.

    Article  PubMed  CAS  Google Scholar 

  • Borba, J. C., Henze, I. P., Silveira, M. S., et al. (2005). Pituitary adenylate cyclase activating polypeptide (PACAP) can act as determinant of the tyrosine hydoxylase phenotype of dopaminergic cells during retina development. Developmental Brain Research, 156, 193–201.

    Article  PubMed  CAS  Google Scholar 

  • Calvert, P. A., Heck, P. M., & Edwards, A. V. (1998). Autonomic control of submandibular protein secretion in the anaesthetized calf. Experimental Physiology, 83, 545–556.

    PubMed  CAS  Google Scholar 

  • Couloigner, V., Teixeira, M., Sterkers, O., Rask-Andersen, H., & Ferrary, E. (2004). The endolymphatic sac: its roles in the inner ear. Medical Science (Paris), 20, 304–310.

    Google Scholar 

  • De Souza, G. A., Godoy, L. M., & Mann, M. (2006). Identification of 491 proteins in the tear fluid proteome reveals a large number if proteases and protease inhibitors. Genome Biology, 7, R72.

    Article  PubMed  Google Scholar 

  • Drescher, M. J., Drescher, D. G., Khan, K. M., et al. (2006). Pituitary adenylate cyclase activating polypeptide (PACAP) and its receptor (PAC-1R) are positioned to modulate afferent signaling in the cochlea. Neuroscience, 142, 139–164.

    Article  PubMed  CAS  Google Scholar 

  • Eckert, R. L., & Green, H. (1984). Cloning of cDNAs specifying vitamin A-responsive human keratins. Proceedings of the National Academy of Sciences of the United States of America, 81, 4321–4325.

    Article  PubMed  CAS  Google Scholar 

  • Elsas, T., Uddman, R., & Sundler, F. (1996). Pituitary adenylate cyclase activating peptide-immunoreactive nerve fibers in the cat eye. Graefe's Archive for Clinical and Experimental Ophthalmology, 234, 573–580.

    Article  PubMed  CAS  Google Scholar 

  • Ferrary, E., Bernard, C., Teixeira, M., et al. (1996). Hormonal modulation of inner ear fluids. Acta Oto-laryngologica, 116, 244–247.

    Article  PubMed  CAS  Google Scholar 

  • Ferrary, E., & Sterkers, O. (1998). Mechanisms of endolymph secretion. Kidney International Supplement, 65, S98–S103.

    PubMed  CAS  Google Scholar 

  • Fukushima, K., Takeda, T., Kakigi, A., et al. (2005). Effects of lithium on endolymph homeostasis and experimentally induced endolymphatic hydrops. ORL Journal for Oto-rhino-laryngoly and its Related Specialties, 67, 282–288.

    CAS  Google Scholar 

  • Gilbard, J. P., Rossi, S. R., Heyda, K. G., & Dartt, D. A. (1990). Stimulation of tear secretion by topical agents that increase cyclic nucleotide levels. Investigative Ophthalmology in Visual Science, 31, 1381–1388.

    CAS  Google Scholar 

  • Grus, F. H., Podust, V. N., Bruns, K., et al. (2005). SELDI-TOF-MS ProteinChip array profiling of tears from patients with dry eye. Investigative Ophthalmology in Visual Science, 46, 863–876.

    Article  Google Scholar 

  • Hanner, P., Jennische, E., Lange, S., Lonnroth, I., & Wahlstrom, B. (2004). Increased antisecretory factor reduces vertigo in patients with Meniere’s disease: A pilot study. Hearing Research, 190, 31–36.

    Article  PubMed  CAS  Google Scholar 

  • Hannibal, J., & Fahrenkrug, J. (2004). Target areas innervated by PACAP-immunoreactive retinal ganglion cells. Cell & Tissue Research, 316, 99–113.

    Article  CAS  Google Scholar 

  • Hu, S., Loo, J. A., & Wong, D. T. (2006). Human body fluid proteome analysis. Proteomics, 6, 6326–6353.

    Article  PubMed  CAS  Google Scholar 

  • Jozsa, R., Somogyvari-Vigh, A., Reglodi, D., Hollosy, T., & Arimura, A. (2001). Distribution and daily variations of PACAP in the chicken brain. Peptides, 22, 1371–1377.

    Article  PubMed  CAS  Google Scholar 

  • Kamaishi, H., Endoh, T., & Suzuki, T. (2004). Multiple signal pathways coupling VIP and PACAP receptors to calcium channels in hamster submandibular ganglion neurons. Autonomic Neuroscience, 111, 15–26.

    Article  PubMed  CAS  Google Scholar 

  • Kapicioglu, Z., Kalyoncu, I. N., Deger, O., & Can, G. (1998). Effect of a somatostatin analogue (SMS 201-995) on tear secretion in rats. International Ophtalmology, 22, 43–45.

    Article  CAS  Google Scholar 

  • Kawano, H., Shimozono, M., Tono, T., Miyata, A., & Komune, S. (2001). Expression of pituitary adenylate cyclase activating polypeptide mRNA in the cochlea of rats. Molecular Brain Research, 94, 200–203.

    Article  PubMed  CAS  Google Scholar 

  • Kawasaki, S., Tanioka, H., Yamasaki, K., Yokoi, N., Komuro, A., & Kinoshita, S. (2006). Clusters of corneal epithelial cells reside ectopically in human conjunctival epithelium. Investigative Ophthalmology in Visual Science, 47, 1359–1367.

    Article  Google Scholar 

  • Kays, W. T., & Piatigorsky, J. (1997). Aldehyde dehydrogenase class 3 expression: identification of a cornea-preferred gene promoter in transgenic mice. Proceedings of the National Academy of Sciences of the United States of America, 94, 13594–13599.

    Article  PubMed  CAS  Google Scholar 

  • Kitano, H., Suzuki, M., Kitanishi, T., et al. (1999). Regulation of inner ear fluid in the rat by vasopressin. Neuroreport, 10, 1205–1207.

    Article  PubMed  CAS  Google Scholar 

  • Kovacs, I., Ludany, A., Koszegi, T., et al. (2005). Substance P released from sensory nerve endings influences tear secretion and goblet cell function in the rat. Neuropeptides, 39, 395–402.

    Article  PubMed  CAS  Google Scholar 

  • Kurpakus, M. A., Stock, E. L., & Jones, J. C. (1990). Expression of the 55-kD–64-kD corneal keratins in ocular surface epithelium. Investigative Ophthalmology in Visual Science, 31, 448–456.

    CAS  Google Scholar 

  • Kustos, I., Andrasfalvy, M., Kustos, T., Kocsis, B., & Kilar, F. (2005). Effect of iron restriction on outer membrane protein composition of Pseudomonas strains studied by conventional and microchip electrophoresis. Electrophoresis, 26, 3789–3795.

    Article  PubMed  CAS  Google Scholar 

  • Lebon, A., Seyer, D., Cosette, P., et al. (2006). Identification of proteins regulated by PACAP in PC12 cells by 2D gel electrophoresis coupled to mass spectrometry. Annals of the New York Academy of Sciences, 1070, 380–387.

    Article  PubMed  CAS  Google Scholar 

  • Li, N., Wang, N., Zheng, J., et al. (2005). Characterization of human tear proteome using multiple proteomic analysis techniques. Journal in Proteome Research, 4, 2052–2061.

    Article  CAS  Google Scholar 

  • Mirfendereski, S., Tobin, G., Hakanson, R., & Ekstrom, J. (1997). Pituitary adenylate cyclase activating peptide (PACAP) in salivary glands of the rat: origin, and secretory and vascular effects. Acta Physiologica Scandinavica, 160, 15–22.

    PubMed  CAS  Google Scholar 

  • Nagano, T., Nakamura, M., Nakata, K., et al. (2003). Effects of substance P and IGF-1 in corneal epithelial barrier function and wound healing in a rat model of neurotrophic keratopathy. Investigative Ophthalmology in Visual Science, 44, 3810–3815.

    Article  Google Scholar 

  • Nakatani, M., Seki, T., Shinohara, Y., et al. (2006). Pituitary adenylate cyclase activating polypeptide (PACAP) stimulates production of interleukin-6 in rat Muller cells. Peptides, 27, 1871–1876.

    Article  PubMed  CAS  Google Scholar 

  • Nilsson, S. F. (1994). PACAP-27 and PACAP-38: Vascular effects in the eye and some other tissues in the rabbit. European Journal of Pharmacology, 253, 17–25.

    Article  PubMed  CAS  Google Scholar 

  • Nilsson, S. F., De Neef, P., Robberecht, P., & Christophe, J. (1994). Characterization of ocular receptors for pituitary adenylate cyclase activating polypeptide (PACAP) and their coupling to adenylate cyclase. Experimental Eye Research, 58, 459–467.

    Article  PubMed  CAS  Google Scholar 

  • Pedersen, A. M., Dissing, S., Fahrenkrug, J., Hannibal, J., Reibel, J., & Nauntofte, B. (2000). Innervation pattern and Ca2+ signalling in labial salivary glands of healthy individuals and patients with primary Sjogren’s syndrome (pSS). Journal of Oral Pathology & Medicine, 29, 97–109.

    Article  CAS  Google Scholar 

  • Pei, Y., Reins, R. Y., & McDermott, A. M. (2006). Eldehyde dehydrogenase (ALDH) 3A1 expression by the human keratocyte and its repair phenotypes. Experimental Eye Research, 83, 1063–1073.

    Article  PubMed  CAS  Google Scholar 

  • Peterfi, Z., Kustos, I., Kilar, F., & Kocsis, B. (2007). Microfluidic chip analysis of outer membrane proteins responsible for serological cross-reaction between three Gram-negative bacteria: Proteus morganii O34, Escherichia coli O111 and Salmonella adelaide O35. Journal of Chromatography, 1155, 214–217.

    Article  PubMed  CAS  Google Scholar 

  • Porter, M. R., & Lane, E. B. (2003). Phenotypes, genotypes and their contribution to understanding keratin function. Trends in Genetics, 19, 278–285.

    Article  PubMed  CAS  Google Scholar 

  • Racz, B., Gallyas Jr., F., Kiss, P., et al. (2006). The neuroprotective effects of PACAP in monosodium glutamate-induced retinal lesion involves inhibition of proapoptotic signaling pathways. Regulatory Peptides, 137, 20–26.

    Article  PubMed  CAS  Google Scholar 

  • Racz, B., Gallyas Jr., F., Kiss, P., et al. (2007). Effects of pituitary adenylate cyclase activating polypeptide (PACAP) on the PKA-Bad-14-3-3 signaling pathway in glutamate-induced retinal injury in neonatal rats. Neurotoxicity Research, 12, 95–104.

    PubMed  CAS  Google Scholar 

  • Raphael, Y., & Altschuler, R. A. (2003). Structure and innervation of the cochlea. Brain Research Bulletin, 60, 397–422.

    Article  PubMed  Google Scholar 

  • Reglodi, D., Somogyvari-Vigh, A., Vigh, S., Kozicz, T., & Arimura, A. (2000). Delayed systemic administration of PACAP38 is neuroprotective in transient middle cerebral artery occlusion in the rat. Stroke, 31, 1411–1417.

    PubMed  CAS  Google Scholar 

  • Reglodi, D., Tamas, A., Somogyvári-Vigh, A., et al. (2002). Effects of pretreatment with PACAP on the infarct size and functional outcome in rat permanent focal cerebral ischemia. Peptides, 23, 2227–2234.

    Article  PubMed  CAS  Google Scholar 

  • Seki, T., Izumi, S., Shioda, S., Zhou, C. J., Arimura, A., & Koide, R. (2000b). Gene expression for PACAP receptor mRNA in the rat retina by in situ hybridization and in situ RT-PCR. Annals of the New York Academy of Sciences, 921, 366–369.

    Article  PubMed  CAS  Google Scholar 

  • Seki, T., Shioda, S., Izumi, S., Arimura, A., & Koide, R. (2000a). Electron microscopic observation of pituitary adenylate cyclase activating polypeptide (PACAP)-containing neurons in the rat retina. Peptides, 21, 109–113.

    Article  PubMed  CAS  Google Scholar 

  • Shioda, S., Ohtaki, H., Nakamachi, T., et al. (2006). Pleiotropic functions of PACAP in the CNS. Neuroprotection and neurodevelopment. Annals of the New York Academy of Sciences, 1070, 550–560.

    Article  PubMed  CAS  Google Scholar 

  • Shoge, K., Mishima, H. K., Saitoh, T., et al. (1999). Attenuation by PACAP of glutamate-induced neurotoxicity in cultured retinal neurons. Brain Research, 839, 66–73.

    Article  PubMed  CAS  Google Scholar 

  • Skakkabaek, M., Hannibal, J., & Fahrenkrug, J. (1999). Pituitary adenylate cyclase activating polypeptide (PACAP) in the rat mammary gland. Cell & Tissue Research, 298, 153–159.

    Article  Google Scholar 

  • Somogyvari-Vigh, A., & Reglodi, D. (2004). Pituitary adenylate cyclase activating polypeptide: A potential neuroprotective peptide. Review of Current Pharmaceutical Design, 10, 2861–2889.

    Article  CAS  Google Scholar 

  • Tanaka, K., & Smith, C. A. (1978). Structure of the chicken’s inner ear: SEM and TEM study. American Journal of Anatomy, 153, 251–271.

    Article  PubMed  CAS  Google Scholar 

  • Thalmann, R., & Thalmann, I. (1999). Source and role of endolymph macromolecules. Acta Oto-laryngologica, 119, 293–296.

    Article  PubMed  CAS  Google Scholar 

  • Tobin, G., Asztely, A., Edwards, A. V., Ekstrom, J., Hakanson, R., & Sundler, F. (1995). Presence and effects of pituitary adenylate cyclase activating peptide in the submandibular gland of the ferret. Neuroscience, 66, 227–235.

    Article  PubMed  CAS  Google Scholar 

  • Tomosugi, N., Kitagawa, K., Takahashi, N., Sugai, S., & Ishikawa, I. (2005). Diagnostic potential of tear proteomic patters in Sjogren’s syndrome. Journal of Proteome Research, 4, 820–825.

    Article  PubMed  CAS  Google Scholar 

  • Vaudry, D., Gonzalez, B. J., Basille, M., Yon, L., Fournier, A., & Vaudry, H. (2000). Pituitary adenylate cyclase activating polypeptide and its receptors: From structure to functions. Pharmacological Reviews, 52, 269–324.

    PubMed  CAS  Google Scholar 

  • Wang, Z. Y., Alm, P., & Hakanson, R. (1995). Distribution and effects of pituitary adenylate cyclase activating peptide in the rabbit eye. Neuroscience, 69, 297–308.

    Article  PubMed  CAS  Google Scholar 

  • Waschek, J. A. (2002). Multiple actions of pituitary adenylyl cyclase activating peptide in nervous system development and regeneration. Developmental Neuroscience, 24, 14–23.

    Article  PubMed  CAS  Google Scholar 

  • Yoshitomi, T., Yamaji, K., Ushikawa, H., & Ohnishi, Y. (2002). Effect of pituitary adenylate cyclase activating peptide on isolated rabbit iris sphincter and dilator muscles. Investigative Ophthalmology in Visual Science, 43, 780–783.

    Google Scholar 

Download references

Acknowledgments

This work was supported by the Hungarian Science Research Fund (OTKA T046589, F 67830, K 72592, K 68476 and F 048908) and ETT439/2006.

Author information

Authors and Affiliations

  1. Department of Ophthalmology, University of Pecs, Ifjusag u 13, 7624, Pecs, Hungary

    Valeria Gaal

  2. Department of Biochemistry and Medical Chemistry, University of Pecs, Szigeti u 12, 7624, Pecs, Hungary

    Laszlo Mark & Viktoria Nemeth

  3. Department of Anatomy, University of Pecs, Szigeti u 12, 7624, Pecs, Hungary

    Peter Kiss, Andrea Tamas, Andrea Lubics & Dora Reglodi

  4. Department of Medical Microbiology and Immunology, University of Pecs, Szigeti u 12, 7624, Pecs, Hungary

    Ildiko Kustos & Bela Kocsis

  5. Department of Oto-Rhino-Laryngology, University of Pecs, Munkacsy M u 2, 7621, Pecs, Hungary

    Adrienn Nemeth, Laszlo Lujber & Jozsef Pytel

  6. Department of Medical Chemistry, University of Szeged, Dom ter 8, 6720, Szeged, Hungary

    Gabor Toth

Authors
  1. Valeria Gaal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Laszlo Mark
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Peter Kiss
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ildiko Kustos
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Andrea Tamas
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bela Kocsis
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Andrea Lubics
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Viktoria Nemeth
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Adrienn Nemeth
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Laszlo Lujber
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Jozsef Pytel
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Gabor Toth
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Dora Reglodi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dora Reglodi.

Additional information

Valera Gaal and Laszlo Mark made equal contribution to the present work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gaal, V., Mark, L., Kiss, P. et al. Investigation of the Effects of PACAP on the Composition of Tear and Endolymph Proteins. J Mol Neurosci 36, 321–329 (2008). https://doi.org/10.1007/s12031-008-9067-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12031-008-9067-5

Keywords

Search

Navigation