Abstract
We studied the synthesis and release of nerve growth factor (NGF) in cultured rat iris with a two-site enzyme immunoassay by measuring the time course of NGF levels remaining in the iris and relased into the medium up to 72 h. For up to 3 h, the NGF levels in the iris did not change significantly. After that, they increased to a maximal level of 350 +/- 30 pg NGF/iris at 19 h, which is 200 times higher than the in vivo content. Between 20 and 72 h in culture, the NGF level decreased to 130 +/- 10 pg NGF/iris, whereas general protein synthesis did not change during that time period. Maximal rate of NGF production (203 pg NGF/h/iris) was seen between 9 and 12 h in culture. In the medium, NGF levels were first detectable after 6 h. Levels then increased with a time course similar to that seen within the iris, reaching a maximal level of 1,180 +/- 180 pg after 19 h in vitro, and then did not significantly change for up to 48 h. The NGF production of the densely sympathetically innervated dilator was three times higher than that of the predominantly cholinergically innervated sphincter. The NGF production was blocked by inhibitors of messenger RNA synthesis (actinomycin D) and of polyadenylation (9-beta-D- arabinofuranosyladenine) as well as by inhibitors of translation (cycloheximide). Monensin, which interferes with the transport of proteins through the Golgi apparatus, decreased NGF levels to 8-12% of controls in the medium, suggesting that the Golgi apparatus is involved in the intracellular processing of NGF.
Full Text
The Full Text of this article is available as a PDF (530.4 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ebendal T., Olson L., Seiger A., Hedlund K. O. Nerve growth factors in the rat iris. Nature. 1980 Jul 3;286(5768):25–28. doi: 10.1038/286025a0. [DOI] [PubMed] [Google Scholar]
- Ebendal T., Olson L., Seiger A. The level of nerve growth factor (NGF) as a function of innervation. A correlation radio-immunoassay and bioassay study of the rat iris. Exp Cell Res. 1983 Oct 15;148(2):311–317. doi: 10.1016/0014-4827(83)90155-6. [DOI] [PubMed] [Google Scholar]
- Gumbiner B., Kelly R. B. Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells. Cell. 1982 Jan;28(1):51–59. doi: 10.1016/0092-8674(82)90374-9. [DOI] [PubMed] [Google Scholar]
- Harper G. P., Al-Saffar A. M., Pearce F. L., Vernon C. A. The production of nerve growth factor in vitro by tissues of the mouse, rat, and embryonic chick. Dev Biol. 1980 Jun 15;77(2):379–390. doi: 10.1016/0012-1606(80)90482-0. [DOI] [PubMed] [Google Scholar]
- Harper G. P., Thoenen H. Target cells, biological effects, and mechanism of action of nerve growth factor and its antibodies. Annu Rev Pharmacol Toxicol. 1981;21:205–229. doi: 10.1146/annurev.pa.21.040181.001225. [DOI] [PubMed] [Google Scholar]
- Korsching S., Thoenen H. Nerve growth factor in sympathetic ganglia and corresponding target organs of the rat: correlation with density of sympathetic innervation. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3513–3516. doi: 10.1073/pnas.80.11.3513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Levi-Montalcini R., Angeletti P. U. Nerve growth factor. Physiol Rev. 1968 Jul;48(3):534–569. doi: 10.1152/physrev.1968.48.3.534. [DOI] [PubMed] [Google Scholar]
- Lubińska L. On axoplasmic flow. Int Rev Neurobiol. 1975;17:241–296. doi: 10.1016/s0074-7742(08)60211-1. [DOI] [PubMed] [Google Scholar]
- Malmfors T., Sachs C. Direct studies on the disappearance of the transmitter and changes in the uptake-storage mechanisms of degenerating adrenergic nerves. Acta Physiol Scand. 1965 Jul;64(3):211–223. doi: 10.1111/j.1748-1716.1965.tb04171.x. [DOI] [PubMed] [Google Scholar]
- Malmfors T. The adrenergic innervation of the eye as demonstrated by fluorescence microscopy. Acta Physiol Scand. 1965 Nov;65(3):259–267. doi: 10.1111/j.1748-1716.1965.tb04269.x. [DOI] [PubMed] [Google Scholar]
- Rose K. M., Leonard T. B., Carter T. H. Effects of adenine nucleosides on RNA synthesis in adenovirus-infected cells. 9-beta-D-Arabinofuranosyladenine as a selective inhibitor of RNA polyadenylation. Mol Pharmacol. 1982 Sep;22(2):517–523. [PubMed] [Google Scholar]
- Schwab M. E., Heumann R., Thoenen H. Communication between target organs and nerve cells: retrograde axonal transport and site of action of nerve growth factors. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):125–134. doi: 10.1101/sqb.1982.046.01.016. [DOI] [PubMed] [Google Scholar]
- Schwab M. E., Stöckel K., Thoenen H. Immunocytochemical localization of nerve growth factor (NGF) in the submandibular gland of adult mice by light and electron microscopy. Cell Tissue Res. 1976 Jun 28;169(3):289–299. doi: 10.1007/BF00219602. [DOI] [PubMed] [Google Scholar]
- Tartakoff A. M. Perturbation of vesicular traffic with the carboxylic ionophore monensin. Cell. 1983 Apr;32(4):1026–1028. doi: 10.1016/0092-8674(83)90286-6. [DOI] [PubMed] [Google Scholar]
- Tartakoff A. M. The confined function model of the Golgi complex: center for ordered processing of biosynthetic products of the rough endoplasmic reticulum. Int Rev Cytol. 1983;85:221–252. doi: 10.1016/S0074-7696(08)62374-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thoenen H., Barde Y. A. Physiology of nerve growth factor. Physiol Rev. 1980 Oct;60(4):1284–1335. doi: 10.1152/physrev.1980.60.4.1284. [DOI] [PubMed] [Google Scholar]
- Zyznar E. S. A rationale for the application of trasylol as a protease inhibitor in radioimmunoassay. Life Sci. 1981 Apr 27;28(17):1861–1866. doi: 10.1016/0024-3205(81)90291-5. [DOI] [PubMed] [Google Scholar]