Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Aug;78(8):5216–5220. doi: 10.1073/pnas.78.8.5216

A new hypothalamic substance, and not luteinizing hormone-releasing hormone, is detected immunocytochemically by antibody to luteinizing hormone-releasing hormone.

L A Sternberger, J L Greenwald, D Hock, K Elger, W G Forssmann
PMCID: PMC320378  PMID: 7029530

Abstract

Adjacent paraffin sections of rat hypothalami fixed in Bouin's fluid were treated either with buffer or with luteinizing hormone-releasing hormone (LHRH) before immunocytochemical staining with anti-LHRH. Upon buffer pretreatment, pituitary gonadotrophs were unstained and hypothalamic fibers were stained. Upon LHRH pretreatment, pituitary gonadotrophs were stained (receptor reaction) and hypothalamic fibers were unstained. Extension of washes and use of series of neutralizing antisera between LHRH application and immunocytochemical staining, as well as the absence of inhibiting concentrations of LHRH in the later washes and neutralizing antisera removed from the sections, excluded the possibility that the disappearance of visualization of hypothalamic fibers was due to blockage of anti-LHRH in immunocytochemical staining. The results suggested that LHRH removed from the sections an immunocytochemically stainable but as yet unknown analog of LHRH and replaced it with LHRH, which in turn became lost during subsequent immunocytochemical processing. This idea was confirmed by the isolation by high-pressure liquid chromatography of a peak, distinct from LHRH, upon treatment of hypothalami with LHRH. It is suggested that the new substance may be carrier-held and that this substance, rather than LHRH, is normally detected by immunocytochemistry with anti-LHRH. Added LHRH binds not only to high-affinity pituitary receptors but also to low-affinity hypothalamic carriers.

Full text

PDF
5216

Images in this article

5217Image
on p.5217
5217Image
on p.5217
5217Image
on p.5217
5217Image
on p.5217

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams J. C. Technical considerations on the use of horseradish peroxidase as a neuronal marker. Neuroscience. 1977;2(1):141–145. doi: 10.1016/0306-4522(77)90074-4. [DOI] [PubMed] [Google Scholar]
  2. Alumets J., Håkanson R., Lundqvist G., Sundler F., Thorell J. Ontogeny and ultrastructure of somatostatin and calcitonin cells in the thyroid gland of the rat. Cell Tissue Res. 1980;206(2):193–201. doi: 10.1007/BF00232763. [DOI] [PubMed] [Google Scholar]
  3. Bloch B., Bugnon C., Fellmann D., Lenys D., Gouget A. Neurons of the rat hypothalamus reactive with antisera against endorphins, ACTH, MSH and beta-LPH. Cell Tissue Res. 1979;204(1):1–15. doi: 10.1007/BF00235160. [DOI] [PubMed] [Google Scholar]
  4. Boer H. H., Schot L. P., Roubos E. W., ter Maat A., Lodder J. C., Reichelt D., Swaab D. F. ACTH-like immunoreactivity in two electronically coupled giant neurons in the pond snail Lymnaea stagnalis. Cell Tissue Res. 1979 Nov;202(2):231–240. doi: 10.1007/BF00232237. [DOI] [PubMed] [Google Scholar]
  5. Buffa R., Chayvialle J. A., Fontana P., Usellini L., Capella C., Solcia E. Parafollicular cells of rabbit thryoid store both calcitonin and somatostatin and resemble gut D cells ultrastructurally. Histochemistry. 1979 Aug;62(3):281–288. doi: 10.1007/BF00508356. [DOI] [PubMed] [Google Scholar]
  6. Fritsch H. A., Van Noorden S., Pearse A. G. Calcitonin-like immunochemical staining in the alimentary tract of Ciona intestinalis L. Cell Tissue Res. 1980;205(3):439–444. doi: 10.1007/BF00232284. [DOI] [PubMed] [Google Scholar]
  7. Galmiche J. P., Chayvialle J. A., Dubois P. M., David L., Descos F., Paulin C., Ducastelle T., Colin R., Geffroy Y. Calcitonin-producing pancreatic somatostatinoma. Gastroenterology. 1980 Jun;78(6):1577–1583. [PubMed] [Google Scholar]
  8. Goldsmith P. C., Ganong W. F. Ultrastructural localization of luteinizing hormone-releasing hormone in the median eminence of the rat. Brain Res. 1975 Oct 31;97(2):181–193. doi: 10.1016/0006-8993(75)90444-8. [DOI] [PubMed] [Google Scholar]
  9. Grube D. Alpha-endorphin-like immunoreactivity in plasma cells of the canine colonic mucosa. Histochemistry. 1980;69(2):157–160. doi: 10.1007/BF00533132. [DOI] [PubMed] [Google Scholar]
  10. Grube D., Voigt K. H., Weber E. Pancreatic glucagon cells contain endorphin-like immunoreactivity. Histochemistry. 1978 Dec 28;59(1):75–79. doi: 10.1007/BF00506480. [DOI] [PubMed] [Google Scholar]
  11. Joseph S. A., Sternberger L. A. The unlabeled antibody method. Contrasting color staining of beta-lipotropin and ACTH-associated hypothalamic peptides without antibody removal. J Histochem Cytochem. 1979 Nov;27(11):1430–1437. doi: 10.1177/27.11.92499. [DOI] [PubMed] [Google Scholar]
  12. Krieger D. T., Liotta A. S. Pituitary hormones in brain: where, how, and why? Science. 1979 Jul 27;205(4404):366–372. doi: 10.1126/science.221983. [DOI] [PubMed] [Google Scholar]
  13. Larsson L. I. Distribution of ACTH-like immunoreactivity in rat brain and gastrointestinal tract. Histochemistry. 1978 Apr 4;55(3):225–233. doi: 10.1007/BF00495761. [DOI] [PubMed] [Google Scholar]
  14. Larsson L. I., Polak J. M., Buffa R., Sundler F., Solcia E. On the immunocytochemical localization of the vasoactive intestinal polypeptide. J Histochem Cytochem. 1979 May;27(5):936–938. doi: 10.1177/27.5.479555. [DOI] [PubMed] [Google Scholar]
  15. Larsson L. I., Rehfeld J. F. Localization and molecular heterogeneity of cholecystokinin in the central and peripheral nervous system. Brain Res. 1979 Apr 13;165(2):201–218. doi: 10.1016/0006-8993(79)90554-7. [DOI] [PubMed] [Google Scholar]
  16. Leonardelli J., Tramu G. Immunoreactivity for beta-endorphin in LH-RH neurons of the fetal human hypothalamus. Cell Tissue Res. 1979;203(2):201–207. doi: 10.1007/BF00237234. [DOI] [PubMed] [Google Scholar]
  17. Li C. H., Chung D. Isolation and structure of an untriakontapeptide with opiate activity from camel pituitary glands. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1145–1148. doi: 10.1073/pnas.73.4.1145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lorén I., Alumets J., Håkanson R., Sundler F. Immunoreactive pancreatic polypeptide (PP) occurs in the central and peripheral nervous system: preliminary immunocytochemical observations. Cell Tissue Res. 1979 Aug;200(2):179–186. doi: 10.1007/BF00236410. [DOI] [PubMed] [Google Scholar]
  19. Martin R., Frösch D., Weber E., Voigt K. H. Met-enkephaline-like immunoreactivity in a cephalopod neurohemal organ. Neurosci Lett. 1979 Dec;15(2-3):253–257. doi: 10.1016/0304-3940(79)96122-6. [DOI] [PubMed] [Google Scholar]
  20. Nozaki M., Kobayashi H. Distribution of LHRH-like substance in the vertebrate brain as revealed by immunohistochemistry. Arch Histol Jpn. 1979 Jul;42(3):201–219. doi: 10.1679/aohc1950.42.201. [DOI] [PubMed] [Google Scholar]
  21. Salih H., Murthy G. S., Friesen H. G. Stability of hormone receptors with fixation: implications for immunocytochemical localization of receptors. Endocrinology. 1979 Jul;105(1):21–26. doi: 10.1210/endo-105-1-21. [DOI] [PubMed] [Google Scholar]
  22. Scharrer B. Peptidergic neurons: facts and trends. Gen Comp Endocrinol. 1978 Jan;34(1):50–62. doi: 10.1016/0016-6480(78)90243-5. [DOI] [PubMed] [Google Scholar]
  23. Sternberger L. A., Hardy P. H., Jr, Cuculis J. J., Meyer H. G. The unlabeled antibody enzyme method of immunohistochemistry: preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem. 1970 May;18(5):315–333. doi: 10.1177/18.5.315. [DOI] [PubMed] [Google Scholar]
  24. Sternberger L. A., Petrali J. P., Joseph S. A., Meyer H. G., Mills K. R. Specificity of the immunocytochemical luteinizing hormone-releasing hormone receptor reaction. Endocrinology. 1978 Jan;102(1):63–73. doi: 10.1210/endo-102-1-63. [DOI] [PubMed] [Google Scholar]
  25. Sternberger L. A., Petrali J. P. Quantitative immunocytochemistry of pituitary receptors for luteinizing hormone-releasing hormone. Cell Tissue Res. 1975 Sep 17;162(2):141–176. doi: 10.1007/BF00209204. [DOI] [PubMed] [Google Scholar]
  26. Sternberger L. A., Petrali J. P. The unlabeled antibody enzyme method. Attempted use of peroxidase-conjugated antigen as the third layer in the technique. J Histochem Cytochem. 1977 Sep;25(9):1036–1042. doi: 10.1177/25.9.333018. [DOI] [PubMed] [Google Scholar]
  27. Tougard C., Tixier-Vidal A., Avrameas S. Comparison between peroxidase-conjugated antigen or antibody and peroxidase-anti-peroxidase complex in a postembedding procedure. J Histochem Cytochem. 1979 Dec;27(12):1630–1633. doi: 10.1177/27.12.391996. [DOI] [PubMed] [Google Scholar]
  28. Watkins W. B. Presence of adrenocorticotropin and beta-endorphin immunoreactivities in the magnocellular neurosecretory system of the rat hypothalamus. Cell Tissue Res. 1980;207(1):65–80. doi: 10.1007/BF00239330. [DOI] [PubMed] [Google Scholar]
  29. Wolf G. A supposed immunohistochemical differentiation between neurophysin and its putative precursor. Acta Histochem Suppl. 1980;22:225–230. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES