Distribution and biosynthesis of LH-RH and structurally related peptides

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dc.contributor.advisorMillar, Robert Pen_ZA
dc.contributor.authorDutlow, Clive Men_ZA
dc.date.accessioned2018-02-01T13:29:50Z
dc.date.available2018-02-01T13:29:50Z
dc.date.issued1988en_ZA
dc.description.abstractBiologically active peptides are a class of molecules which can, in general, be grouped into families of structurally related moieties which have a wide somatic distribution and which subserve multiple physiological roles. Thus, the releasing factors of the hypothalamus or their structural analogues may carry out multiple endocrine, paracrine, or neurotransmitter functions outside the hypothalamus. This thesis describes the distribution and biosynthesis of Luteinizing Hormone-Releasing Hormone (LH-RH) and structurally related peptides in mammalian tissues and tumours. Two strategies were employed in this investigation: (a) Immunological screening of tissues for suitable sources of LH-RH and the partial characterization of the LH-RH immunoreactive peptides of selected tissues; (b) Complementary oligonucleotide screening of mRNA from certain tissues, cDNA and genomic libraries as well as genomic DNA restriction endonuclease fragments to establish the nature of the LH-RH precursor and related molecules. Acetic acid extracts of nineteen rat tissues were assayed in serial dilution for LH-RH immunoreactivity (LH-RH-IR). An antibody directed to the middle of the LH-RH sequence was used in these screening radioimmunoassays. LH-RH-IR was found in extrahypothalamic brain areas, in gastroenteropancreatic tissues, in the retina, the submandibular gland, the thyroid and the testes. In view of a putative role of endogeonous LH-RH-like immunoreactive peptides in intra-testicular regulation, the molecular nature of the LH-RH-like material detected in rat testes was investigated. Acetic acid extracts of adult rat testes were partially purified by LH-RH-immunoaffinity chromatography. On Sephadex G-100 this material separated into four major peaks of >100K, ~32K, ~5K and <4K daltons. The <4K peak of LH-RH-IR eluted differently on Sephadex G-25 and analytical reverse phase HPLC than did synthetic hypothalamic LH-RH decapeptide. Antibodies directed at the C-terminus of LH-RH gave higher estimates of LH-RH-IR for all the chromatographically separated testicular polypeptides than did N-terminally and middle directed antisera. The small testicular LH-RH-like peptides displaced radiolabelled LH-RH agonist from rat pituitary membranes more effectively than did the larger immunoreactive molecules. Monkey, pig and dog testes as well as dog Sertoli cell tumours were extracted with acetic acid and shown to contain LH-RH immunoreactive material similar to that of the rat. Human seminal plasma was also investigated as a source of LH-RH-IR. When pooled seminal plasma from azoospermic males was fractionated by gel filtration some fractions had significant amounts of the rat testicular type of LH-RH.en_ZA
dc.identifier.apacitationDutlow, C. M. (1988). <i>Distribution and biosynthesis of LH-RH and structurally related peptides</i>. (Thesis). University of Cape Town ,Faculty of Health Sciences ,Division of Chemical Pathology. Retrieved from http://hdl.handle.net/11427/27200en_ZA
dc.identifier.chicagocitationDutlow, Clive M. <i>"Distribution and biosynthesis of LH-RH and structurally related peptides."</i> Thesis., University of Cape Town ,Faculty of Health Sciences ,Division of Chemical Pathology, 1988. http://hdl.handle.net/11427/27200en_ZA
dc.identifier.citationDutlow, C. 1988. Distribution and biosynthesis of LH-RH and structurally related peptides. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Dutlow, Clive M AB - Biologically active peptides are a class of molecules which can, in general, be grouped into families of structurally related moieties which have a wide somatic distribution and which subserve multiple physiological roles. Thus, the releasing factors of the hypothalamus or their structural analogues may carry out multiple endocrine, paracrine, or neurotransmitter functions outside the hypothalamus. This thesis describes the distribution and biosynthesis of Luteinizing Hormone-Releasing Hormone (LH-RH) and structurally related peptides in mammalian tissues and tumours. Two strategies were employed in this investigation: (a) Immunological screening of tissues for suitable sources of LH-RH and the partial characterization of the LH-RH immunoreactive peptides of selected tissues; (b) Complementary oligonucleotide screening of mRNA from certain tissues, cDNA and genomic libraries as well as genomic DNA restriction endonuclease fragments to establish the nature of the LH-RH precursor and related molecules. Acetic acid extracts of nineteen rat tissues were assayed in serial dilution for LH-RH immunoreactivity (LH-RH-IR). An antibody directed to the middle of the LH-RH sequence was used in these screening radioimmunoassays. LH-RH-IR was found in extrahypothalamic brain areas, in gastroenteropancreatic tissues, in the retina, the submandibular gland, the thyroid and the testes. In view of a putative role of endogeonous LH-RH-like immunoreactive peptides in intra-testicular regulation, the molecular nature of the LH-RH-like material detected in rat testes was investigated. Acetic acid extracts of adult rat testes were partially purified by LH-RH-immunoaffinity chromatography. On Sephadex G-100 this material separated into four major peaks of >100K, ~32K, ~5K and <4K daltons. The <4K peak of LH-RH-IR eluted differently on Sephadex G-25 and analytical reverse phase HPLC than did synthetic hypothalamic LH-RH decapeptide. Antibodies directed at the C-terminus of LH-RH gave higher estimates of LH-RH-IR for all the chromatographically separated testicular polypeptides than did N-terminally and middle directed antisera. The small testicular LH-RH-like peptides displaced radiolabelled LH-RH agonist from rat pituitary membranes more effectively than did the larger immunoreactive molecules. Monkey, pig and dog testes as well as dog Sertoli cell tumours were extracted with acetic acid and shown to contain LH-RH immunoreactive material similar to that of the rat. Human seminal plasma was also investigated as a source of LH-RH-IR. When pooled seminal plasma from azoospermic males was fractionated by gel filtration some fractions had significant amounts of the rat testicular type of LH-RH. DA - 1988 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 1988 T1 - Distribution and biosynthesis of LH-RH and structurally related peptides TI - Distribution and biosynthesis of LH-RH and structurally related peptides UR - http://hdl.handle.net/11427/27200 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/27200
dc.identifier.vancouvercitationDutlow CM. Distribution and biosynthesis of LH-RH and structurally related peptides. [Thesis]. University of Cape Town ,Faculty of Health Sciences ,Division of Chemical Pathology, 1988 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/27200en_ZA
dc.language.isoengen_ZA
dc.publisher.departmentDivision of Chemical Pathologyen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.subject.otherLH - biosynthesisen_ZA
dc.subject.otherPeptides - biosynthesisen_ZA
dc.titleDistribution and biosynthesis of LH-RH and structurally related peptidesen_ZA
dc.typeDoctoral Thesis
dc.type.qualificationlevelDoctoral
dc.type.qualificationnamePhDen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceThesisen_ZA
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