Characterization of hypothalamic subcellular particles containing luteinzing hormone releasing hormone and thyrotropin releasing hormone

A. Barnea, N. Ben Jonathan, J. C. Porter

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The 900g supernatant fluid prepared from male rat hypothalamic homogenates was fractionated by means of continuous sucrose density gradient centrifugation. Thyrotropin releasing hormone and luteinizing hormone releasing hormone in the gradient fractions were quantified by radioimmunoassays. TRH was associated with two populations of particles separable by means of nonequilibrium density centrifugation (100,000g for 30 min). However, after 'equilibrium' centrifugation (100,000xg for 180 min), a single peak of TRH was observed at 1.07 M sucrose. Hypo osmotic shock as well as treatment with 0.1% Triton X 100 or 0.1% deoxycholate (DOC) released TRH from both sets of particles. LRH, as TRH, was associated with two populations of particles which were separable by means of nonequilibrium density gradient centrifugation. After 'equilibrium' centrifugation, both sets of LRH containing particles banded at 1.27 M sucrose as a single symmetrical peak. Although 0.1% Triton X 100 released LRH from both populations of particles, hypo osmotic shock or 0.1% DOC released LRH only from the large LRH containing particles. The small LRH containing particles were resistant to hypo osmotic shock and to 0.1% DOC. Based on these criteria, it is concluded that in hypothalamic homogenates the TRH containing particles and the large LRH containing particles are synaptosomes. The small LRH containing particles may be of different cellular and/or subcellular origin.

Original languageEnglish (US)
Pages (from-to)477-484
Number of pages8
JournalJournal of Neurochemistry
Volume27
Issue number2
StatePublished - 1976

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Deoxycholic Acid
Thyrotropin-Releasing Hormone
Centrifugation
Hormones
Osmotic Pressure
Sucrose
Density Gradient Centrifugation
Octoxynol
Population
Synaptosomes
Gonadotropin-Releasing Hormone
Radioimmunoassay
Rats
Fluids

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Characterization of hypothalamic subcellular particles containing luteinzing hormone releasing hormone and thyrotropin releasing hormone. / Barnea, A.; Ben Jonathan, N.; Porter, J. C.

In: Journal of Neurochemistry, Vol. 27, No. 2, 1976, p. 477-484.

Research output: Contribution to journalArticle

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N2 - The 900g supernatant fluid prepared from male rat hypothalamic homogenates was fractionated by means of continuous sucrose density gradient centrifugation. Thyrotropin releasing hormone and luteinizing hormone releasing hormone in the gradient fractions were quantified by radioimmunoassays. TRH was associated with two populations of particles separable by means of nonequilibrium density centrifugation (100,000g for 30 min). However, after 'equilibrium' centrifugation (100,000xg for 180 min), a single peak of TRH was observed at 1.07 M sucrose. Hypo osmotic shock as well as treatment with 0.1% Triton X 100 or 0.1% deoxycholate (DOC) released TRH from both sets of particles. LRH, as TRH, was associated with two populations of particles which were separable by means of nonequilibrium density gradient centrifugation. After 'equilibrium' centrifugation, both sets of LRH containing particles banded at 1.27 M sucrose as a single symmetrical peak. Although 0.1% Triton X 100 released LRH from both populations of particles, hypo osmotic shock or 0.1% DOC released LRH only from the large LRH containing particles. The small LRH containing particles were resistant to hypo osmotic shock and to 0.1% DOC. Based on these criteria, it is concluded that in hypothalamic homogenates the TRH containing particles and the large LRH containing particles are synaptosomes. The small LRH containing particles may be of different cellular and/or subcellular origin.

AB - The 900g supernatant fluid prepared from male rat hypothalamic homogenates was fractionated by means of continuous sucrose density gradient centrifugation. Thyrotropin releasing hormone and luteinizing hormone releasing hormone in the gradient fractions were quantified by radioimmunoassays. TRH was associated with two populations of particles separable by means of nonequilibrium density centrifugation (100,000g for 30 min). However, after 'equilibrium' centrifugation (100,000xg for 180 min), a single peak of TRH was observed at 1.07 M sucrose. Hypo osmotic shock as well as treatment with 0.1% Triton X 100 or 0.1% deoxycholate (DOC) released TRH from both sets of particles. LRH, as TRH, was associated with two populations of particles which were separable by means of nonequilibrium density gradient centrifugation. After 'equilibrium' centrifugation, both sets of LRH containing particles banded at 1.27 M sucrose as a single symmetrical peak. Although 0.1% Triton X 100 released LRH from both populations of particles, hypo osmotic shock or 0.1% DOC released LRH only from the large LRH containing particles. The small LRH containing particles were resistant to hypo osmotic shock and to 0.1% DOC. Based on these criteria, it is concluded that in hypothalamic homogenates the TRH containing particles and the large LRH containing particles are synaptosomes. The small LRH containing particles may be of different cellular and/or subcellular origin.

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