Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems
The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including st...
Ausführliche Beschreibung
Autor*in: |
Wibullaksanakul, Sunee [verfasserIn] Handelsman, David J. [verfasserIn] |
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Oxford, UK: Blackwell Publishing Ltd ; 1991 |
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Online-Ressource |
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2006 ; Blackwell Publishing Journal Backfiles 1879-2005 |
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In: Journal of neuroendocrinology - Oxford [u.a.] : Wiley-Blackwell, 1989, 3(1991), 2, Seite 0 |
Übergeordnetes Werk: |
volume:3 ; year:1991 ; number:2 ; pages:0 |
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DOI / URN: |
10.1111/j.1365-2826.1991.tb00261.x |
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520 | |a The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including stability of baseline GnRH release and technical limitations or artifacts as well as for response to norepinephrine and naloxone, an opiate antagonist, which represent the two major physiological neuromodulators regulating the GnRH neuron. Baseline GnRH release rate per MBH was similar in both systems for at least 6 h, although owing to dilution of effluent samples by flow-through medium, the dynamic superfusion system required six MBH per chamber compared with the static incubation system which required only one MBH per chamber. Procedural losses of GnRH were low with either system without the need for proteolytic enzyme inhibitors and were mainly due to adhesion to plastic surfaces of chambers and tubings in the dynamic superfusion system. Mechanical agitation of chambers by either manual swinging or continuous shaking increased GnRH release rate. Addition of hypertonic KCl (final 50 mM) or equivalent NaCl or mannitol solution in a minimal volume directly into the tissue chambers, induced a non-specific GnRH release due to osmotic effects in the dynamic superfusion system. In contrast, the static incubation system, where hypothalamic tissue is exposed to the exact final concentration of solute by complete change of media, was more resistant to GnRH release by hypertonic stimulus. Thus, only in the static incubation system did 50 mM KCl concentrations cause GnRH release through depolarization alone. Using systems optimized to avoid technical artifacts, stimulation with naloxone demonstrated dose-dependent GnRH release in both systems whereas norepinephrine gave a clear dose-dependent GnRH release only in the static system.Thus both incubation systems produce stable and similar baseline GnRH release for at least 6 h. The static incubation system proved superior requiring fewer hypothalami, a shorter preincubation stabilization period, allowing more accurate concentration of additives and was less prone to mechanical and osmotic artifacts while preserving responsiveness to physiological stimuli. The static incubation system, being technically simpler, more robust and accurate, provided a more valid approach to the in vitro study of GnRH release and its regulation by neurochemicals. | ||
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10.1111/j.1365-2826.1991.tb00261.x doi (DE-627)NLEJ240220285 DE-627 ger DE-627 rakwb Wibullaksanakul, Sunee verfasserin aut Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems Oxford, UK Blackwell Publishing Ltd 1991 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including stability of baseline GnRH release and technical limitations or artifacts as well as for response to norepinephrine and naloxone, an opiate antagonist, which represent the two major physiological neuromodulators regulating the GnRH neuron. Baseline GnRH release rate per MBH was similar in both systems for at least 6 h, although owing to dilution of effluent samples by flow-through medium, the dynamic superfusion system required six MBH per chamber compared with the static incubation system which required only one MBH per chamber. Procedural losses of GnRH were low with either system without the need for proteolytic enzyme inhibitors and were mainly due to adhesion to plastic surfaces of chambers and tubings in the dynamic superfusion system. Mechanical agitation of chambers by either manual swinging or continuous shaking increased GnRH release rate. Addition of hypertonic KCl (final 50 mM) or equivalent NaCl or mannitol solution in a minimal volume directly into the tissue chambers, induced a non-specific GnRH release due to osmotic effects in the dynamic superfusion system. In contrast, the static incubation system, where hypothalamic tissue is exposed to the exact final concentration of solute by complete change of media, was more resistant to GnRH release by hypertonic stimulus. Thus, only in the static incubation system did 50 mM KCl concentrations cause GnRH release through depolarization alone. Using systems optimized to avoid technical artifacts, stimulation with naloxone demonstrated dose-dependent GnRH release in both systems whereas norepinephrine gave a clear dose-dependent GnRH release only in the static system.Thus both incubation systems produce stable and similar baseline GnRH release for at least 6 h. The static incubation system proved superior requiring fewer hypothalami, a shorter preincubation stabilization period, allowing more accurate concentration of additives and was less prone to mechanical and osmotic artifacts while preserving responsiveness to physiological stimuli. The static incubation system, being technically simpler, more robust and accurate, provided a more valid approach to the in vitro study of GnRH release and its regulation by neurochemicals. 2006 Blackwell Publishing Journal Backfiles 1879-2005 |2006|||||||||| gonadotropin-releasing hormone Handelsman, David J. verfasserin aut In Journal of neuroendocrinology Oxford [u.a.] : Wiley-Blackwell, 1989 3(1991), 2, Seite 0 Online-Ressource (DE-627)NLEJ243926375 (DE-600)2007386-0 1365-2826 nnns volume:3 year:1991 number:2 pages:0 http://dx.doi.org/10.1111/j.1365-2826.1991.tb00261.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 3 1991 2 0 |
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10.1111/j.1365-2826.1991.tb00261.x doi (DE-627)NLEJ240220285 DE-627 ger DE-627 rakwb Wibullaksanakul, Sunee verfasserin aut Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems Oxford, UK Blackwell Publishing Ltd 1991 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including stability of baseline GnRH release and technical limitations or artifacts as well as for response to norepinephrine and naloxone, an opiate antagonist, which represent the two major physiological neuromodulators regulating the GnRH neuron. Baseline GnRH release rate per MBH was similar in both systems for at least 6 h, although owing to dilution of effluent samples by flow-through medium, the dynamic superfusion system required six MBH per chamber compared with the static incubation system which required only one MBH per chamber. Procedural losses of GnRH were low with either system without the need for proteolytic enzyme inhibitors and were mainly due to adhesion to plastic surfaces of chambers and tubings in the dynamic superfusion system. Mechanical agitation of chambers by either manual swinging or continuous shaking increased GnRH release rate. Addition of hypertonic KCl (final 50 mM) or equivalent NaCl or mannitol solution in a minimal volume directly into the tissue chambers, induced a non-specific GnRH release due to osmotic effects in the dynamic superfusion system. In contrast, the static incubation system, where hypothalamic tissue is exposed to the exact final concentration of solute by complete change of media, was more resistant to GnRH release by hypertonic stimulus. Thus, only in the static incubation system did 50 mM KCl concentrations cause GnRH release through depolarization alone. Using systems optimized to avoid technical artifacts, stimulation with naloxone demonstrated dose-dependent GnRH release in both systems whereas norepinephrine gave a clear dose-dependent GnRH release only in the static system.Thus both incubation systems produce stable and similar baseline GnRH release for at least 6 h. The static incubation system proved superior requiring fewer hypothalami, a shorter preincubation stabilization period, allowing more accurate concentration of additives and was less prone to mechanical and osmotic artifacts while preserving responsiveness to physiological stimuli. The static incubation system, being technically simpler, more robust and accurate, provided a more valid approach to the in vitro study of GnRH release and its regulation by neurochemicals. 2006 Blackwell Publishing Journal Backfiles 1879-2005 |2006|||||||||| gonadotropin-releasing hormone Handelsman, David J. verfasserin aut In Journal of neuroendocrinology Oxford [u.a.] : Wiley-Blackwell, 1989 3(1991), 2, Seite 0 Online-Ressource (DE-627)NLEJ243926375 (DE-600)2007386-0 1365-2826 nnns volume:3 year:1991 number:2 pages:0 http://dx.doi.org/10.1111/j.1365-2826.1991.tb00261.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 3 1991 2 0 |
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10.1111/j.1365-2826.1991.tb00261.x doi (DE-627)NLEJ240220285 DE-627 ger DE-627 rakwb Wibullaksanakul, Sunee verfasserin aut Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems Oxford, UK Blackwell Publishing Ltd 1991 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including stability of baseline GnRH release and technical limitations or artifacts as well as for response to norepinephrine and naloxone, an opiate antagonist, which represent the two major physiological neuromodulators regulating the GnRH neuron. Baseline GnRH release rate per MBH was similar in both systems for at least 6 h, although owing to dilution of effluent samples by flow-through medium, the dynamic superfusion system required six MBH per chamber compared with the static incubation system which required only one MBH per chamber. Procedural losses of GnRH were low with either system without the need for proteolytic enzyme inhibitors and were mainly due to adhesion to plastic surfaces of chambers and tubings in the dynamic superfusion system. Mechanical agitation of chambers by either manual swinging or continuous shaking increased GnRH release rate. Addition of hypertonic KCl (final 50 mM) or equivalent NaCl or mannitol solution in a minimal volume directly into the tissue chambers, induced a non-specific GnRH release due to osmotic effects in the dynamic superfusion system. In contrast, the static incubation system, where hypothalamic tissue is exposed to the exact final concentration of solute by complete change of media, was more resistant to GnRH release by hypertonic stimulus. Thus, only in the static incubation system did 50 mM KCl concentrations cause GnRH release through depolarization alone. Using systems optimized to avoid technical artifacts, stimulation with naloxone demonstrated dose-dependent GnRH release in both systems whereas norepinephrine gave a clear dose-dependent GnRH release only in the static system.Thus both incubation systems produce stable and similar baseline GnRH release for at least 6 h. The static incubation system proved superior requiring fewer hypothalami, a shorter preincubation stabilization period, allowing more accurate concentration of additives and was less prone to mechanical and osmotic artifacts while preserving responsiveness to physiological stimuli. The static incubation system, being technically simpler, more robust and accurate, provided a more valid approach to the in vitro study of GnRH release and its regulation by neurochemicals. 2006 Blackwell Publishing Journal Backfiles 1879-2005 |2006|||||||||| gonadotropin-releasing hormone Handelsman, David J. verfasserin aut In Journal of neuroendocrinology Oxford [u.a.] : Wiley-Blackwell, 1989 3(1991), 2, Seite 0 Online-Ressource (DE-627)NLEJ243926375 (DE-600)2007386-0 1365-2826 nnns volume:3 year:1991 number:2 pages:0 http://dx.doi.org/10.1111/j.1365-2826.1991.tb00261.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 3 1991 2 0 |
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10.1111/j.1365-2826.1991.tb00261.x doi (DE-627)NLEJ240220285 DE-627 ger DE-627 rakwb Wibullaksanakul, Sunee verfasserin aut Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems Oxford, UK Blackwell Publishing Ltd 1991 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including stability of baseline GnRH release and technical limitations or artifacts as well as for response to norepinephrine and naloxone, an opiate antagonist, which represent the two major physiological neuromodulators regulating the GnRH neuron. Baseline GnRH release rate per MBH was similar in both systems for at least 6 h, although owing to dilution of effluent samples by flow-through medium, the dynamic superfusion system required six MBH per chamber compared with the static incubation system which required only one MBH per chamber. Procedural losses of GnRH were low with either system without the need for proteolytic enzyme inhibitors and were mainly due to adhesion to plastic surfaces of chambers and tubings in the dynamic superfusion system. Mechanical agitation of chambers by either manual swinging or continuous shaking increased GnRH release rate. Addition of hypertonic KCl (final 50 mM) or equivalent NaCl or mannitol solution in a minimal volume directly into the tissue chambers, induced a non-specific GnRH release due to osmotic effects in the dynamic superfusion system. In contrast, the static incubation system, where hypothalamic tissue is exposed to the exact final concentration of solute by complete change of media, was more resistant to GnRH release by hypertonic stimulus. Thus, only in the static incubation system did 50 mM KCl concentrations cause GnRH release through depolarization alone. Using systems optimized to avoid technical artifacts, stimulation with naloxone demonstrated dose-dependent GnRH release in both systems whereas norepinephrine gave a clear dose-dependent GnRH release only in the static system.Thus both incubation systems produce stable and similar baseline GnRH release for at least 6 h. The static incubation system proved superior requiring fewer hypothalami, a shorter preincubation stabilization period, allowing more accurate concentration of additives and was less prone to mechanical and osmotic artifacts while preserving responsiveness to physiological stimuli. The static incubation system, being technically simpler, more robust and accurate, provided a more valid approach to the in vitro study of GnRH release and its regulation by neurochemicals. 2006 Blackwell Publishing Journal Backfiles 1879-2005 |2006|||||||||| gonadotropin-releasing hormone Handelsman, David J. verfasserin aut In Journal of neuroendocrinology Oxford [u.a.] : Wiley-Blackwell, 1989 3(1991), 2, Seite 0 Online-Ressource (DE-627)NLEJ243926375 (DE-600)2007386-0 1365-2826 nnns volume:3 year:1991 number:2 pages:0 http://dx.doi.org/10.1111/j.1365-2826.1991.tb00261.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 3 1991 2 0 |
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10.1111/j.1365-2826.1991.tb00261.x doi (DE-627)NLEJ240220285 DE-627 ger DE-627 rakwb Wibullaksanakul, Sunee verfasserin aut Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems Oxford, UK Blackwell Publishing Ltd 1991 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including stability of baseline GnRH release and technical limitations or artifacts as well as for response to norepinephrine and naloxone, an opiate antagonist, which represent the two major physiological neuromodulators regulating the GnRH neuron. Baseline GnRH release rate per MBH was similar in both systems for at least 6 h, although owing to dilution of effluent samples by flow-through medium, the dynamic superfusion system required six MBH per chamber compared with the static incubation system which required only one MBH per chamber. Procedural losses of GnRH were low with either system without the need for proteolytic enzyme inhibitors and were mainly due to adhesion to plastic surfaces of chambers and tubings in the dynamic superfusion system. Mechanical agitation of chambers by either manual swinging or continuous shaking increased GnRH release rate. Addition of hypertonic KCl (final 50 mM) or equivalent NaCl or mannitol solution in a minimal volume directly into the tissue chambers, induced a non-specific GnRH release due to osmotic effects in the dynamic superfusion system. In contrast, the static incubation system, where hypothalamic tissue is exposed to the exact final concentration of solute by complete change of media, was more resistant to GnRH release by hypertonic stimulus. Thus, only in the static incubation system did 50 mM KCl concentrations cause GnRH release through depolarization alone. Using systems optimized to avoid technical artifacts, stimulation with naloxone demonstrated dose-dependent GnRH release in both systems whereas norepinephrine gave a clear dose-dependent GnRH release only in the static system.Thus both incubation systems produce stable and similar baseline GnRH release for at least 6 h. The static incubation system proved superior requiring fewer hypothalami, a shorter preincubation stabilization period, allowing more accurate concentration of additives and was less prone to mechanical and osmotic artifacts while preserving responsiveness to physiological stimuli. The static incubation system, being technically simpler, more robust and accurate, provided a more valid approach to the in vitro study of GnRH release and its regulation by neurochemicals. 2006 Blackwell Publishing Journal Backfiles 1879-2005 |2006|||||||||| gonadotropin-releasing hormone Handelsman, David J. verfasserin aut In Journal of neuroendocrinology Oxford [u.a.] : Wiley-Blackwell, 1989 3(1991), 2, Seite 0 Online-Ressource (DE-627)NLEJ243926375 (DE-600)2007386-0 1365-2826 nnns volume:3 year:1991 number:2 pages:0 http://dx.doi.org/10.1111/j.1365-2826.1991.tb00261.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 3 1991 2 0 |
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Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems |
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Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems |
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Wibullaksanakul, Sunee |
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Journal of neuroendocrinology |
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1991 |
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Wibullaksanakul, Sunee Handelsman, David J. |
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Wibullaksanakul, Sunee |
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10.1111/j.1365-2826.1991.tb00261.x |
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hypothalamic gonadotropin-releasing hormone secretion: methodological aspects of in vitro systems |
title_auth |
Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems |
abstract |
The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including stability of baseline GnRH release and technical limitations or artifacts as well as for response to norepinephrine and naloxone, an opiate antagonist, which represent the two major physiological neuromodulators regulating the GnRH neuron. Baseline GnRH release rate per MBH was similar in both systems for at least 6 h, although owing to dilution of effluent samples by flow-through medium, the dynamic superfusion system required six MBH per chamber compared with the static incubation system which required only one MBH per chamber. Procedural losses of GnRH were low with either system without the need for proteolytic enzyme inhibitors and were mainly due to adhesion to plastic surfaces of chambers and tubings in the dynamic superfusion system. Mechanical agitation of chambers by either manual swinging or continuous shaking increased GnRH release rate. Addition of hypertonic KCl (final 50 mM) or equivalent NaCl or mannitol solution in a minimal volume directly into the tissue chambers, induced a non-specific GnRH release due to osmotic effects in the dynamic superfusion system. In contrast, the static incubation system, where hypothalamic tissue is exposed to the exact final concentration of solute by complete change of media, was more resistant to GnRH release by hypertonic stimulus. Thus, only in the static incubation system did 50 mM KCl concentrations cause GnRH release through depolarization alone. Using systems optimized to avoid technical artifacts, stimulation with naloxone demonstrated dose-dependent GnRH release in both systems whereas norepinephrine gave a clear dose-dependent GnRH release only in the static system.Thus both incubation systems produce stable and similar baseline GnRH release for at least 6 h. The static incubation system proved superior requiring fewer hypothalami, a shorter preincubation stabilization period, allowing more accurate concentration of additives and was less prone to mechanical and osmotic artifacts while preserving responsiveness to physiological stimuli. The static incubation system, being technically simpler, more robust and accurate, provided a more valid approach to the in vitro study of GnRH release and its regulation by neurochemicals. |
abstractGer |
The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including stability of baseline GnRH release and technical limitations or artifacts as well as for response to norepinephrine and naloxone, an opiate antagonist, which represent the two major physiological neuromodulators regulating the GnRH neuron. Baseline GnRH release rate per MBH was similar in both systems for at least 6 h, although owing to dilution of effluent samples by flow-through medium, the dynamic superfusion system required six MBH per chamber compared with the static incubation system which required only one MBH per chamber. Procedural losses of GnRH were low with either system without the need for proteolytic enzyme inhibitors and were mainly due to adhesion to plastic surfaces of chambers and tubings in the dynamic superfusion system. Mechanical agitation of chambers by either manual swinging or continuous shaking increased GnRH release rate. Addition of hypertonic KCl (final 50 mM) or equivalent NaCl or mannitol solution in a minimal volume directly into the tissue chambers, induced a non-specific GnRH release due to osmotic effects in the dynamic superfusion system. In contrast, the static incubation system, where hypothalamic tissue is exposed to the exact final concentration of solute by complete change of media, was more resistant to GnRH release by hypertonic stimulus. Thus, only in the static incubation system did 50 mM KCl concentrations cause GnRH release through depolarization alone. Using systems optimized to avoid technical artifacts, stimulation with naloxone demonstrated dose-dependent GnRH release in both systems whereas norepinephrine gave a clear dose-dependent GnRH release only in the static system.Thus both incubation systems produce stable and similar baseline GnRH release for at least 6 h. The static incubation system proved superior requiring fewer hypothalami, a shorter preincubation stabilization period, allowing more accurate concentration of additives and was less prone to mechanical and osmotic artifacts while preserving responsiveness to physiological stimuli. The static incubation system, being technically simpler, more robust and accurate, provided a more valid approach to the in vitro study of GnRH release and its regulation by neurochemicals. |
abstract_unstemmed |
The regulation of gonadotropin-releasing hormone (GnRH) secretion from the medial basal hypothalamus (MBH) of adult male rats has been studied using two types of in vitro methods, the dynamic superfusion and static incubation systems. We compared both systems for methodological features including stability of baseline GnRH release and technical limitations or artifacts as well as for response to norepinephrine and naloxone, an opiate antagonist, which represent the two major physiological neuromodulators regulating the GnRH neuron. Baseline GnRH release rate per MBH was similar in both systems for at least 6 h, although owing to dilution of effluent samples by flow-through medium, the dynamic superfusion system required six MBH per chamber compared with the static incubation system which required only one MBH per chamber. Procedural losses of GnRH were low with either system without the need for proteolytic enzyme inhibitors and were mainly due to adhesion to plastic surfaces of chambers and tubings in the dynamic superfusion system. Mechanical agitation of chambers by either manual swinging or continuous shaking increased GnRH release rate. Addition of hypertonic KCl (final 50 mM) or equivalent NaCl or mannitol solution in a minimal volume directly into the tissue chambers, induced a non-specific GnRH release due to osmotic effects in the dynamic superfusion system. In contrast, the static incubation system, where hypothalamic tissue is exposed to the exact final concentration of solute by complete change of media, was more resistant to GnRH release by hypertonic stimulus. Thus, only in the static incubation system did 50 mM KCl concentrations cause GnRH release through depolarization alone. Using systems optimized to avoid technical artifacts, stimulation with naloxone demonstrated dose-dependent GnRH release in both systems whereas norepinephrine gave a clear dose-dependent GnRH release only in the static system.Thus both incubation systems produce stable and similar baseline GnRH release for at least 6 h. The static incubation system proved superior requiring fewer hypothalami, a shorter preincubation stabilization period, allowing more accurate concentration of additives and was less prone to mechanical and osmotic artifacts while preserving responsiveness to physiological stimuli. The static incubation system, being technically simpler, more robust and accurate, provided a more valid approach to the in vitro study of GnRH release and its regulation by neurochemicals. |
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Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems |
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