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
Gespeichert in:
| Autor*in: |
Wibullaksanakul, Sunee [verfasserIn] Handelsman, David J. [verfasserIn] |
|---|
| Format: |
E-Artikel |
|---|
| Erschienen: |
Oxford, UK: Blackwell Publishing Ltd ; 1991 |
|---|
| Schlagwörter: |
|---|
| Umfang: |
Online-Ressource |
|---|
| Reproduktion: |
2006 ; Blackwell Publishing Journal Backfiles 1879-2005 |
|---|---|
| Übergeordnetes Werk: |
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 |
| Links: |
|---|
| DOI / URN: |
10.1111/j.1365-2826.1991.tb00261.x |
|---|
| Katalog-ID: |
NLEJ240220285 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLEJ240220285 | ||
| 003 | DE-627 | ||
| 005 | 20210707104605.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 120426s1991 xx |||||o 00| ||und c | ||
| 024 | 7 | |a 10.1111/j.1365-2826.1991.tb00261.x |2 doi | |
| 035 | |a (DE-627)NLEJ240220285 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 100 | 1 | |a Wibullaksanakul, Sunee |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems |
| 264 | 1 | |a Oxford, UK |b Blackwell Publishing Ltd |c 1991 | |
| 300 | |a Online-Ressource | ||
| 336 | |a nicht spezifiziert |b zzz |2 rdacontent | ||
| 337 | |a nicht spezifiziert |b z |2 rdamedia | ||
| 338 | |a nicht spezifiziert |b zu |2 rdacarrier | ||
| 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. | ||
| 533 | |d 2006 |f Blackwell Publishing Journal Backfiles 1879-2005 |7 |2006|||||||||| | ||
| 650 | 4 | |a gonadotropin-releasing hormone | |
| 700 | 1 | |a Handelsman, David J. |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i In |t Journal of neuroendocrinology |d Oxford [u.a.] : Wiley-Blackwell, 1989 |g 3(1991), 2, Seite 0 |h Online-Ressource |w (DE-627)NLEJ243926375 |w (DE-600)2007386-0 |x 1365-2826 |7 nnns |
| 773 | 1 | 8 | |g volume:3 |g year:1991 |g number:2 |g pages:0 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1111/j.1365-2826.1991.tb00261.x |q text/html |x Verlag |z Deutschlandweit zugänglich |3 Volltext |
| 912 | |a GBV_USEFLAG_U | ||
| 912 | |a ZDB-1-DJB | ||
| 912 | |a GBV_NL_ARTICLE | ||
| 951 | |a AR | ||
| 952 | |d 3 |j 1991 |e 2 |h 0 | ||
| author_variant |
s w sw d j h dj djh |
|---|---|
| matchkey_str |
article:13652826:1991----::yohlmcoaorpneesnhroeertomtoooia |
| hierarchy_sort_str |
1991 |
| publishDate |
1991 |
| allfields |
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 |
| spelling |
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 |
| allfields_unstemmed |
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 |
| allfieldsGer |
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 |
| allfieldsSound |
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 |
| source |
In Journal of neuroendocrinology 3(1991), 2, Seite 0 volume:3 year:1991 number:2 pages:0 |
| sourceStr |
In Journal of neuroendocrinology 3(1991), 2, Seite 0 volume:3 year:1991 number:2 pages:0 |
| format_phy_str_mv |
Article |
| institution |
findex.gbv.de |
| topic_facet |
gonadotropin-releasing hormone |
| isfreeaccess_bool |
false |
| container_title |
Journal of neuroendocrinology |
| authorswithroles_txt_mv |
Wibullaksanakul, Sunee @@aut@@ Handelsman, David J. @@aut@@ |
| publishDateDaySort_date |
1991-01-01T00:00:00Z |
| hierarchy_top_id |
NLEJ243926375 |
| id |
NLEJ240220285 |
| fullrecord |
<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">NLEJ240220285</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20210707104605.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">120426s1991 xx |||||o 00| ||und c</controlfield><datafield tag="024" ind1="7" ind2=" "><subfield code="a">10.1111/j.1365-2826.1991.tb00261.x</subfield><subfield code="2">doi</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)NLEJ240220285</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Wibullaksanakul, Sunee</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Oxford, UK</subfield><subfield code="b">Blackwell Publishing Ltd</subfield><subfield code="c">1991</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">Online-Ressource</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zzz</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">z</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zu</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="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.</subfield></datafield><datafield tag="533" ind1=" " ind2=" "><subfield code="d">2006</subfield><subfield code="f">Blackwell Publishing Journal Backfiles 1879-2005</subfield><subfield code="7">|2006||||||||||</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">gonadotropin-releasing hormone</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Handelsman, David J.</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">In</subfield><subfield code="t">Journal of neuroendocrinology</subfield><subfield code="d">Oxford [u.a.] : Wiley-Blackwell, 1989</subfield><subfield code="g">3(1991), 2, Seite 0</subfield><subfield code="h">Online-Ressource</subfield><subfield code="w">(DE-627)NLEJ243926375</subfield><subfield code="w">(DE-600)2007386-0</subfield><subfield code="x">1365-2826</subfield><subfield code="7">nnns</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:3</subfield><subfield code="g">year:1991</subfield><subfield code="g">number:2</subfield><subfield code="g">pages:0</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">http://dx.doi.org/10.1111/j.1365-2826.1991.tb00261.x</subfield><subfield code="q">text/html</subfield><subfield code="x">Verlag</subfield><subfield code="z">Deutschlandweit zugänglich</subfield><subfield code="3">Volltext</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ZDB-1-DJB</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_NL_ARTICLE</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">3</subfield><subfield code="j">1991</subfield><subfield code="e">2</subfield><subfield code="h">0</subfield></datafield></record></collection>
|
| series2 |
Blackwell Publishing Journal Backfiles 1879-2005 |
| author |
Wibullaksanakul, Sunee |
| spellingShingle |
Wibullaksanakul, Sunee misc gonadotropin-releasing hormone Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems |
| authorStr |
Wibullaksanakul, Sunee |
| ppnlink_with_tag_str_mv |
@@773@@(DE-627)NLEJ243926375 |
| format |
electronic Article |
| delete_txt_mv |
keep |
| author_role |
aut aut |
| collection |
NL |
| publishPlace |
Oxford, UK |
| remote_str |
true |
| illustrated |
Not Illustrated |
| issn |
1365-2826 |
| topic_title |
Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems gonadotropin-releasing hormone |
| publisher |
Blackwell Publishing Ltd |
| publisherStr |
Blackwell Publishing Ltd |
| topic |
misc gonadotropin-releasing hormone |
| topic_unstemmed |
misc gonadotropin-releasing hormone |
| topic_browse |
misc gonadotropin-releasing hormone |
| format_facet |
Elektronische Aufsätze Aufsätze Elektronische Ressource |
| format_main_str_mv |
Text Zeitschrift/Artikel |
| carriertype_str_mv |
zu |
| hierarchy_parent_title |
Journal of neuroendocrinology |
| hierarchy_parent_id |
NLEJ243926375 |
| hierarchy_top_title |
Journal of neuroendocrinology |
| isfreeaccess_txt |
false |
| familylinks_str_mv |
(DE-627)NLEJ243926375 (DE-600)2007386-0 |
| title |
Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems |
| ctrlnum |
(DE-627)NLEJ240220285 |
| title_full |
Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems |
| author_sort |
Wibullaksanakul, Sunee |
| journal |
Journal of neuroendocrinology |
| journalStr |
Journal of neuroendocrinology |
| isOA_bool |
false |
| recordtype |
marc |
| publishDateSort |
1991 |
| contenttype_str_mv |
zzz |
| container_start_page |
0 |
| author_browse |
Wibullaksanakul, Sunee Handelsman, David J. |
| container_volume |
3 |
| physical |
Online-Ressource |
| format_se |
Elektronische Aufsätze |
| author-letter |
Wibullaksanakul, Sunee |
| doi_str_mv |
10.1111/j.1365-2826.1991.tb00261.x |
| author2-role |
verfasserin |
| title_sort |
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. |
| collection_details |
GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE |
| container_issue |
2 |
| title_short |
Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems |
| url |
http://dx.doi.org/10.1111/j.1365-2826.1991.tb00261.x |
| remote_bool |
true |
| author2 |
Handelsman, David J. |
| author2Str |
Handelsman, David J. |
| ppnlink |
NLEJ243926375 |
| mediatype_str_mv |
z |
| isOA_txt |
false |
| hochschulschrift_bool |
false |
| doi_str |
10.1111/j.1365-2826.1991.tb00261.x |
| up_date |
2024-07-06T09:22:13.273Z |
| _version_ |
1803820972926566400 |
| fullrecord_marcxml |
<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">NLEJ240220285</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20210707104605.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">120426s1991 xx |||||o 00| ||und c</controlfield><datafield tag="024" ind1="7" ind2=" "><subfield code="a">10.1111/j.1365-2826.1991.tb00261.x</subfield><subfield code="2">doi</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)NLEJ240220285</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Wibullaksanakul, Sunee</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Hypothalamic Gonadotropin-Releasing Hormone Secretion: Methodological Aspects of in vitro Systems</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Oxford, UK</subfield><subfield code="b">Blackwell Publishing Ltd</subfield><subfield code="c">1991</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">Online-Ressource</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zzz</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">z</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zu</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="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.</subfield></datafield><datafield tag="533" ind1=" " ind2=" "><subfield code="d">2006</subfield><subfield code="f">Blackwell Publishing Journal Backfiles 1879-2005</subfield><subfield code="7">|2006||||||||||</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">gonadotropin-releasing hormone</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Handelsman, David J.</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">In</subfield><subfield code="t">Journal of neuroendocrinology</subfield><subfield code="d">Oxford [u.a.] : Wiley-Blackwell, 1989</subfield><subfield code="g">3(1991), 2, Seite 0</subfield><subfield code="h">Online-Ressource</subfield><subfield code="w">(DE-627)NLEJ243926375</subfield><subfield code="w">(DE-600)2007386-0</subfield><subfield code="x">1365-2826</subfield><subfield code="7">nnns</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:3</subfield><subfield code="g">year:1991</subfield><subfield code="g">number:2</subfield><subfield code="g">pages:0</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">http://dx.doi.org/10.1111/j.1365-2826.1991.tb00261.x</subfield><subfield code="q">text/html</subfield><subfield code="x">Verlag</subfield><subfield code="z">Deutschlandweit zugänglich</subfield><subfield code="3">Volltext</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ZDB-1-DJB</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_NL_ARTICLE</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">3</subfield><subfield code="j">1991</subfield><subfield code="e">2</subfield><subfield code="h">0</subfield></datafield></record></collection>
|
| score |
7.399392 |