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Luteinizing Hormone-Releasing Hormone Quantified in Tissues and Slice Explant Cultures of Postnatal Rat Hypothalami
LH-releasing hormone (LHRH) peptide from postnatal rat preoptic area (POA)/hypothalamic tissues in vivo and slice explant cultures maintained in vitro was quantitated using an enzyme-linked immunosorbant assay. Moreover, messenger RNA (mRNA) copy number was calculated in LHRH neurons maintained in c...
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Published in: | Endocrinology (Philadelphia) 1999-02, Vol.140 (2), p.791-799 |
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Main Authors: | , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | LH-releasing hormone (LHRH) peptide from postnatal rat preoptic area
(POA)/hypothalamic tissues in vivo and slice explant
cultures maintained in vitro was quantitated using an
enzyme-linked immunosorbant assay. Moreover, messenger RNA (mRNA) copy
number was calculated in LHRH neurons maintained in culture using
in situ hybridization histochemistry with
autoradiographic film analysis. POA/hypothalami from postnatal day 5–6
pups averaged 1250 pg of LHRH, with approximately 28% of peptide
residing within rostral tissues where most LHRH perikarya reside.
Explant cultures maintained 18 days in vitro contained
30.4–92.0 pg/slice with a whole animal total of 244.8 pg. Considering
cell numbers in vivo and in vitro, LHRH
neurons in whole animal produce 1.0 pg of LHRH/cell, whereas those in
culture average 2.0 pg/cell. Furthermore, LHRH mRNA copies/cell in
organotypic culture was estimated conservatively at 1410 copies/cell, a
relatively high number. This work shows that, compared with whole
animal, cultures have substantial LHRH stores, indicating maturation of
synthetic activity and/or formation of new terminals in
vitro. High LHRH mRNA copy number also suggests a high rate of
peptide biosynthesis. Our analysis, demonstrating the dynamic potential
of LHRH neurons, suggests that subtle changes in LHRH mRNA expression
in all cells or a subpopulation can dramatically alter the LHRH system
biosynthetic capacity. |
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ISSN: | 0013-7227 1945-7170 |
DOI: | 10.1210/endo.140.2.6534 |