Subcellular Localization of Immunoreactive Thyroid-Stimulating Hormone in the Rat Hypothalamus

DeVito, William J.; Spearman, Terry N.; Connors, John M.; Hedge, George A.

doi:10.1159/000124488

Cited by 10 publications

(6 citation statements)

References 9 publications

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“…Tissues from 10 rats were homogenized with 10 strokes of a motor-driven teflon-glass homogenizer in 10 vol of ice-cold ho mogenizing medium (0.3 M sucrose, 10 mM 4-(2-hydroxyelhyl)-l-piperazineethanesulfonic acid KOH (HEPES KOH: pH 7.4), 2 mM EDTA and 0.2 mM EGTA) as previously described [7). Tis sue homogenates were centrifuged at 150 gfor 10 min at 4 °C, and the supernatants were layered onto 15 ml continuous sucrose gra dients (0.3-2.0 M) in 16 x 102 mm polyallomer centrifuge tubes.…”

Section: Subcellular Fractionationmentioning

confidence: 99%

Immunoreactive Prolactin in the Rat Hypothalamus: In vitro Release and Subcellular Localization

DeVito¹,

Connors²,

Hedge³

1987

Neuroendocrinology

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Immunocytochemical studies have identified immunoreactive prolactin (IR-PRL) in the hypothalamus and other areas of the rat brain. However, neither the release of IR-PRL from the hypothalamus nor its subcellular localization have been demonstrated. In this study, the release of IR-PRL from hypothalami obtained from female rats was examined using hypothalamic units incubated in vitro in Krebs-Ringer bicarbonate-glucose buffer. Hypothalamic tissue spontaneously released IR-PRL, and this release was increased by depolarizing concentrations of potassium by a calcium-dependent mechanism. Hypothalamic IR-PRL was also released from hypothalamic tissue obtained from hypophysectomized rats (14 days). The subcellular localization of IR-PRL was investigated using equilibrium-density centrifugation. Tissue homogenates from intact or hypophysectomized rats were centrifuged at 150 g at 4 °C for 10 min, and the supernatants were layered onto continuous sucrose gradients (1.00–1.27 g/ml) and centrifuged at 100,000 g (max.) for 16 h. IR-PRL in pituitary supernatants showed a high equilibrium-density peak with a modal density of 1.23 g/ml. Fractionation of the supernatant from ventral or dorsal hypothalamic tissue resulted in two high-equilibrium density peaks, a primary peak with a modal density of 1.23 g/ml and a smaller peak with a modal density of 1.10 g/ml. Both high-density peaks were maintained in tissue obtained from hypophysectomized rats and were disrupted by homogenization in hypo-osmotic medium. Together, these data suggest that hypothalamic IR-PRL is stored in membrane-bound particles which have densities similar to those of secretory granules and is released by a calcium-dependent mechanism when the tissue is depolarized.

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Section: Subcellular Fractionationmentioning

confidence: 99%

Immunoreactive Prolactin in the Rat Hypothalamus: In vitro Release and Subcellular Localization

DeVito¹,

Connors²,

Hedge³

1987

Neuroendocrinology

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“…This region is juxtaposed to the anterior pituitary that produces TSH in response to hypothalamic TRH. Furthermore, TSH has been reported to be expressed in the hypothalamus ( DeVito et al, 1986 ; Hojvat et al, 1983 ). It is therefore possible that a yet uncharacterized central TSH–TSHR feedback circuit may directly regulate the hypothalamic–pituitary–thyroid axis, thought solely to be controlled by thyroid hormones.…”

Section: Discussionmentioning

confidence: 99%

Brain atlas for glycoprotein hormone receptors at single-transcript level

Ryu

Гумерова

Korkmaz

et al. 2022

eLife

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There is increasing evidence that anterior pituitary hormones, traditionally thought to have unitary functions in regulating single endocrine targets, act on multiple somatic tissues, such as bone, fat, and liver. There is also emerging evidence for anterior pituitary hormone action on brain receptors in mediating central neural and peripheral somatic functions. Here, we have created the most comprehensive neuroanatomical atlas on the expression of TSHR, LHCGR and FSHR. We have used RNAscope, a technology that allows the detection of mRNA at single-transcript level, together with protein level validation, to document Tshr expression in 173 and Fshr expression in 353 brain regions, nuclei and sub-nuclei identified using the Atlas for the Mouse Brain in Stereotaxic Coordinates. We also identified Lhcgr transcripts in 401 brain regions, nuclei and sub-nuclei. Complementarily, we used ViewRNA, another single-transcript detection technology, to establish the expression of FSHR in human brain samples, where transcripts were co-localized in MALAT1 positive neurons. In addition, we show high expression for all three receptors in the ventricular region-with yet unknown functions. Intriguingly, Tshr and Fshr expression in the ependymal layer of the third ventricle was similar to that of the thyroid follicular cells and testicular Sertoli cells, respectively. In contrast, Fshr was localized to NeuN-positive neurons in the granular layer of the dentate gyrus in murine and human brain-both are Alzheimer's disease vulnerable regions. Our atlas thus provides a vital resource for scientists to explore the link between the stimulation or inactivation of brain glycoprotein hormone receptors on somatic function. New actionable pathways for human disease may be unmasked through further studies.

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“…This region is juxtaposed to the anterior pituitary that produces TSH in response to hypothalamic thyrotropin–releasing hormone (TRH). Furthermore, TSH has been reported to be expressed in the hypothalamus (35, 36). It is therefore possible that a yet uncharacterized central TSH–TSHR feedback circuit may directly regulate the hypothalamic–pituitary–thyroid axis, thought solely to be controlled by thyroid hormones.…”

Section: Discussionmentioning

confidence: 99%

Brain Atlas for Glycoprotein Hormone Receptors at Single-Transcript Level

Ryu

Гумерова

Korkmaz

et al. 2022

Preprint

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There is increasing evidence that anterior pituitary hormones, traditionally thought to have unitary functions in regulating single endocrine targets, act on multiple somatic tissues, such as bone, fat, and liver. There is also emerging evidence for anterior pituitary hormone action on brain receptors in mediating central neural and peripheral somatic functions. Here, we have created the most comprehensive neuroanatomical atlas on the expression of TSHRs, LHCGRs and FSHRs. We have used RNAscope, a technology that allows the detection of mRNA at single-transcript level, together with protein level validation, to document Tshr expression in 173 and Fshr expression in 353 brain regions, nuclei and sub–nuclei identified using the Atlas for the Mouse Brain in Stereotaxic Coordinates. We also identified Lhcgr transcripts in 401 brain regions, nuclei and sub–nuclei. Complementarily, we used ViewRNA, another single-transcript detection technology, to establish the expression of FSHRs in human brain samples, where transcripts were co–localized in MALAT1–positive neurons. In addition, we show high expression for all three receptors in the ventricular region—with yet unknown functions. Intriguingly, Tshr and Fshr expression in the ependymal layer of the third ventricle was similar to that of the thyroid follicular cells and testicular Sertoli cells, respectively. TSHRs were expressed specifically in tanycytes. In contrast, Fshrs were localized to NeuN–positive neurons in the granular layer of the dentate gyrus in murine and human brain—both are Alzheimer's disease vulnerable regions. Our atlas thus provides a vital resource for scientists to explore the link between the stimulation or inactivation of brain glycoprotein hormone receptors on somatic function. New actionable pathways for human disease may be unmasked through further studies.

show abstract

Subcellular Localization of Immunoreactive Thyroid-Stimulating Hormone in the Rat Hypothalamus

Cited by 10 publications

References 9 publications

Immunoreactive Prolactin in the Rat Hypothalamus: In vitro Release and Subcellular Localization

Immunoreactive Prolactin in the Rat Hypothalamus: In vitro Release and Subcellular Localization

Brain atlas for glycoprotein hormone receptors at single-transcript level

Brain Atlas for Glycoprotein Hormone Receptors at Single-Transcript Level

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