Expression of follistatin mRNA by somatotropes and mammotropes early in the rat estrous cycle.

We previously found follistatin (FS) mRNA in gonadotropes [predominantly in cells with luteinizing hormone (LH) antigens] and folliculostellate cells (with S100 antigens) in diestrus rats pituitaries. However, earlier in the cycle, when percentages of gonadotropes are lowest, percentages of cells expressing FS are 1.5-2-fold higher than in diestrus. This study was designed to detect FS mRNA and other pituitary antigens to identify the additional cells with dual in situ hybridization and immunolabeling protocols. The mRNA was detected with biotinylated complementary oligonucleotide probes and avidin-biotin-peroxidase complexes. Significant labeling for FS mRNA was found in cells with the following antigens: growth hormone (GH) (7% of pituitary cells); prolactin (PRL) (5%); S100 protein (5%); follicle-stimulating hormone (FSH beta) (4%); LH beta (3%); and thyroid-stimulating hormone (TSH beta) (3%). Optimal conditions for detection included: overnight plating of > 50,000 cells/well (24-well tray) in media containing 10% fetal bovine serum; hybridization at 37 degrees C; and fixation in 2% glutaraldehyde. Whereas FS is expressed predominantly by LH gonadotropes at midcycle, FS mRNA can be expressed by all types of antigen-bearing cells earlier in the cycle. Its function in the pituitary may relate to its role in binding activin, which would result in inhibition of FSH release. However, since activin inhibits secretion of GH, PRL, and adrenocorticotropin (ACTH), FS may also control activin's effects on these cells. The FS-expressing cells may therefore be paracrine or autocrine regulators.

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Sensitivity of rat adenohypophyseal cells to estradiol and LHRH during long-term culture

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1981.240.6.e602 ◽

1981 ◽

Vol 240 (6) ◽

pp. E602-E608

Author(s):

L. Lagace ◽

F. Labrie ◽

T. Antakly ◽

G. Pelletier

Keyword(s):

Luteinizing Hormone ◽

Cell Types ◽

Thyroid Stimulating Hormone ◽

Time Interval ◽

Pituitary Cells ◽

Luteinizing Hormone Releasing Hormone ◽

Lh Release ◽

Lh Rh ◽

Stimulating Hormone

To determine possible effects of the time in culture on the responsiveness of the different pituitary cell types to estrogens, rat anterior pituitary cells were incubated up to 20 days in the presence or absence of 10 nM 17 beta-estradiol. Whereas spontaneous luteinizing hormone (LH) and thyroid-stimulating hormone (TSH) release decreased by 85-90%, follicle-stimulating hormone (FSH) and prolactin accumulation in medium were only 50% decreased after 20 days in culture, thus suggesting that the secretion of FSH and prolactin is less dependent on extrinsic stimulatory factors. Estradiol increased spontaneous LH release and its responsiveness to luteinizing hormone-releasing hormone (LH-RH) up to day 16 in culture, whereas the stimulatory effect of the estrogen on FSH secretion was significant only up to day 6. The stimulatory effect of estradiol on basal TSH release was seen up to day 8 in culture, whereas that on spontaneous prolactin release increased progressively after day 8 in culture up to the last time interval studied (20 days). As revealed by immunocytochemistry, the stimulatory effect of estradiol was not due to changes of cell growth.

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OR16-05 Single-Cell Sequencing Identifies Novel Regulators of Thyrotrope Populations and POU1F1-Independent Thyroid-Stimulating Hormone Expression

Journal of the Endocrine Society ◽

10.1210/jendso/bvaa046.655 ◽

2020 ◽

Vol 4 (Supplement_1) ◽

Author(s):

Leonard Cheung ◽

Alexandre Daly ◽

Michelle Brinkmeier ◽

Sally Ann Camper

Keyword(s):

Transcription Factor ◽

Single Cell ◽

Molecular Mechanisms ◽

Thyroid Stimulating Hormone ◽

Pars Tuberalis ◽

Pituitary Cells ◽

Pars Distalis ◽

Mouse Development ◽

Single Cell Sequencing ◽

Stimulating Hormone

Abstract We implemented single-cell RNA sequencing (scRNAseq) technology as a discovery tool to identify factors enriched in differentiated thyrotropes. Thyroid-stimulating hormone (TSH) is produced in the pars distalis of the anterior pituitary (AP) and primarily acts on the thyroid gland to regulate metabolism through T3/T4. However, TSH is also produced by cells in the pars tuberalis (PT), which is comprised of a thin layer of cells that extends rostrally from the pars distalis along the pituitary stalk to the median eminence in the hypothalamus. TSH produced by PT thyrotropes acts on hypothalamic tanycytes to regulate seasonal reproduction. PT thyrotropes likely descend from rostral tip thyrotropes that arise at e12.5 of mouse development, which transcribe the TSH beta subunit (Tshb) without detectable expression of the transcription factor POU1F1. POU1F1 is required for Tshb transcription in thyrotropes of the adenohypophysis, and it acts synergistically with GATA2 to drive cell fate. The molecular mechanisms driving Tshb expression independently of Pou1f1 in PT thyrotropes are unclear. Thyrotropes are the least abundant endocrine cell-type in the pituitary gland. We used genetic labeling and fluorescence-activated cell sorting (FACS) to enrich for thyrotropes for single-cell sequencing. We performed scRNAseq on 7-day-old GFP-positive pituitary cells from Tshb-Cre; R26-LSL-eYFP and intact whole pituitaries, recovering more than 15,000 cells altogether. We observe two distinct populations of cells expressing Tshb. The larger thyrotrope population has approximately twenty fold higher levels of Tshb and five fold higher Cga transcripts than the smaller population, and they are also distinguished by expression of Pou1f1, TSH-releasing hormone receptor (Trhr), and deiodinase 2 (Dio2), consistent with expectations for AP thyrotropes. The smaller thyrotrope population does not express Pou1f1, but those cells are characterized by expression of TSH receptor (Tshr) and melatonin receptor 1A (Mtnr1a), consistent with expectations for PT thyrotropes. They express mildly increased levels of Eya3 and Six1, although these genes are expressed in other cell-types including AP thyrotropes, stem cells, and gonadotropes. They have two-fold higher levels of Gata2 transcripts and uniquely express the transcription factor Sox14. SOX14 is a SoxB2 family transcription factor that counteracts the transcriptional activity of SoxB1 family members, such as Sox2. In conclusion, our scRNAseq has identified novel markers of PT thyrotropes and unveils novel insights into the similarities and differences in the development and function of pituitary thyrotrope subpopulations.

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Long-term adsorption of fetal bovine serum on H/O-terminated diamond studied in situ by atomic force microscopy

physica status solidi (b) ◽

10.1002/pssb.200982257 ◽

2009 ◽

Vol 246 (11-12) ◽

pp. 2832-2835 ◽

Cited By ~ 17

Author(s):

E. Ukraintsev ◽

B. Rezek ◽

A. Kromka ◽

A. Broz ◽

M. Kalbacova

Keyword(s):

Atomic Force Microscopy ◽

Bovine Serum ◽

Fetal Bovine Serum ◽

Force Microscopy ◽

Atomic Force ◽

Fetal Bovine

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THYROID-STIMULATING HORMONE AND GROWTH HORMONE RELEASE ALTERATIONS INDUCED BY MOSQUITO LARVAE PROTEINS ON PITUITARY CELLS

Cell Biology International ◽

10.1006/cbir.2001.0716 ◽

2001 ◽

Vol 25 (9) ◽

pp. 885-892 ◽

Cited By ~ 2

Author(s):

F Bolognani

Keyword(s):

Growth Hormone ◽

Thyroid Stimulating Hormone ◽

Mosquito Larvae ◽

Pituitary Cells ◽

Hormone Release ◽

Growth Hormone Release ◽

Stimulating Hormone

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EVALUASI OOSIT KAMBING HASIL IVM SEBAGAI SALAH SATU FAKTOR PENENTU KEBERHASILAN DALAM AKTIVASI PARTENOGENESIS

el–Hayah ◽

10.18860/elha.v2i1.1791 ◽

2012 ◽

Vol 2 (1) ◽

Author(s):

Kholifah Holil

Keyword(s):

Bovine Serum ◽

Fetal Bovine Serum ◽

Polar Body ◽

Follicle Stimulating Hormone ◽

Fetal Bovine ◽

Stimulating Hormone

Aktivasi partenogenesis merupakan salah satu tehnik aktivasi oosit untuk menghasilkan embrio tanpa kontribusi dari sperma. Salah satu faktor yang menentukan keberhasilan tehnik ini adalah pada ketersediaan oosit yang berkualitas. Oleh karena itu penelitian ini bertujuan untuk mengevaluasi oosit kambing hasil IVM yang dapat digunakan untuk kepentingan dalam aktivasi partenogenesis tersebut. Sampel yang digunakan dalam penelitian ini adalah oosit yang diaspirasi dari folikel ovarium kambing yang diambil dari RPH Sukun Malang. Oosit di IVM selama 24 jam dan selama 27 jam dalam medium TCM-199 yang ditambah dengan fetal bovine serum (FBS), follicle-stimulating hormone (FSH) dan lutheinizing hormone (LH) dan diinkubasi pada suhu 38,5oC, 5% CO2. Pada jam ke 24 dan jam ke 27 setelah IVM dilakukan pengamatan yang meliputi ekspansi sel-sel kumulus dan keberadaan polar body I (PB-I) pada jam ke 30. Berdasarkan hasil penelitian menunjukkan bahwa oosit kambing yang di IVM sampai jam ke 24 belum menunjukkan adanya ekspansi sel-sel kumulus kualitas 2 dan penampakan PB-I. Hasil tersebut berbeda dengan oosit yang di IVM sampai jam ke 27. Pada pengamatan jam ke 27 ini menunjukkan bahwa terdapat 77,87% oosit yang sel-sel kumulusnya berekspansi (kualitas 2) dan PB-I yang nampak sebesar 95,32%. Dengan demikian maka dapat disimpulkan bahwa oosit kambing hasil IVM baru dapat digunakan untuk keperluan lebih lanjut khususnya untuk keperluan aktivasi partenogenesis pada jam ke 27.Kata kunci: evaluasi, aktivasi partenogenesis, oosit kambing hasil IVM

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Application of a rapid avidin--biotin--peroxidase complex (ABC) technique to the localization of pituitary hormones at the electron microscopic level.

Journal of Histochemistry & Cytochemistry ◽

10.1177/30.12.6296223 ◽

1982 ◽

Vol 30 (12) ◽

pp. 1320-1324 ◽

Cited By ~ 29

Author(s):

G V Childs ◽

G Unabia

Keyword(s):

Staining Intensity ◽

Thyroid Stimulating Hormone ◽

Electron Microscopic Level ◽

Pituitary Cells ◽

Electron Microscopic ◽

High Background ◽

Prolonged Incubation ◽

Abc Method ◽

Rat Thyroid ◽

Stimulating Hormone

The new avidin--biotin--peroxidase complex (ABC) technique was applied to ultrathin sections of rat pituitary that were fixed with glutaraldehyde and embedded in Araldite 6005. The primary antisera dilutions that are normally applied for 24-48 hr with the peroxidase-antiperoxidase (PAP) complex technique were used. High background was observed with the ABC method when incubation times were 12-48 hr. Tests were then conducted with shorter incubation times. The staining intensity was measured with a densitometer. Detectable stain was seen after only 15 min in dilutions of 1:10,000 anti-bovine luteinizing hormone (bLH beta), 1:8000 anti-rat thyroid-stimulating hormone (rTSH beta), and 1:20,000 anti-25-39-adrenocorticotropic hormone (25-39ACTH). Optimal LH staining was seen after 30 min, whereas optimal staining for TSH or ACTH required 1 hr. Stain was detectable with a dilution of 1:4000 anti-human follicle-stimulating hormone (hFSH beta) after 30 min and was optimal after 4 hr. Prolonged incubation times with these dilutions decreased the staining intensity because a deposit of high background was produced that appeared as a filigreed network over the cells. When higher dilutions were tested with 2-hr incubation times, optimal staining was seen with 1:30,000 anti-bLH beta, 1:24,000 anti-rTSH beta, 1:30,000 anti-25-39ACTH, and 1:8000 anti-hFSH beta. These tests demonstrate the potential of the ABC method for the rapid detection of small amounts of specific and nonspecific antibodies that are bound to pituitary cells.

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Combined use of immunocytochemistry and in situ hybridization to study β thyroid-stimulating hormone gene expression in pituitaries of hypothyroid rats

Molecular and Cellular Probes ◽

10.1016/0890-8508(90)90029-y ◽

1990 ◽

Vol 4 (5) ◽

pp. 385-396 ◽

Cited By ~ 6

Author(s):

Jennifer H. Steel ◽

Domhnall J. O'Halloran ◽

Philip M. Jonesa ◽

Susan Van Noorden ◽

William W. Chin ◽

...

Keyword(s):

Gene Expression ◽

In Situ Hybridization ◽

Thyroid Stimulating Hormone ◽

Combined Use ◽

Stimulating Hormone

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Adsorption of Proteins Derived from Fetal Bovine Serum onto Hydroxyapatite Nanocrystals with Quartz Crystal Microbalance Technique

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.396-398.47 ◽

2008 ◽

Vol 396-398 ◽

pp. 47-50 ◽

Cited By ~ 7

Author(s):

Motohiro Tagaya ◽

Toshiyuki Ikoma ◽

Taro Takemura ◽

Mitsuhiro Okuda ◽

Nobutaka Hanagata ◽

...

Keyword(s):

Quartz Crystal Microbalance ◽

Bovine Serum ◽

Quartz Crystal ◽

Fetal Bovine Serum ◽

Protein Films ◽

Carbonate Buffer ◽

Adsorption Behaviors ◽

Fetal Bovine ◽

Hydroxyapatite Nanocrystals

The adsorption of multiple proteins derived from fetal bovine serum (FBS) in phosphate buffer saline (PBS) and alpha minimum essential (aMEM) was in situ analyzed with a quartz crystal microbalance with dissipation technique on gold, titanium and HAp sensors. The adsorption behaviors of FBS proteins were varied depending on the sensors. The DD/Df value of the HAp sensor were clearly different in PBS and aMEM, and others were not changed. The viscoelastic properties of the protein films adsorbed on the HAp sensor in PBS were flexible in comparison with those on the gold and titanium sensors. The D-f plots incidated that the proteins adsorbed on HAp in PBS would lead to competitive adsorption and conformational change and those in aMEM could form a monolayer. The adsorption behavior on the HAp in carbonate buffer saline was found to be similar to that in aMEM. These differential adsorption behaviors on the HAp surface were attributed to the pre-adsorptive ion, such PO43- or CO32- in the solvent.

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