Physiological Characterization and Transcriptomic Properties of GnRH Neurons Derived From Human Stem Cells

Autor: Kim L. Keen, Andrew J. Petersen, Serkan Erdin, Jaeweon Shin, Robert A. Pearce, Anita Bhattacharyya, Alexander G Figueroa, Benjamin I. Fordyce, Michael E. Talkowski, Ei Terasawa, Rachita Yadav
Rok vydání: 2021
Předmět:
0301 basic medicine
induced pluripotent stem cell
endocrine system
medicine.medical_specialty
Fibroblast Growth Factor 8
Neurogenesis
Human Embryonic Stem Cells
Induced Pluripotent Stem Cells
Biology
kisspeptin
Gonadotropin-Releasing Hormone
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Endocrinology
FGF8
Kisspeptin
Internal medicine
medicine
Humans
Induced pluripotent stem cell
Research Articles
Cells
Cultured
Neurons
Gene Expression Profiling
Gene Expression Regulation
Developmental
Cell Differentiation
Embryonic stem cell
Neuroepithelial cell
030104 developmental biology
chemistry
Hypothalamus
GnRH
human stem cell
Stem cell
Neurokinin B
Transcriptome
AcademicSubjects/MED00250
hormones
hormone substitutes
and hormone antagonists
030217 neurology & neurosurgery
Zdroj: Endocrinology
ISSN: 1945-7170
0013-7227
DOI: 10.1210/endocr/bqab120
Popis: Gonadotropin-releasing hormone (GnRH) neurons in the hypothalamus play a key role in the regulation of reproductive function. In this study, we sought an efficient method for generating GnRH neurons from human embryonic and induced pluripotent stem cells (hESC and hiPSC, respectively). First, we found that exposure of primitive neuroepithelial cells, rather than neuroprogenitor cells, to fibroblast growth factor 8 (FGF8), was more effective in generating GnRH neurons. Second, addition of kisspeptin to FGF8 further increased the efficiency rates of GnRH neurogeneration. Third, we generated a fluorescent marker mCherry labeled human embryonic GnRH cell line (mCh-hESC) using a CRISPR-Cas9 targeting approach. Fourth, we examined physiological characteristics of GnRH (mCh-hESC) neurons: similar to GnRH neurons in vivo, they released the GnRH peptide in a pulsatile manner at ~60 min intervals; GnRH release increased in response to high potassium, kisspeptin, estradiol, and neurokinin B challenges; and injection of depolarizing current induced action potentials. Finally, we characterized developmental changes in transcriptomes of GnRH neurons using hESC, hiPSC, and mCh-hESC. The developmental pattern of transcriptomes was remarkably similar among the 3 cell lines. Collectively, human stem cell–derived GnRH neurons will be an important tool for establishing disease models to understand diseases, such as idiopathic hypothalamic hypogonadism, and testing contraceptive drugs.
Databáze: OpenAIRE
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