德晋贵宾厅

• POMC is a complex precursor protein that hydrolyzed to produce a variety of neuroendocrine peptides. Under the action of hypothalamic corticotropin releasing hormone (CRH), the anterior pituitary gland hydrolyzed and secreted ACTH in the basophile cells of the pituitary gland. ACTH was highly expressed in the pituitary gland and partially expressed in the hypothalamus, adrenal medulla, intestine and placenta. In addition, POMC is also hydrolyzed to α-MSH, β-MSH, and γ-MSH, including ACTH, which is the ligand of MC1R, MC3R, MC4R, and MC5R, and ACTH is the only ligand of MC2R. ACTH is an important hormone to maintain the normal form and function of adrenal gland, promote the synthesis and secretion of adrenal glucocorticoid (cortisol), and play a key role in regulating body metabolism and stress response.
• It can be used to evaluate the efficacy or safety of drugs associated with abnormal ACTH, such as Cushing syndrome, congenital adrenal hyperplasia and ectopic ACTH syndrome.

Gene targeting strategy for B-hPOMC mice. The exons 2~3 of mouse Pomc gene that encodes the full-length protein, including 3’UTR were replaced by human POMC exons 2~3 in B-hPOMC mice. The promoter and 5’UTR region of the mouse gene are retained. The human POMC expression is driven by endogenous mouse Pomc promoter, while mouse Pomc gene transcription and translation will be disrupted.

Strain specific analysis of POMC mRNA expression in wild-type C57BL/6N mice and B-hPOMC mice by RT-PCR. Testis RNA were isolated from wild-type C57BL/6N mice (+/+) and homozygous B-hPOMC mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human POMC primers. Mouse Pomc mRNA was only detectable in wild-type mice. Human POMC mRNA was exclusively detectable in homozygous B-hPOMC mice but not in wild-type mice.

Western blot analysis of POMC protein expression in homozygous B-hPOMC mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hPOMC mice (H/H), and then analyzed by western blot with anti-POMC antibody(abcam, ab254257). 40 μg total proteins were loaded for western blotting analysis. POMC was detected in Pituitary, cerebral cortex, testis.