Search

Nucleobindin-derived peptides, nesfatin-1 [NESF-1] and nesfatin-1-like-peptide [NLP] have diverse roles in endocrine and metabolic regulation. While both peptides showed a stimulatory effect on the synthesis of proopiomelanocortin (POMC), the adrenocorticotropic hormone (ACTH) precursor in mouse corticotrophs, whether NESF-1 and NLP have any direct effect on glucocorticoid [GC] synthesis in the adrenal cortex remains unknown. The main aim of this study was to determine if NESF-1 and/or NLP act directly on adrenal cortex cells to regulate cortisol synthesis in vitro. Whether NLP injection affects stress-hormone gene expression in the adrenal gland and pituitary in vivo in mice was also assessed. In addition, cortisol synthetic pathway in Nucb1 knockout mice was studied. Human adrenal cortical [H295R] cells showed immunoreactivity for both NUCB1/NLP and NUCB2/NESF-1. NLP and NESF-1 decreased the abundance of steroidogenic enzyme mRNAs, and cortisol synthesis and release through the AC/PKA/CREB pathway in H295R cells. Similarly, intraperitoneal injection of NLP in mice decreased the expression of enzymes involved in glucocorticoid (GC) synthesis in the adrenal gland while increasing the expression of Pomc, Pcsk1 and Crhr1 in the pituitary. Moreover, the melanocortin 2 receptor (Mc2r) mRNA level was enhanced in the adrenal gland samples of NLP injected mice. However, the global genetic disruption in Nucb1 did not affect most steroidogenic enzyme mRNAs, and Pomc, Pcsk2 and Crhr1 mRNAs in mice adrenal gland and pituitary gland, respectively. Collectively, these data provide the first evidence for a direct inhibition of cortisol synthesis and secretion by NLP and NESF-1. NUCB peptides might still elicit a net stimulatory effect on GC synthesis and secretion through their positive effects on ACTH-MC2R pathway in the pituitary. [ABSTRACT FROM AUTHOR]

Nucleobindin, a Ca2+-binding protein, has been previously identified within the nucleus and endoplasmic reticulum, and in association with the Golgi membrane. In addition, nucleobindin has been shown to be a minor constituent of bone extracellular matrix and has been postulated to play a role in mineralization. In the current investigation, we report the expression and localization of nucleobindin within odontoblasts and the dentin matrix. Nucleobindin mRNA transcripts were detected in the tooth, and in situ hybridization analysis substantiated the findings, showing nucleobindin expression within mature odontoblasts and within the cells of surrounding developing alveolar bone. Western blot analysis of tooth protein extracts demonstrated the presence of a 63 kDa protein, which showed immunologic affinity for a rat nucleobindin peptide antibody. The distribution of the protein was shown in mature odontoblasts by using immunohistochemistry. Moreover, immunogold labeling of nucleobindin and subsequent ultrastructural analysis demonstrated a similar pattern of distribution. Nucleobindin was identified within odontoblast cellular compartments: the nucleus, endoplasmic reticulum, and mitochondria. Of interest, nucleobindin localization was observed within the surrounding dentin extracellular matrix, and immunogold labeling was shown to accumulate with tissue development toward the cusp. The study clearly demonstrated the presence of nucleobindin within dental tissues. In consideration of the known functional properties of nucleobindin, it may be postulated that nucleobindin may contribute to the accumulation and transport of Ca2+ ions to the mineralization front prior to hydroxyapatite deposition. [ABSTRACT FROM AUTHOR]